Microorganisms for use and delivery to the respiratory system

Abstract

This invention provides a formulation containing microorganisms for delivery in the form of a plurality of particles, a kit for delivering the microbial formulation in the form of a plurality of particles, and an apparatus for dispensing the microbial formulation in the form of a plurality of particles. A method for dispensing microorganisms via spray or inhalation is provided. A method for introducing microorganisms into the respiratory system is provided. A method for treating conditions, including respiratory conditions and inflammatory conditions, with a formulation containing microorganisms is provided.

Description

[Technical Field] The state of matter is broadly related to the microbial community, and more specifically, to the restoration of ammonia-oxidizing microorganisms or non-pathogenic bacteria, such as bacteria that are beneficial to the microbial community. [Previous Technology] Bacteria and other microorganisms are ubiquitous in the environment. The discovery of pathogens and theories of disease origin have a profound impact on health and disease states. Microorganisms are a normal and potentially beneficial part of the environment for all living organisms. For example, in the gut, bacteria are not pathogenic under normal conditions and in fact improve health by making normal intestinal contents less responsive to disease-causing organisms. [Summary of the Invention] According to one embodiment, a method for preparing, for example, a live in vitro non-pathogenic bacterial preparation is provided. The method for preparing the non-pathogenic bacterial preparation may include providing a quantity of a mixture comprising non-pathogenic bacteria. The method for preparing the non-pathogenic bacterial preparation may include subjecting the mixture to conditions that form a plurality of particles in the mixture, thereby preparing the live in vitro non-pathogenic bacterial preparation. In some embodiments, the particle size range (PSR) is about 5, 50, 250, or 500 micrometers or less. In some embodiments, the method may include administering a non-pathogenic agent to an individual. According to one embodiment, a method for introducing non-pathogenic bacteria into an individual is provided. The method for introducing non-pathogenic bacteria may include forming a plurality of particles comprising a preparation containing non-pathogenic bacteria. In some embodiments, the particle size range (PSR) of the particles is about 5, 50, 250, or 500 micrometers or less. The method for introducing non-pathogenic bacteria may further include delivering the plurality of particles into the individual's respiratory system. According to one embodiment, a method for supplementing or modulating an individual's microbial community is provided. The individual's microbial community may be the individual's lung microbial community. The method for supplementing or modulating an individual's microbial community may include administering an agent containing non-pathogenic bacteria into the individual's respiratory system, thereby supplementing or modulating the individual's microbial community. In some embodiments, the agent may contain a plurality of particles with a particle size range (PSR) of approximately 5, 50, 250, or 500 micrometers or smaller. In some embodiments, the method may include administering non-pathogenic bacteria via spraying. According to one example, a method for introducing non-pathogenic bacteria into an individual is provided. The method for introducing non-pathogenic bacteria into an individual may include administering an agent containing non-pathogenic bacteria into the individual's respiratory system, for example, via spraying. In some embodiments, the method may further include providing a quantity of a mixture containing non-pathogenic bacteria. The method may include subjecting the mixture to conditions that form a plurality of particles. The PSR of the plurality of particles in the mixture may be about 5, 50, 250, or 500 micrometers or less. In some embodiments, the method may include providing the mixture of non-pathogenic bacteria and separating that quantity from the mixture. According to some embodiments, the mixture may further comprise a carrier. The carrier may comprise a liquid or a gas. The mixture may further comprise a therapeutic agent. In some embodiments, the mixture may further comprise a steroid. Non-pathogenic bacteria can be administered via inhalation or intratracheal delivery. In some embodiments, each of the plurality of particles may comprise a droplet. The particles may be delivered to an individual at a flow rate of about 0.5 mL / min or less. For example, the particles may be delivered to an individual at a flow rate of about 0.2 mL / min. The conditions for forming the plurality of particles may include mechanical force or electricity. In some embodiments, at least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the amount can be converted into particles within the PSR. At least 99, 99.9, 99.99, or 99.999% of the amount can be converted into particles within the PSR. In some embodiments, substantially all of the amount can be converted into particles within the PSR. According to some embodiments, the PSR of the particles can be between about 1 micrometer and about 5 micrometers. At least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the particles have a PSR between about 1 micrometer and about 5 micrometers. At least 99, 99.9, 99.99, or 99.999% of the particles have a PSR between about 1 micrometer and about 5 micrometers. In some embodiments, substantially all particles have a PSR between about 1 micrometer and about 5 micrometers. The PSR of the particles can be between about 1 micrometer and about 10 micrometers. At least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the particles have a PSR between