Patent Application
20230127830
In recent years, viral and bacterial infection is becoming more prevalent worldwide and presents a serious public health threat. For example, the Coronavirus-2019 (COVID-19) global pandemic of a respiratory disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has affected over 435 million people worldwide, including nearly 6 million deaths, and is exacerbated by a lack of officially approved therapeutics as well as a lack of thoroughly tested, proven safe and effective vaccines. Several promising therapeutic agents are currently undergoing active investigation and development for prophylactic or therapeutic use in the treatment for COVID-19 to prevent or ameliorate its damaging effects to the afflicted patients, while in the meantime experimental vaccines are widely distributed to the general population.
Accordingly, there exists an urgent need for new and meaningful methods to supplement therapeutic and vaccination efforts by way of preventing or treating symptoms of long COVID or post-acute COVID syndrome (PACS) among patients who are recovering from the disease, including adults and children, to achieve reduction or elimination of long lingering clinical symptoms and their associated effects. The purpose of this study is to identify gut microbial species that can potentially provide the beneficial effects of preventing or treating PACS as well as to identify gut microbial species that can be monitored among patients suffering from active COVID to indicate individuals who are likely to develop PACS symptoms in the future, who can then be given treatment accordingly in a prophylactic manner. Direct supplementation of these beneficial gut microbial species is a potentially effective means to achieve these goals in the efforts of reducing or eliminating the long-term ill effects of infectious diseases such as COVID-19. The present invention fulfills this and other related needs by identifying beneficial gut microorganisms so as to allow formulation of new compositions and application of new methods that are effective for addressing lingering clinical symptoms among recovered COVID patients, as well as by identifying gut microorganisms that can serve as markers to indicate COVID patients who have an increased risk of developing PACS as to allow these patients to be treated prophylactically to prevent the onset of long COVID symptoms.
The present inventors discovered in their studies the certain gut microbial species and their metabolites can help prevent and treat the symptoms of long COVID or post-acute COVID syndrome (PACS) among patients who have been diagnosed of COVID-19, including those having recovered from the disease (e.g., having had at least one negative test report for SARS-CoV2, the virus causing the infection), thus provide an important utility in facilitating patients' complete recovery from the disease and elimination of potential lingering ill effects caused by this pathogen. The microorganisms so identified now serve to provide new methods and compositions as an integral part of the COVID-19 therapy and long-term management.
In a first aspect, the present invention provides a composition that is useful for use in treating PACS in a subject comprising an effective amount of (1) one or more beneficial bacterial species selected from Table 1 and Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum; and (2) a physiologically acceptable excipient. In some embodiments, the composition further comprises one or more of beneficial bacterial species selected from Table 3. In some embodiments, the composition comprises a total of about 106 to about 1012 colony-forming unit (CFU) of the beneficial bacterial species. In some embodiments, the composition comprises no detectable amount of any bacterial species in Table 4 including Scardovia wiggsiae and Bacteroides xylanisolvens. In some embodiments, the composition comprises an effective amount of one or more different beneficial bacterial species of Bifidobacterium longum, Blautia wexlerae, Bifidobacterium pseudocatenulatum, Faecalibacterium prausnitzii, and Fusicatenibacter saccharivorans. In some embodiments, the composition comprises an effective amount of one or more different butyrate-producing bacterial species of Roseburia inulinivorans, Faecalibacterium prausnitzii, and Roseburia Hominis. In some embodiments, the composition consists essentially of an effective amount of (1) one or more of the beneficial bacterial species selected from Table 1 and Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum; (2) one or more physiologically acceptable excipients. In some embodiments, the composition is formulated for oral ingestion, such as in the form of a food or beverage item. In some embodiments, the composition is formulated for direct deposit to the subject's gastrointestinal tract. Optionally, the composition may further includes one or more agents known to enhance immunity, for example, effective in suppressing an infectious agent, such as SARS-CoV-2, and reducing disease severity (e.g., ivermectin, hydroxychloroquine, Zinc, vitamin C, vitamin D, quercetin, melatonin, fluvoxamine or fluoxetine, proxalutamide, azithromycin, doxycycline, or any combination thereof).
In the second aspect, the present invention provides a method for treating or preventing PACS in a subject by administering to the subject an effective amount of the composition described above and herein. In some embodiments, the subject has already developed PACS. In some embodiments, the subject has been diagnosed of COVID but has not yet developed PACS. In some embodiments, the administering step comprises administering to the subject one composition comprising one or more of the beneficial bacterial species. In some embodiments, the administering step comprises administering to the subject two or more compositions, each comprising one or more of the beneficial bacterial species. In some embodiments, the administering step comprises oral ingestion of the composition(s), which may be prior to or with food intake, or shortly after food intake (e.g., a meal or snack). In some embodiments, the administering step comprises direct deposit to the subject's gastrointestinal tract. For example, the technique knowns as fetal microbiota transplant (FMT) is used wherein a composition made from processed suitable donor fecal material containing the desirable amount of the beneficial bacterial species may be used to introduce the bacteria into a COVID or PACS patient's gastrointestinal tract. Optionally, the method also includes administering to the subject, either in the same composition(s) containing the beneficial bacteria, may further includes one or more agents known to enhance immunity, for example, effective in suppressing an infectious agent, such as SARS-CoV-2, and reducing disease severity (e.g., ivermectin, hydroxychloroquine, Zinc, vitamin C, vitamin D, quercetin, melatonin, fluvoxamine or fluoxetine, proxalutamide, azithromycin, doxycycline, or any combination thereof).
In a related aspect, the present invention provides a novel use of a composition in treating or preventing PACS in a subject. The composition comprising an effective amount of (1) one or more beneficial bacterial species selected from Table 1 and Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum; and (2) a physiologically acceptable excipient. In some embodiments, the composition further comprises one or more of beneficial bacterial species selected from Table 3. In some embodiments, the composition comprises a total of about 106 to about 1012 colony-forming unit (CFU) of the beneficial bacterial species. In some embodiments, the composition comprises no detectable amount of any bacterial species in Table 4 including Scardovia wiggsiae and Bacteroides xylanisolvens In some embodiments, the composition comprises an effective amount of one or more different beneficial bacterial species of Bifidobacterium longum, Blautia wexlerae, Bifidobacterium pseudocatenulatum, Faecalibacterium prausnitzii, and Fusicatenibacter saccharivorans. In some embodiments, the composition comprises an effective amount of one or more different butyrate-producing bacterial species of Roseburia inulinivorans, Faecalibacterium prausnitzii, and Roseburia Hominis. In some embodiments, the composition consists essentially of an effective amount of (1) one or more of the beneficial bacterial species selected from Table 1 and Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum; (2) one or more physiologically acceptable excipients. In some embodiments, the composition is formulated for oral ingestion, such as in the form of a food or beverage item. In some embodiments, the composition is formulated for direct deposit to the subject's gastrointestinal tract. Optionally, the composition may further includes one or more agents known to enhance immunity, for example, effective in suppressing an infectious agent, such as SARS-CoV-2, and reducing disease severity (e.g., ivermectin, hydroxychloroquine, Zinc, vitamin C, vitamin D, quercetin, melatonin, fluvoxamine or fluoxetine, proxalutamide, azithromycin, doxycycline, or any combination thereof).