about 1 micrometer and about 10 micrometers. At least 99, 99.9, 99.99, or 99.999% of the particles have a PSR between about 1 micrometer and about 10 micrometers. In some embodiments, substantially all particles have a PSR between about 1 micrometer and about 10 micrometers. In some embodiments, the PSR of the particles can be between about 1 micrometer and about 50 micrometers. At least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the particles have a PSR between about 1 micrometer and about 50 micrometers. At least 99, 99.9, 99.99, or 99.999% of the particles have a PSR between about 1 micrometer and about 50 micrometers. In some embodiments, substantially all particles have a PSR between about 1 micrometer and about 50 micrometers. According to some embodiments, the PSR of the particles can be between about 1 micrometer and about 100 micrometers. At least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the particles have a PSR between about 1 micrometer and about 100 micrometers. At least 99, 99.9, 99.99, or 99.999% of the particles have a PSR between about 1 micrometer and about 100 micrometers. In some embodiments, substantially all particles have a PSR between about 1 micrometer and about 100 micrometers. The PSR of the particles can be between about 5 micrometers and about 50 micrometers. At least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the particles have a PSR between about 5 micrometers and about 50 micrometers. At least 99, 99.9, 99.99, or 99.999% of the particles have a PSR between about 5 micrometers and about 50 micrometers. In some embodiments, substantially all particles have a PSR between about 5 micrometers and about 50 micrometers. In some embodiments, the PSR of the particles may be between about 10 micrometers and about 100 micrometers. At least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the particles may have a PSR between about 10 micrometers and about 100 micrometers. At least 99, 99.9, 99.99, or 99.999% of the particles may have a PSR between about 10 micrometers and about 100 micrometers. In some embodiments, substantially all particles may have a PSR between about 10 micrometers and about 100 micrometers. According to some embodiments, non-pathogenic bacteria can be purified. Non-pathogenic bacteria may be live bacteria. In some embodiments, at least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the particles may contain live non-pathogenic bacteria. At least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the non-pathogenic bacteria in each of the plurality of particles may be live bacteria. At least 99, 99.9, 99.99, or 99.999% of the non-pathogenic bacteria in each of the plurality of particles may be live bacteria. In some embodiments, substantially all of the non-pathogenic bacteria in each of the plurality of particles may be live bacteria. At least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the non-pathogenic bacteria in each of the plurality of particles may be non-pathogenic. At least 99, 99.9, 99.99, or 99.999% of the non-pathogenic bacteria in each of the plurality of particles may be non-pathogenic. In some embodiments, substantially all of the non-pathogenic bacteria in each of the plurality of particles may be non-pathogenic. In some embodiments, at least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the non-pathogenic bacteria in each of the plurality of particles may consist substantially of isolated species. At least 99, 99.9, 99.99, or 99.999% of the non-pathogenic bacteria in each of the plurality of particles may consist substantially of isolated species. In some embodiments, substantially all of the non-pathogenic bacteria in each of the plurality of particles may consist substantially of isolated species. The isolated species can be selected from the following non-pathogenic species groups: *Prevotella*, *Sphingomonas*, *Pseudomonas*, *Acinetobacter*, *Fusobacterium*, *Megasphaera*, *Veillonella*, *Staphylococcus*, *Streptococcus*, and *St.* The species may include *Corynebacterium*, *Propionibacterium*, *Rhodococcus*, *Microbacterium*, *Streptococcus*, *Bacillus*, *Lactobacillus*, *Lactococcus*, *Streptomyces*, *Faecalibacterium*, *Bacteroides*, or *Bifidobacter*. In some embodiments, the isolated species may be *Nitrosomonas eutropha* (N. eutropha). In some embodiments, the isolated species may be *Nitrosomonas eutropha* D23 with ATCC accession number PTA-121157. In some embodiments, at least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the non-pathogenic bacteria in each of the plurality of particles may consist substantially of a selected bacterial species community. At least 99, 99.9, 99.99, or 99.999% of the non-pathogenic bacteria in each of the plurality of particles may consist substantially of a selected bacterial species community. In some embodiments, substantially all of the non-pathogenic bacteria in each of the plurality of particles may consist substantially of a selected bacterial species community. In some embodiments, the selected bacterial community may include species such as *Prevotella*, *Sphingomonas*, *Pseudomonas*, *Acinetobacter*, *Clostridium*, *Macrococcus*, *Vanillococcus*, *Staphylococcus*, or *Streptococcus*, and combinations thereof. In some embodiments, the selected bacterial community may include species such as Staphylococcus, Corynebacterium, Propionibacterium, Rhodococcus, Microbacterium, or Streptococcus and combinations thereof. In some embodiments, the selected bacterial community may include species such as Bacillus, Lactobacillus, Lactococcus, Streptomyces, Faecalibacterium, Bacteroides, or Bifidobacterium and combinations thereof. According to certain embodiments, non-pathogenic bacteria may include species such as Prevotella, Sphingomonas, Pseudomonas, Acinetobacter, Clostridium, Megacoccus, Vanilloidococcus, Staphylococcus, Streptococcus, Corynebacterium, Propionibacterium, Rhodococcus, Microbacterium, Bacillus, Lactobacillus, Lactococcus, Streptococcus, Faeobacterium, Bacteroides, or