In a third aspect, the present invention provides a kit for treating or preventing PACS in a subject. The kit includes multiple containers, each containing a distinct composition comprising an effective amount of one or more beneficial bacterial species selected from Table 1 and Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum. In some embodiments, the kit comprises a plurality of containers, each containing a distinct composition comprising an effective amount of one or more different beneficial bacterial species selected from Bifidobacterium longum, Blautia wexlerae, Bifidobacterium pseudocatenulatum, Faecalibacterium prausnitzii, and Fusicatenibacter saccharivorans. In some embodiments, the kit includes a plurality of containers, each containing a distinct composition comprising an effective amount of one or more different butyrate-producing bacterial species of Roseburia inulinivorans, Faecalibacterium prausnitzii, and Roseburia Hominis. In some embodiments, the compositions are in the form of a powder, liquid, paste, cream, tablet, or capsule. The kit may further include one or more additional containers, each containing one or more agents known to effectively suppress an infectious agent such as SARS-CoV-2 and/or to reduce disease severity or facilitate patient recovery from COVID-19 (e.g., ivermectin, vitamin C, vitamin D, quercetin, melatonin, Zinc, azithromycin, doxycycline, hydroxychloroquine, fluvoxamine or fluoxetin, proxalutamide, or any combination thereof). Moreover, a user instruction manual may be included in the kit for the proper use of the kit, providing information such as suitable subjects for treatment and time, dosing etc.
In a fourth aspect, the present invention provides a method for assessing risk of PACS in a subject. The method includes these steps: (1) determining, in a stool sample from the subject, the level or relative abundance of one or more of the bacterial species set forth in Table 2; (2) determining the level or relative abundance of the same bacterial species in a stool sample obtained at the time of initial diagnosis of COVID-19 (such as at the time of first positive test report for COVID-19 or at the time of admission to a hospital) from a reference cohort comprising COVID-19 patients who would or would not eventually develop PACS; (3) generating decision trees by random forest model using data obtained from step (2) and running the level or relative abundance of one or more of the bacterial species from step (1) down the decision trees to generate a score; and (4) determining the subject with a score greater than 0.5 as having an increased risk for PACS, and determining the subject with a score no greater than 0.5 as having no increased risk for PACS. In some embodiments, each of steps (1) and (2) comprises metagenomics sequencing. In some embodiments, each of steps (1) and (2) comprises a polymerase chain reaction (PCR), e.g., a quantitative PCR (qPCR). In some embodiments, the bacterial species comprise one or more of Bifidobacterium longum, Blautia wexlerae, Bifidobacterium pseudocatenulatum, Faecalibacterium prausnitzii, and Fusicatenibacter saccharivorans.
As used herein, the term “SARS-CoV-2 or severe acute respiratory syndrome coronavirus 2,” refers to the virus that causes Coronavirus Disease 2019 (COVID-19). It is also referred to as the “COVID-19 virus.”
The term “post-acute COVID-19 syndrome (PACS)” or “long COVID” is used to describe a medical condition in which a patient who has recovered from COVID, as indicated by a negative PCR report at least 2 weeks prior (e.g., from at least 3 or 4 weeks earlier), yet continuously and stably exhibits one or more symptoms of the disease without any notable progression. The symptoms may include respiratory (cough, sputum, nasal congestion/runny nose, shortness of breath), neuropsychiatric (headache, dizziness, loss of taste, loss of smell, anxiety, difficulty in concentration, difficulty in sleeping, sadness, poor memory, blurred vision), gastrointestinal (nausea, diarrhoea, abdominal pain, epigastric pain), dermatological (hair loss), or musculoskeletal (joint pain, muscle pain) symptoms, as well as fatigue.
The term “inhibiting” or “inhibition,” as used herein, refers to any detectable negative effect on a target biological process, such as RNA/protein expression of a target gene, the biological activity of a target protein, cellular signal transduction, cell proliferation, presence/level of an organism especially a micro-organism, any measurable biomarker, bio-parameter, or symptom in a subject, and the like. Typically, an inhibition is reflected in a decrease of at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater in the target process (e.g., a subject's bodyweight, or the blood glucose/cholesterol level, or any measurable symptom or biomarker in a subject, such as an infection rate among subjects by a pathogenic infectious agent), or any one of the downstream parameters mentioned above, when compared to a control. “Inhibition” further includes a 100% reduction, i.e., a complete elimination, prevention, or abolition of a target biological process or signal. The other relative terms such as “suppressing,” “suppression,” “reducing,” and “reduction” are used in a similar fashion in this disclosure to refer to decreases to different levels (e.g., at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater decrease compared to a control level) up to complete elimination of a target biological process or signal. On the other hand, terms such as “activate,” “activating,” “activation,” “increase,” “increasing,” “promote,” “promoting,” “enhance,” “enhancing,” or “enhancement” are used in this disclosure to encompass positive changes at different levels (e.g., at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, or greater such as 3, 5, 8, 10, 20-fold increase compared to a control level in a target process, signal, or parameter.
As used herein, the term “treatment” or “treating” includes both therapeutic and preventative measures taken to address the presence of a disease or condition or the risk of developing such disease or condition at a later time. It encompasses therapeutic or preventive measures for alleviating ongoing symptoms, inhibiting or slowing disease progression, delaying of onset of symptoms, or eliminating or reducing side-effects caused by such disease or condition. A preventive measure in this context and its variations do not require 100% elimination of the occurrence of an event; rather, they refer to a suppression or reduction in the likelihood or severity of such occurrence or a delay in such occurrence.
The term “severity” of a disease refers to the level and extent to which a disease progresses to cause detrimental effects on the well-being and health of a patient suffering from the disease, such as short-term and long-term physical, mental, and psychological disability, up to and including death of the patient. Severity of a disease can be reflected in the nature and quantity of the necessary therapeutic and maintenance measures, the time duration required for patient recovery, the extent of possible recovery, the percentage of patient full recovery, the percentage of patients in need of long-term care, and mortality rate.
A “patient” or “subject” receiving the composition or treatment method of this invention is a human, including both adult and juvenile human, of any age, gender, and ethnic background, who has been diagnosed with COVID-19 (e.g., has had a positive nucleic acid and/or antibody test result for SARS-CoV2) and is in need of being treated to address PACS symptoms or to prevent the onset of such symptoms. Typically, the patient or subject receiving treatment according to the method of this invention to prevent or treat long COVID symptoms is not otherwise in need of treatment by the same therapeutic agents. For example, if a subject is receiving the symbiotic composition according to the claimed method, the subject is not suffering from any disease that is known to be treated by the same therapeutic agents. Although a patient may be of any age, in some cases the patient is at least 20, 30, 40, 45, 50, 55, 60, 65, 70, 75, 80, or 85 years of age; in some cases, a patient may be between 20 and 30, 30 and 40, 40 and 45 years old, or between 50 and 65 years of age, or between 65 and 85 years of age. A “child” subject is one under the age of 18 years, e.g., about 5-17, 9 or 10-17, or 12-17 years old, including an “infant,” who is younger than about 12 months old, e.g., younger than about 10, 8, 6, 4, or 2 months old, whereas an “adult” subject is one who is 18 years or older.
The term “effective amount,” as used herein, refers to an amount that produces intended (e.g., therapeutic or prophylactic) effects for which a substance is administered. The effects include the prevention, correction, or inhibition of progression of the symptoms of a particular disease/condition and related complications to any detectable extent, e.g., incidence of disease, infection rate, one or more of the symptoms of a viral or bacterial infection and related disorder (e.g., COVID-19). The exact amount will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); and Pickar, Dosage Calculations (1999)).
The term “about” when used in reference to a given value denotes a range encompassing ±10% of the value.
A “pharmaceutically acceptable” or “pharmacologically acceptable” excipient is a substance that is not biologically harmful or otherwise undesirable, i.e., the excipient may be administered to an individual along with a bioactive agent without causing any undesirable biological effects. Neither would the excipient interact in a deleterious manner with any of the components of the composition in which it is contained.
The term “excipient” refers to any essentially accessory substance that may be present in the finished dosage form of the composition of this invention. For example, the term “excipient” includes vehicles, binders, disintegrants, fillers (diluents), lubricants, glidants (flow enhancers), compression aids, colors, sweeteners, preservatives, suspending/dispersing agents, film formers/coatings, flavors and printing inks.
The term “consisting essentially of,” when used in the context of describing a composition containing an active ingredient or multiple active ingredients, refer to the fact that the composition does not contain other ingredients possessing any similar or relevant biological activity of the active ingredient(s) or capable of enhancing or suppressing the activity, whereas one or more inactive ingredients such as physiological or pharmaceutically acceptable excipients may be present in the composition. For example, a composition consisting essentially of active agents (for instance, one or more bacterial species in Table 1) effective for treating PACS in a subject is a composition that does not contain any other agents that may have any detectable positive or negative effect on the same target process (e.g., any one of the PACS symptoms) or that may increase or decrease to any measurable extent of the relevant symptoms among the receiving subjects.
This invention describes specific bacterial species and combination thereof (e.g., beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum) for preventing or treating PACS symptoms in a subject, especially when the subject is a human adult or child suffering but recovering from COVID-19. The practical use of the invention includes development and manufacturing of commercial food products or health supplements, for example in the form of a powder, tablet, capsule, or liquid, which can be taken alone or added to food or beverages, as well as any other formulation suitable for use by fecal microbiota transplant (FMT).
The present invention provides pharmaceutical compositions comprising an effective amount of one or more of the beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum for treating a COVID-19 patient to reduce the risk of developing symptom(s) of PACS or to ameliorate the symptom(s) if any already present. Pharmaceutical compositions of the invention are suitable for use in a variety of drug delivery systems. Suitable formulations for use in the present invention are found in Remington's Pharmaceutical Sciences, Mack Publishing Company, Philadelphia, Pa., 17th ed. (1985). For a brief review of methods for drug delivery, see, Langer, Science 249: 1527-1533 (1990).
The pharmaceutical compositions of the present invention can be administered by various routes, e.g., systemic administration via oral ingestion or local delivery using a rectal suppository. The preferred route of administering the pharmaceutical compositions is oral administration at daily doses of about 106 to about 1012 CFU for the combination of all beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum. When multiple bacterial species are administered to the subject, they may be administered either in one single composition or in multiple compositions. The appropriate dose may be administered in a single daily dose or as divided doses presented at appropriate intervals, for example as two, three, four, or more subdoses per day. The duration of administration may range from about 1 week to about 8 weeks, e.g., about 2 week to about 4 weeks, or for a longer time period (e.g., up to 6 months) as the relevant symptoms persist.
For preparing pharmaceutical compositions containing the beneficial bacteria identified in this disclosure, one or more inert and pharmaceutically acceptable carriers are used. The pharmaceutical carrier can be either solid or liquid. Solid form preparations include, for example, powders, tablets, dispersible granules, capsules, cachets, and suppositories. A solid carrier can be one or more substances that can also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, or tablet disintegrating agents; it can also be an encapsulating material.
In powders, the carrier is generally a finely divided solid that is in a mixture with the finely divided active component, e.g., any one or more of the beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum. In tablets, the active ingredient is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.
For preparing pharmaceutical compositions in the form of suppositories, a low-melting wax such as a mixture of fatty acid glycerides and cocoa butter is first melted and the active ingredient is dispersed therein by, for example, stirring. The molten homogeneous mixture is then poured into convenient-sized molds and allowed to cool and solidify.
Powders and tablets preferably contain between about 5% to about 100% by weight of the active ingredient(s) (e.g., one or more of the beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum). Suitable carriers include, for example, magnesium carbonate, magnesium stearate, talc, lactose, sugar, pectin, dextrin, starch, tragacanth, methyl cellulose, sodium carboxymethyl cellulose, a low-melting wax, cocoa butter, and the like.
The pharmaceutical compositions can include the formulation of the active ingredient(s), e.g., one or more of the beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum, with encapsulating material as a carrier providing a capsule in which the active ingredient(s) (with or without other carriers) is surrounded by the carrier, such that the carrier is thus in association with the active ingredient(s). In a similar manner, sachets can also be included. Tablets, powders, sachets, and capsules can be used as solid dosage forms suitable for oral administration.
Liquid pharmaceutical compositions include, for example, solutions suitable for oral administration or local delivery, suspensions, and emulsions suitable for oral administration. Sterile water solutions of the active component (e.g., one or more of the beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum) or sterile solutions of the active component in solvents comprising water, buffered water, saline, PBS, ethanol, or propylene glycol are examples of liquid or semi-liquid compositions suitable for oral administration or local delivery such as by rectal suppository. The compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions, such as pH adjusting and buffering agents, tonicity adjusting agents, wetting agents, detergents, and the like.
Sterile solutions can be prepared by dissolving the active component (e.g., one or more of the beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum) in the desired solvent system, and then passing the resulting solution through a membrane filter to sterilize it or, alternatively, by dissolving the sterile active component in a previously sterilized solvent under sterile conditions. The resulting aqueous solutions may be packaged for use as is, or lyophilized, the lyophilized preparation being combined with a sterile aqueous carrier prior to administration. The pH of the preparations typically will be between 3 and 11, more preferably from 5 to 9, and most preferably from 7 to 8.
Single or multiple administrations of the compositions can be carried out with dose levels and pattern being selected by the treating physician. In any event, the pharmaceutical formulations should provide a quantity of an active agent sufficient to effectively enhance the efficacy of a vaccine and/or reduce or eliminate undesirable adverse effects of a vaccine.
Different combinations of bacteria in the pharmaceutical compositions can be chosen according to Table 1 to target specific categories of symptoms. For example, Agathobaculum butyriciproducens, Bifidobacterium pseudocatenulatum, Roseburia hominis, Anaerostipes hadrus, Dialister sp. CAG: 357, Eubacterium rectale can be used to prevent or alleviate fatigue; Agathobaculum butyriciproducens, Bifidobacterium pseudocatenulatum, Roseburia hominis, Asaccharobacter celatus can be used to prevent or alleviate gastrointestinal symptoms; Agathobaculum butyriciproducens, Roseburia hominis, Anaerostipes hadrus, Coprococcus comes can be used to prevent or alleviate neurological symptoms; Agathobaculum butyriciproducens, Bifidobacterium pseudocatenulatum, Roseburia hominis, Anaerostipes hadrus, Alistipes putredinis can be used to prevent or alleviate respiratory symptoms; Agathobaculum butyriciproducens, Anaerostipes hadrus, Eubacterium sp. CAG: 274, Dorea formicigenerans can be used to prevent or alleviate musculoskeletal symptoms; and Agathobaculum butyriciproducens, Roseburia hominis, Eubacterium rectale, Gemmiger formicilis, Eubacterium ventriosum, Dorea formicigenerans can be used to prevent or alleviate hair loss.
Additional known therapeutic agent or agents may be used in combination with an active agent such as one or more of the beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum in the practice of the present invention for the purpose of treating or preventing PACS symptom(s) in a patient recovering from COVID-19. In such applications, one or more of the previously known effective prophylactic/therapeutic agents can be administered to patients concurrently with an effective amount of the active agent(s) either together in a single composition or separately in two or more different compositions.
For example, drugs and supplements that are known to be effective for use to prevent or treat COVID-19 include ivermectin, vitamin C, vitamin D, melatonin, quercetin, Zinc, hydroxychloroquine, fluvoxamine/fluoxetine, proxalutamide, doxycycline, and azithromycin. They may be used in combination with the active agents (such as any one or more of the beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum) of the present invention to promote safe and full recovery among patients suffering from SARS-CoV2 infection, reduce potential disease severity (including morbidity and mortality), and ensure elimination of any lingering long-term ill effects from the disease. In particular, the combination of Zinc, hydroxychloroquine, and azithromycin and the combination of ivermectin, fluvoxamine or fluoxetine, proxalutamide, doxycycline, vitamin C, vitamin D, melatonin, quercetin, and Zinc have demonstrated high efficacy in both COVID prophylaxis and therapy. Thus, these known drug/supplement or nutritheutical combinations can be used in the method of this invention along with the active components of one or more of the beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum.
The invention also provides kits for treating and preventing PACS symptoms among patients recovering from COVID-19 in accordance with the method disclosed herein. The kits typically include a plurality of containers, each containing a composition comprising one or more of the beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum. Further, additional agents or drugs that are known to be therapeutically effective for prevention and/or treatment of the disease, including for ameliorating the symptoms and reducing the severity of the disease, as well as for facilitating recovery from the disease (such as those described in the last section or otherwise known in the pertinent technical field) may be included in the kit. The plurality of containers of the kit each may contain a different active agent/drug or a distinct combination of two or more of the active agents or drugs. The kit may further include informational material providing instructions on how to dispense the pharmaceutical composition(s), including description of the type of patients who may be treated (e.g., human patients, adults or children, who have been diagnosed of COVID-19 and now are in the process of recovering and are therefore seeking to eliminate lingering symptoms of the disease or to reduce the risk of such symptoms becoming persistent and continuous), the dosage, frequency, and manner of administration, and the like.
The present inventors also discovered that the altered level of certain bacterial species can indicate the prospect or likelihood of a COVID patient later develop PACS symptoms and therefore can provide a basis for prophylactic treatment: they revealed that the level or relative abundance of certain bacterial species (such as one or more of the species shown in Table 2) can be analyze to indicate an individual patient's risk or likelihood for later developing PACS symptoms when properly calculated using certain specified mathematic tools.
For example, when a stool sample taken from a COVID patient who is being tested is analyzed, the level or relative abundance of bacterial species in Table 2 in the sample is determined, e.g., by PCR especially quantitative PCR. In the meantime, the level or relative abundance of the same bacterial species is determined by the same method as the bacterial species is/are present in stool samples taken from individual COVID patients at the time of initial diagnosis of COVID-19 from a reference cohort comprising these COVID-19 patients, some of whom would eventually develop PACS whereas others would not develop PACS. Decision trees are then generated by random forest model using data obtained from the reference cohort, and the level or relative abundance of one or more of the bacterial species from the individual patient being tested is run down the decision trees to generate a score. The patient is deemed to have an increased risk for developing PACS when his score is greater than 0.5. In contrast, when his score is no greater than 0.5, the patient is deemed to have no increased risk for PACS.
Once the PACS risk assessment is made, for example, an individual who has been diagnosed as suffering from COVID-19 (e.g., based on a positive PCR or antibody test for SARS-CoV2) and who has been in the process of recovering from the disease is deemed to have an increased risk of developing PACS at a later time, appropriate treatment steps can be taken as a measure to achieve the goal of preventing the onset of the PACS symptoms or reducing their severity or eliminating them altogether. For instance, the patient may be given composition(s) comprising an effective amount of one or more of the beneficial bacterial species listed in Table 1 and Table 3 plus Bifidobacterium longum, Blautia wexlerae, and Bifidobacterium pseudocatenulatum, e.g., by FMT or by an alternative administration method via oral or local delivery, such that the bacterial profile in the patient's gastrointestinal tract will be modified to one that is favorable for the outcome of prevented, reduced, eliminated, or reversed PACS symptoms.
The following examples are provided by way of illustration only and not by way of limitation. Those of skill in the art will readily recognize a variety of non-critical parameters that could be changed or modified to yield essentially the same or similar results.
Clinical characteristics of coronavirus 2019 (COVID-19) during the acute infection is well described but little is known of long-term complications of COVID-19. Post-acute COVID-19 syndrome (PACS) characterised by long term complications and/or persistent symptoms beyond four weeks after the onset of COVID-19 is increasingly being recognised1-3. Up to three quarters of patients described at least one symptom at six months after recovery and multi-system symptoms including fatigue, muscle weakness and sleep difficulties are commonly reported4.
Reasons underlying the development of PACS is largely unclear. Perturbations of immune and inflammatory responses, cellular damage by acute viral infection or sequele of post critical illness may contribute to long term symptoms after COVID-19 infection5. As the gastrointestinal tract is the largest immunological organ in the body, aberrant immune response to COVID-19 infection induced by resident microorganisms may affect the recovery process. Emerging evidence supports the potential role of gut dysbiosis in severity of COVID-19 infection6. The present inventors herein investigated whether gut microbiota composition and to what extent affects risk of persistent symptoms in recovered patients with different severity of COVID-19 infection.
Gut microbiome composition and association with different symptoms were examined at six months. Based on permutational multivariate analysis of variance (PERMANOVA), gut composition was not associated with medical therapy during hospitalisation or disease severity (
A total of 81 bacterial species were associated with different categories of PACS and many of the bacteria species were associated with more than two categories of persistent symptoms (
It was discovered that very few patients were admitted to an intensive care unit or required mechanical ventilation, yet many developed PACS suggesting that persistent symptoms are unlikely to be the result of post critical illness syndrome. Loss of several symbionts including the genera Bifidobacteria, Roseburia, and Faecalibacteria known to have immunomodulatory functions were especially associated with persistent symptoms among recovered COVID-19 patients. The latter two bacteria are important short-chain acid (SCFAs) producers and major players in maintenance of immune homeostasis. SCFAs have been shown to alter chemotaxis and phagocytosis, induce reactive oxygen species, change cell proliferation and function, and have antimicrobial and anti-inflammatory effects.
A method is therefore provided for preventing and alleviating these symptoms by supplementation of these beneficial bacterial listed in Table 1 and Table 3. As these symptoms can also occur in the any persons including people with no history of COVID-19, such a method can be applied to anyone concerning these symptoms.
Species listed in Table 2, and preferably Bifidobacterium longum (NCBI:txid216816), Blautia wexlerae (NCBI:txid418240), Bifidobacterium pseudocatenulatum (NCBI:txid28026), Faecalibacterium prausnitzii (NCBI:txid853), Fusicatenibacter saccharivorans (NCBI:txid1150298), can be used in different combinations to build a risk assessment model to determine whether a person is at risk of PACS of COVID-19 after the person was tested positive for COVID-19.
To determine the risk of PACS in a subject, the following steps will be carried out:
The 6-minute walk test (6MWT) is frequently used to determine functional capacity in patients4. Previous work has shown that post-convalescence patients with lower microbiota richness had impaired lung function6, however, the cause of reduced 6-minute walk distance after COVID-19 remains unknown. Of the 68 COVID-19 patients who had provided stool samples at six-month follow up, 52 of them had 6-minute walk test assessment at six months. The median walking distance at 6 minutes in patients with PACS was significantly lower than those without PACS (mean 464 meter vs 382 meter, P<0.001,
Significant inverse associations of walking distance with pathogenic bacteria species (for example, Clostridium innocuum, Clostridium bolteae) were also observed, which could confer pathogenicity or were associated with disease risk in different populations10,11 (P<0.05,
Amongst gut bacteria species detected in patients with PACS, 28 bacteria species were diminished and 14 were enriched at baseline and follow-up samples (
Bacteria species such as Christensenella minuta, Eubacterium ramulus, Bifidobacterium adolescentis were found to be depleted at baseline sample of patients who developed PACS at six months (P<0.05, LefSe >2,
A composition comprising one or more beneficial bacteria selected from Table 3 or
This prospective cohort study was performed at three regional hospitals (Prince of Wales Hospital, United Christian Hospital and Yan Chai Hospital) in Hong Kong, China. All patients with a confirmed diagnosis of COVID-19, as evidenced by a positive SARS-CoV-2 by reverse transcriptase polymerase chain reaction test in nasopharyngeal swab, nasal swab, deep throat saliva, sputum, tracheal aspirate or stool, were recruited from 1 February to 31 Aug. 2020. All patients with confirmed COVID-19 were required to be hospitalised according to local government policy. Patients could be discharged if they fit either one of the below criteria: two clinical specimens of the same type (i.e., respiratory or stool) tested negative for nucleic acid of SARS-CoV-2 by RT-PCR taken at least 24 hours apart or tested positive for SARS-CoV-2 antibody. Patients were excluded if they were unable to be contacted, declined to participate in study or died before the follow-up visit. Data including demographics, clinical and laboratory data were extracted from electronic medical records in the Clinical Management System of the Hong Kong Hospital Authority. Severity of COVID-19 infection was categorized as (1) mild, if there was no radiographic evidence of pneumonia; (2) moderate, if pneumonia was present along with fever and respiratory tract symptoms; (3) severe, if respiratory rate ≥30/min, oxygen saturation ≤93% when breathing ambient air, or PaO2/FiO2≤300 mm Hg (1 mm Hg=0.133 kPa); or (4) critical, if there was respiratory failure requiring mechanical ventilation, shock, or organ failure requiring intensive care.12 Stool samples were collected serially from admission, at one month and at six months after discharge from hospital. Stool samples from in-hospital patients were collected by hospital staff while discharged patients provided stools on day of follow-up at 1 month and 6 months after discharge or self-sampled at home and couriered to the hospital within 24 hours of collection. All samples were collected in collection tubes containing preservative media (cat. 63700, Norgen Biotek Corp, Ontario Canada) and stored immediately at −80° C. until processing. It was previously shown that data of gut microbiota composition generated from stools collected using this preservative medium were comparable with data obtained from samples that were immediately stored at at −80° C.13. The presence of 30 most commonly reported symptoms post-COVID14,15 was assessed at three and six months after illness onset (Table 19). Post-acute COVID-19 syndrome (PACS) was defined as at least one persistent symptom which cannot be explained by alternative diagnosis four weeks after clearance of SARS-CoV-2. Six-minute walk distance test (6MWT), which is a simple functional assessment to assess the aerobic capacity and endurance, was performed at six months after discharge in a subset of recovered COVID-19 patient. The 6MWT distance data were correlated with the gut microbiota analysis. Controls were recruited before the COVID-19 pandemic (between 2019 and 2020) from the community through advertisement and from the endoscopy centre at the Prince of Wales Hospital in subjects who had a normal colonoscopy (stools collected before bowel preparation). Aged- and gender-matched controls with similar co-morbidities and standard dietary pattern were selected for comparison of gut microbiota composition between subjects with and without COVID-19 infection. Demographics and co-morbidities of controls were listed in Table 8. The exclusion criteria for non-COVID-19 controls were (1) the use of laxatives or anti-diarrheal drugs in the last 3 months; (2) recent dietary changes (e.g., becoming vegetarian/vegan); (3) known complex infections or sepsis; (4) known history of severe organ failure (including decompensated cirrhosis, malignant disease, kidney failure, epilepsy, active serious infection, acquired immunodeficiency syndrome); (5) bowel surgery in the last 6 months (excluding colonoscopy/procedure related to perianal disease); (6) presence of an ileostomy/stoma; and (7) current pregnancy. Given that diet is known to impact the gut microbiome, dietary records were documented for all COVID-19 patients during the time of hospitalisation. Standardized meals daily were provided by the department of hospital catering service of each hospital and the dietary component and pattern were consistent with the habitual diet commonly consumed by Hong Kong Chinese. After discharge, COVID-19 patients were advised to continue a diverse and standard Chinese diet that were consistent with habitual daily diets consumed by Hong Kong Chinese. The study was conducted in accordance with the declaration of Helsinki. All patients provided written informed consent. The study was approved by the Joint Chinese University of Hong Kong—New Territories East Cluster Clinical Research Ethics Committee (CREC Reference no.: 2020.076).
Detailed methods are described in Zuo et al16. Briefly, DNA was extracted from 0.1 g of homogenised faecal samples using the Maxwell RSC PureFood GMO and Authentication Kit and a Maxwell RSC instrument nucleic acid extraction platform (Promea, Wis., USA) according to manufacturer's instructions. Sequencing libraries Were prepared from extracted DNA using the Nextera DNA Flex Library Prep Kit (Illumina, Calif., USA) and sequenced on an Illurnina NovaSeq 6000 System (2×150 bp) at the Centre for Gut Microbiota Research, Chinese University of Hong Kong. Raw sequence data generated for this study are available in the Sequence Read Archive under BioProject accession: PRJNA714459.
Raw sequence data were quality filtered using Trimmomatic V.39 to remove adaptor and low-quality sequences and decontaminated against human genome (Reference: hg38) by Kneaddata (V.0.7.2 https://bitbucket.org/biobakery/kneaddata/wiki/Home). Following this, microbiota composition profiles were inferred from quality-filtered forward reads using MetaPhlAn3 version 3.0.5. GNU parallel17 was used for parallel analysis jobs to accelerate data processing.
Continuous variables were expressed in median (interquartile range) whereas categorical variables were presented as number (percentage). Qualitative and quantitative differences between subgroups were analysed using chi-squared or Fishers exact tests for categorical parameters and Mann-Whitney test for continuous parameters, as appropriate, Odds ratio and adjusted odds ratio (aOR) with 95% confidence interval (CI) were estimated using logistic regression to examine clinical parameters associated with development of PACS. The site by species counts and relative abundance tables were input into R V.3.5.1 for statistical analysis. Principal Coordinates Analysis (PCoA) was used to visualise the clustering of samples based on their species level compositional profiles. Associations between gut community composition and patients' parameters were assessed using permutational multivariate analysis of variance (PERMANOVA). Associations of specific microbial species with patient parameters were identified using the linear discriminant analysis effect size (LEfSe) and the multivariate analysis by linear models (MaAsLin) statistical frameworks implemented in the Huttenhower Lab Galaxy instance (website: huttenhower.sph.harvard.edu/galaxy/). PCoA, PERMANOVA and Procrustes analysis are implemented in the vegan R package V.2.5-7.
A random forest was executed to create a predictive model for PACS using gut microbiome composition at admission. The machine learning model analysed the relative abundance of microbiome taxonomic species-level inferred by MetaPhlAn3 version 3.0.5. Machine learning experiments used random forest, as this algorithm has been shown to outperform other learning tools for microbiome data18. For the implementation of the model, the scikit-learn implementation19 was used. An ensemble of 2,000 estimator trees and Shannon entropy were used to evaluate the quality of a split at each node of a tree. The two hyperparameters for the minimum number of samples per leaf and for the number of features per tree were set, as indicated elsewhere, to 5 and 30%, respectively. Hyperparameters for all methods were optimized by maximization of accuracy via 1.0-fold cross-validation.
All patents, patent applications, and other publications, including GenBank Accession Numbers and equivalents, cited in this application are incorporated by reference in the entirety for all purposes.
Agathobaculum
—
butyriciproducens
Bifidobacterium
—
pseudocatenulatum
Roseburia
—
hominis
Anaerostipes
—
hadrus
Dialister_sp_CAG_357
Eubacterium
—
rectale
Collinsella
—
aerofaciens
Gemmiger
—
formicilis
Firmicutes
—
bacterium_CAG_83
Coprococcus
—
comes
Faecalibacterium
—
prausnitzii
Eubacterium_sp_CAG_274
Ruthenibacterium
—
lactatiformans
Clostridium
—
leptum
Alistipes
—
putredinis
Eubacterium
—
ventriosum
Oscillibacter_sp_57_20
Fusicatenibacter
—
saccharivorans
Eubacterium
—
hallii
Dorea
—
formicigenerans
Asaccharobacter
—
celatus
Adlercreutzia
—
equolifaciens
Ruminococcus
—
torques
Roseburia
—
inulinivorans
Ruminococcus
—
lactaris
Ruminococcus
—
bicirculans
Barnesiella
—
intestinihominis
Eubacterium
—
ramulus
Lactococcus
—
petauri
Christensenella minuta
Eubacterium ramulus
Bifidobacterium adolescentis
Collinsella aerofaciens
Bifidobacterium pseudocatenulatum
Eubacterium rectale
Faecalibacterium prausnitzii
Fusobacterium ulcerans
Dialister sp. CAG: 357
Fusicatenibacter saccharivorans
Streptococcus anginosus group
Lactococcus petauri
Staphylococcus aureus
Rothia aeria
Collinsella stercoris
Blautia obeum
Roseburia faecis
Lawsonibacter asaccharolyticus
Ruminococcus torques
Coprococcus comes
Dorea longicatena
Lactobacillus fermentum
Actinomyces odontolyticus
Adlercreutzia equolifaciens
Agathobaculum butyriciproducens
Ruminococcus bicirculans
Megasphaera elsdenii
Romboutsia ilealis
Abiotrophia defectiva
Dorea formicigenerans
Clostridium citroniae
Firmicutes bacterium CAG: 83
Ruminococcus lactaris
Eubacterium sp. CAG: 38
Abiotrophia sp. HMSC24B09
Gemmiger formicilis
Eubacterium sp. CAG: 251
Clostridium leptum
Clostridium symbiosum
Roseburia intestinalis
Actinomyces sp. oral taxon 180
Phascolarctobacterium succinatutens
Barnesiella intestinihominis
Asaccharobacter celatus
Roseburia hominis
Enterorhabdus caecimuris
Enterococcus avium
Roseburia inulinivorans
Parabacteroides sp. CAG: 409
Parabacteroides goldsteinii
Coprococcus catus
Oscillibacter sp. 57_20
Actinomyces sp. oral taxon 181
Blautia wexlerae
Bifidobacterium longum
Eubacterium hallii
Sellimonas intestinalis
Ruminococcus bromii
Blautia sp. CAG: 257
Clostridium spiroforme
The above compositions typically comprise no detectable amount of any bacterial species in Table 4.
Lachnospira pectinoschiza
Scardovia wiggsiae
Oscillibacter sp. CAG: 241
Streptococcus vestibularis
Lactococcus garvieae
Erysipelatoclostridium ramosum
Stomatobaculum longum
Staphylococcus epidermidis
Holdemania filiformis
Clostridium innocuum
Parascardovia denticolens
Clostridium bolteae CAG 59
Fusobacterium varium
Clostridium bolteae
Scardovia inopinata
Bacteroides caccae
Rothia mucilaginosa
Streptococcus mutans
Ruminococcus gnavus
Actinomyces graevenitzii
Parabacteroides distasonis
Bacteroides vulgatus
Anaeromassilibacillus sp. An250
Massiliomicrobiota timonensis
Lactobacillus acidophilus
Peptostreptococcus anaerobius
Butyrivibrio sp. CAG: 318
Lactobacillus plantarum
Megamonas rupellensis
Lactobacillus johnsonii
Bifidobacterium animalis
Morganella morganii
Lactobacillus delbrueckii
Blautia producta
Turicimonas muris
Clostridiales bacterium 1_7_47FAA
Bacteroides thetaiotaomicron
Bacteroides sp. CAG: 144
Streptococcus lutetiensis
Butyricicoccus pullicaecorum
Blautia coccoides
Anaerotignum lactatifermentans
Megasphaera micronuciformis
Bacteroides xylanisolvens
Clostridium clostridioforme
Pediococcus acidilactici
Enterococcus casseliflavus
Intestinibacter bartlettii
Flavonifractor plautii
Actinomyces johnsonii
Atopobium parvulum
Bifidobacterium
—
adolescentis
Ruminococcus
—
gnavus
Bifidobacterium
—
pseudocatenulatum
Collinsella
—
aerofaciens
Bifidobacterium
—
longum
Bacteroides
—
vulgatus
Anaerostipes
—
hadrus
Fusicatenibacter
—
saccharivorans
Escherichia
—
coli
Bacteroides
—
uniformis
Blautia
—
wexlerae
Eubacterium
—
rectale
Faecalibacterium
—
prausnitzii
Dorea
—
longicatena
Klebsiella
—
pneumoniae
Roseburia
—
inulinivorans
Ruminococcus
—
bromii
Streptococcus
—
salivarius
Prevotella
—
copri
Eubacterium
—
hallii
Lachnospira
—
pectinoschiza
Scardovia
—
wiggsiae
Oscillibacter_sp_CAG_241
Streptococcus
—
vestibularis
Lactococcus
—
garvieae
Erysipelatoclostridium
—
ramosum
Stomatobaculum
—
longum
Staphylococcus
—
epidermidis
Holdemania
—
filiformis
Clostridium
—
innocuum
Parascardovia
—
denticolens
Clostridium
—
bolteae_CAG_59
Ruthenibacterium
—
lactatiformans
Fusobacterium
—
varium
Allisonella
—
histaminiformans
Eubacterium_sp_CAG_274
Clostridium
—
bolteae
Scardovia
—
inopinata
Oxalobacter
—
formigenes
Bacteroides
—
caccae
Rothia
—
mucilaginosa
Firmicutes
—
bacterium_CAG_110
Streptococcus
—
mutans
Ruminococcus
—
gnavus
Actinomyces
—
graevenitzii
Parabacteroides
—
distasonis
Bacteroides
—
vulgatus
Anaeromassilibacillus_sp_An250
Christensenella
—
minuta
Eubacterium
—
ramulus
Hungatella
—
hathewayi
Bifidobacterium
—
adolescentis
Collinsella
—
aerofaciens
Bifidobacterium
—
pseudocatenulatum
Eubacterium
—
rectale
Faecalibacterium
—
prausnitzii
Fusobacterium
—
ulcerans
Dialister_sp_CAG_357
Fusicatenibacter
—
saccharivorans
Streptococcus
—
anginosus_group
Lactococcus
—
petauri
Staphylococcus
—
aureus
Lactobacillus
—
mucosae
Rothia
—
aeria
Collinsella
—
stercoris
Blautia
—
obeum
Roseburia
—
faecis
Lawsonibacter
—
asaccharolyticus
Ruminococcus
—
torques
Coprococcus
—
comes
Dorea
—
longicatena
Lactobacillus
—
fermentum
Actinomyces
—
odontolyticus
Adlercreutzia
—
equolifaciens
Agathobaculum
—
butyriciproducens
Ruminococcus
—
bicirculans
Megasphaera
—
elsdenii
Romboutsia
—
ilealis
Abiotrophia
—
defectiva
Clostridium
—
aldenense
Dorea
—
formicigenerans
Clostridium
—
citroniae
Firmicutes
—
bacterium_CAG_83
Ruminococcus
—
lactaris
Eubacterium_sp_CAG_38
Clostridium
—
lavalense
Abiotrophia_sp_HMSC24B09
Gemmiger
—
formicilis
Eubacterium_sp_CAG_251
Clostridium
—
leptum
Clostridium
—
symbiosum
Roseburia
—
intestinalis
Actinomyces_sp_oral_taxon_180
Phascolarctobacterium
—
succinatutens
Eubacterium
—
ventriosum
Barnesiella
—
intestinihominis
Asaccharobacter
—
celatus
Roseburia
—
hominis
Enterorhabdus
—
caecimuris
Oxalobacter
—
formigenes
Collinsella
—
stercoris
Eubacterium_sp_CAG_274
Enterococcus
—
avium
Roseburia
—
inulinivorans
Parabacteroides_sp_CAG_409
Dorea
—
longicatena
Actinomyces_sp_S6_Spd3
Parabacteroides
—
goldsteinii
Ruminococcus
—
lactaris
Coprococcus
—
catus
Eubacterium_sp_CAG_251
Allisonella
—
histaminiformans
Enterorhabdus
—
caecimuris
Oscillibacter_sp_57_20
Ruthenibacterium
—
lactatiformans
Roseburia
—
faecis
Eubacterium
—
ramulus
Eubacterium
—
rectale
Clostridium
—
leptum
Actinomyces_sp_oral_taxon_181
Dorea
—
formicigenerans
Barnesiella
—
intestinihominis
Collinsella
—
aerofaciens
Ruminococcus
—
bicirculans
Firmicutes
—
bacterium_CAG_110
Firmicutes
—
bacterium_CAG_83
Asaccharobacter
—
celatus
Agathobaculum
—
butyriciproducens
Roseburia
—
hominis
Adlercreutzia
—
equolifaciens
Coprococcus
—
comes
Blautia
—
obeum
Gemmiger
—
formicilis
Ruminococcus
—
torques
Faecalibacterium
—
prausnitzii
Massiliomicrobiota
—
timonensis
Lactobacillus
—
acidophilus
Peptostreptococcus
—
anaerobius
Butyrivibrio_sp_CAG_318
Lactobacillus
—
plantarum
Megamonas
—
rupellensis
Lactobacillus
—
johnsonii
Bifidobacterium
—
animalis
Morganella
—
morganii
Lactobacillus
—
delbrueckii
Blautia
—
producta
Clostridium
—
lavalense
Turicimonas
—
muris
Clostridiales
—
bacterium_1_7_47FAA
Bacteroides
—
thetaiotaomicron
Bacteroides_sp_CAG_144
Streptococcus
—
lutetiensis
Butyricicoccus
—
pullicaecorum
Blautia
—
coccoides
Clostridium
—
aldenense
Anaerotignum
—
lactatifermentans
Megasphaera
—
micronuciformis
Bacteroides
—
xylanisolvens
Clostridium
—
clostridioforme
Pediococcus
—
acidilactici
Enterococcus
—
casseliflavus
Parabacteroides
—
distasonis
Hungatella
—
hathewayi
Bacteroides
—
vulgatus
Eubacterium
—
ventriosum
Intestinibacter
—
bartlettii
Clostridium
—
innocuum
Flavonifractor
—
plautii
Fusobacterium
—
varium
Lactobacillus
—
mucosae
Ruminococcus
—
gnavus
Streptococcus
—
mutans
Clostridium
—
bolteae
Erysipelatoclostridium
—
ramosum
Asaccharobacter
—
celatus
Propionibacterium
—
acidifaciens
Klebsiella
—
pneumoniae
Actinomyces
—
johnsonii
Enterorhabdus
—
caecimuris
Actinobaculum_sp_oral_taxon_183
Gemella
—
morbillorum
Blautia
—
obeum
Klebsiella
—
quasipneumoniae
Streptococcus
—
mitis
Eubacterium_sp_CAG_38
Olsenella
—
uli
Ruminococcus
—
lactaris
Streptococcus
—
cristatus
Adlercreutzia
—
equolifaciens
Catabacter
—
hongkongensis
Actinomyces_sp_S6_Spd3
Erysipelatoclostridium
—
ramosum
Parabacteroides
—
distasonis
Fusobacterium
—
ulcerans
Bacteroides
—
vulgatus
Bacteroides
—
thetaiotaomicron
Flavonifractor
—
plautii
Bacteroides
—
fragilis
Clostridium
—
lavalense
Actinomyces
—
odontolyticus
Agathobaculum
—
butyriciproducens
Faecalibacterium
—
prausnitzii
Gemmiger
—
formicilis
Intestinibacter
—
bartlettii
Roseburia
—
hominis
Actinomyces
—
naeslundii
Erysipelatoclostridium
—
ramosum
Oscillibacter_sp_57_20
Actinobaculum_sp_oral_taxon_183
Bifidobacterium
—
pseudocatenulatum
Clostridium
—
leptum
Collinsella
—
aerofaciens
Lactobacillus
—
salivarius
Actinomyces
—
oris
Actinomyces_sp_oral_taxon_414
Anaerostipes
—
hadrus
Bifidobacterium
—
dentium
Dialister_sp_CAG_357
Eubacterium
—
rectale
Eubacterium
—
ventriosum
Olsenella
—
uli
Streptococcus
—
anginosus_group
Actinomyces_sp_oral_taxon_448
Eubacterium_sp_CAG_274
Rothia
—
mucilaginosa
Streptococcus
—
infantis
Streptococcus
—
vestibularis
Actinomyces
—
massiliensis
Corynebacterium
—
durum
Firmicutes
—
bacterium_CAG_83
Ruthenibacterium
—
lactatiformans
Streptococcus
—
gordonii
Coprococcus
—
comes
Faecalicoccus
—
pleomorphus
Alistipes
—
putredinis
Actinomyces_sp_oral_taxon_897
Propionibacterium
—
acidifaciens
Dorea
—
formicigenerans
Lactobacillus
—
mucosae
Anaerostipes
—
caccae
Eubacterium
—
hallii
Fusicatenibacter
—
saccharivorans
Megasphaera
—
micronuciformis
Streptococcus
—
parasanguinis
Adlercreutzia
—
equolifaciens
Clostridium
—
innocuum
Asaccharobacter
—
celatus
Ruminococcus
—
gnavus
Eisenbergiella
—
tayi
Eubacterium
—
limosum
Blautia_sp_CAG_257
Ruminococcus
—
torques
Blautia
—
producta
Streptococcus
—
mutans
Actinomyces_sp_ICM47
Clostridium
—
disporicum
Clostridium
—
bolteae
Bacteroides
—
massiliensis
Bacteroides
—
vulgatus
Barnesiella
—
intestinihominis
Eubacterium
—
ramulus
Lactococcus
—
petauri
Romboutsia
—
ilealis
Ruminococcus
—
bicirculans
Actinomyces
—
graevenitzii
Pseudopropionibacterium
—
propionicum
Streptococcus
—
cristatus
Veillonella
—
parvula
Hungatella
—
hathewayi
Roseburia
—
inulinivorans
Ruminococcus
—
lactaris
Bacteroides
—
thetaiotaomicron
Eubacterium
—
callanderi
Haemophilus
—
parainfluenzae
Eisenbergiella
—
massiliensis
Faecalitalea
—
cylindroides
Firmicutes
—
bacterium_CAG_94
Lactonifactor
—
longoviformis
Massiliomicrobiota
—
timonensis
Pseudoflavonifractor_sp_An184
Actinomyces
—
naeslundii
Intestinibacter
—
bartlettii
Actinomyces
—
oris
Actinomyces_sp_ICM47
Actinomyces
—
odontolyticus
Streptococcus
—
anginosus_group
Streptococcus
—
vestibularis
Streptococcus
—
gordonii
Rothia
—
mucilaginosa
Clostridium
—
disporicum
Actinobaculum_sp_oral_taxon_183
Erysipelatoclostridium
—
ramosum
Veillonella
—
parvula
Streptococcus
—
mutans
Streptococcus
—
infantis
Lactobacillus
—
mucosae
Bifidobacterium
—
dentium
Actinomyces
—
graevenitzii
Olsenella
—
uli
Faecalicoccus
—
pleomorphus
Blautia
—
producta
Propionibacterium
—
acidifaciens
Actinomyces_sp_oral_taxon_448
Pseudopropionibacterium
—
propionicum
Actinomyces
—
massiliensis
Actinomyces_sp_oral_taxon_897
Streptococcus
—
cristatus
Actinomyces_sp_oral_taxon_414
Corynebacterium
—
durum
Ruminococcus
—
gnavus
Actinomyces
—
naeslundii
Clostridium
—
innocuum
Blautia_sp_CAG_257
Intestinibacter
—
bartlettii
Eubacterium
—
limosum
Actinomyces
—
odontolyticus
Actinobaculum_sp_oral_taxon_183
Lactobacillus
—
salivarius
Eisenbergiella
—
tayi
Erysipelatoclostridium
—
ramosum
Erysipelatoclostridium
—
ramosum
Ruminococcus
—
gnavus
Clostridium
—
innocuum
Actinomyces_sp_ICM47
Intestinibacter
—
bartlettii
Actinomyces
—
naeslundii
Actinomyces
—
oris
Bacteroides
—
vulgatus
Actinomyces
—
odontolyticus
Clostridium
—
bolteae
Streptococcus
—
mutans
Streptococcus
—
anginosus_group
Rothia
—
mucilaginosa
Streptococcus
—
gordonii
Bacteroides
—
massiliensis
Streptococcus
—
vestibularis
Bifidobacterium
—
dentium
Actinobaculum_sp_oral_taxon_183
Streptococcus
—
infantis
Clostridium
—
disporicum
Blautia
—
producta
Romboutsia
—
ilealis
Actinomyces_sp_oral_taxon_448
Actinomyces_sp_oral_taxon_414
Olsenella
—
uli
Corynebacterium
—
durum
Actinomyces
—
massiliensis
Intestinibacter
—
bartlettii
Clostridium
—
innocuum
Streptococcus
—
anginosus_group
Eisenbergiella
—
massiliensis
Hungatella
—
hathewayi
Faecalitalea
—
cylindroides
Eubacterium
—
limosum
Bifidobacterium
—
dentium
Clostridium
—
bolteae
Pseudoflavonifractor_sp_An184
Faecalicoccus
—
pleomorphus
Actinomyces
—
odontolyticus
Actinomyces
—
oris
Eisenbergiella
—
tayi
Lactonifactor
—
longoviformis
Firmicutes
—
bacterium_CAG_94
Olsenella
—
uli
Massiliomicrobiota
—
timonensis
Actinomyces_sp_oral_taxon_414
Intestinibacter
—
bartlettii
Hungatella
—
hathewayi
Streptococcus
—
anginosus_group
Clostridium
—
innocuum
Eubacterium
—
limosum
Actinomyces
—
oris
Ruminococcus
—
gnavus
Clostridium
—
bolteae
Bifidobacterium
—
dentium
Erysipelatoclostridium
—
ramosum
Bacteroides
—
thetaiotaomicron
Actinomyces
—
odontolyticus
Streptococcus
—
mutans
Streptococcus
—
vestibularis
Eisenbergiella
—
tayi
Haemophilus
—
parainfluenzae
Eubacterium
—
callanderi
Streptococcus
—
infantis
Actinomyces
—
naeslundii
Blautia
—
producta
Olsenella
—
uli
Actinomyces_sp_oral_taxon_448
Rothia
—
mucilaginosa
Faecalicoccus
—
pleomorphus
Lactobacillus
—
mucosae
Actinomyces_sp_oral_taxon_414
Actinomyces
—
naeslundii
Clostridium
—
innocuum
Actinomyces
—
oris
Intestinibacter
—
bartlettii
Actinomyces
—
odontolyticus
Streptococcus
—
parasanguinis
Bifidobacterium
—
dentium
Erysipelatoclostridium
—
ramosum
Streptococcus
—
anginosus_group
Megasphaera
—
micronuciformis
Rothia
—
mucilaginosa
Streptococcus
—
vestibularis
Anaerostipes
—
caccae
Streptococcus
—
gordonii
Actinobaculum_sp_oral_taxon_183
Lactobacillus
—
mucosae
Propionibacterium
—
acidifaciens
Actinomyces_sp_oral_taxon_448
Olsenella
—
uli
Streptococcus
—
infantis
Actinomyces_sp_oral_taxon_414
Faecalicoccus
—
pleomorphus
Actinomyces_sp_oral_taxon_897
Lactobacillus
—
salivarius
Actinomyces
—
massiliensis
Corynebacterium
—
durum
Bifidobacterium
—
longum
Bifidobacterium
—
pseudocatenulatum
Blautia
—
wexlerae
Faecalibacterium
—
prausnitzii
Fusicatenibacter
—
saccharivorans
Anaerostipes
—
hadrus
Eubacterium
—
hallii
Roseburia
—
inulinivorans
Dorea
—
longicatena
Coprococcus
—
comes
Firmicutes
—
bacterium_CAG_83
Streptococcus
—
salivarius
Agathobaculum
—
butyriciproducens
Roseburia
—
faecis
Sellimonas
—
intestinalis
Eubacterium
—
ramulus
Clostridium
—
disporicum
Actinomyces
—
naeslundii
Holdemania
—
filiformis
Phascolarctobacterium
—
faecium
This application claims priority to U.S. Provisional Patent Application No. 63/272,608, filed Oct. 27, 2021, the contents of which are hereby incorporated by reference in the entirety for all purposes.
| Number | Date | Country | |
|---|---|---|---|
| 63272608 | Oct 2021 | US |