Bifidobacterium and combinations thereof. In some embodiments, at least 10, 20, 50, 70, 80, 90, or 95% of the non-pathogenic bacteria in each of the plurality of particles may be selected from species comprising the group consisting of: *Prevotella*, *Sphingomonas*, *Pseudomonas*, *Acinetobacter*, *Clostridium*, *Macrococcus*, *Vanillococcus*, *Staphylococcus*, *Streptococcus*, *Corynebacterium*, *Propionibacterium*, *Rhodococcus*, *Microbacterium*, *Bacillus*, *Lactobacillus*, *Lactococcus*, *Streptomyces*, *Femtobacter*, *Bacteroides*, or *Bifidobacterium* and combinations thereof. In some embodiments, the non-pathogenic bacteria may comprise species such as *Nitrosomonas*, *Nitrosococcus*, *Nitrosospira*, *Nitrosocystis*, *Nitrosolobus*, *Nitrosovibrio*, and combinations thereof. At least 10, 20, 50, 70, 80, 90, or 95% of the non-pathogenic bacteria in each of the plurality of particles may be selected from species comprising the group consisting of *Nitrosomonas*, *Nitrosococcus*, *Nitrosospira*, *Nitrosocystis*, *Nitrosolobus*, or *Nitrosovibrio*, and combinations thereof. Non-pathogenic bacteria may include ammonia-oxidizing bacteria (AOB). Non-pathogenic bacteria may include eutrophic nitrosomonas (N. eutropha). Non-pathogenic bacteria may include eutrophic nitrosomonas D23 with ATCC registration number PTA-121157. In some embodiments, AOB may be able to convert ammonia or ammonium to nitrite at a rate of at least about 1 pmol / min / mg protein, for example at least about 0.1 nanomol / min / mg protein. In some embodiments, non-pathogenic bacteria may be substantially free of AOB. According to some embodiments, non-pathogenic bacteria may be derived from biological samples. Non-pathogenic bacteria may comprise bacteria present in or found in mammalian microbiota, such as the human microbiota. In some embodiments, non-pathogenic bacteria may comprise a first bacterium present in or found in a mammalian microbiota, such as the human microbiota, and a second bacterium of a species other than the first bacterium present in or found in a mammalian microbiota, such as the human microbiota. Non-pathogenic bacteria may comprise a community of bacterial species present in or found in a mammalian microbiota, such as the human microbiota. Non-pathogenic bacteria may include bacteria or bacterial species communities present in or found in the gastrointestinal microbiota of mammals, such as the oral, intestinal, colonic, or fecal microbiota. In some embodiments, non-pathogenic bacteria may include bacteria or bacterial species communities present in or found in the respiratory microbiota of mammals, such as the nasal cavity, trachea, or lung microbiota. Non-pathogenic bacteria may include bacteria or bacterial species communities present in or found in the skin microbiota of mammals. Non-pathogenic bacteria may include bacteria or bacterial species communities present in or found in the microbiota of healthy or non-disease-prone mammals, such as humans. In some embodiments, the non-pathogenic bacteria may be derived from a donor. The method may include selecting a donor. The method may include assessing the acceptability of the donor, such as assessing whether the donor meets predetermined criteria or references. In some embodiments, the method may include assessing whether the donor is healthy, such as physiological, normal, or disease-free. The method may include assessing the donor regarding a healthy microbiome, such as a physiological, normal, or disease-free microbiome. The method may include assessing the donor regarding the presence or absence of a condition or symptom, such as a respiratory condition or symptom. According to some embodiments, donors may be selected. Donors may be evaluated regarding acceptability, such as conformity to predetermined criteria or references. Donors may be evaluated regarding their health, such as whether they are physiological, normal, or disease-free. In some embodiments, donors may be evaluated regarding their healthy microbiome, such as a physiologically normal or disease-free microbiome. Donors may be evaluated regarding the presence or absence of a condition or symptom, such as a respiratory condition or symptom. The donor may be substantially free of disease or symptom, such as respiratory disease or symptom. In some embodiments, the donor may be substantially free of disease or symptom, such as symptoms of respiratory disease or symptom. In some embodiments, in response to donor evaluation, the donor may be accepted or rejected. In some embodiments, the biological sample may be obtained from the donor. The biological sample may be obtained from the donor in response to the donor evaluation. According to some embodiments, the method may include obtaining a biological sample from a donor. The method may include selecting a biological sample. In some embodiments, the method may include assessing the microbial community of the biological sample regarding acceptability, such as compliance with predetermined criteria or references. The method may include assessing the microbial community of the biological sample regarding a healthy microbial community, such as a physiologically normal or non-disease-prone microbial community. The method may include assessing the microbial community of the biological sample regarding the presence of pathogenic bacteria. In some embodiments, the method may include assessing the microbial community of the biological sample regarding a critical limit concentration of pathogenic bacteria. The microbial community of biological samples should be substantially free of pathogenic bacteria. The microbial community of biological samples should be substantially free of *M. catarrhalis*, *H. influenzae*, *S. pneumoniae*, or *S. aureus*, or combinations thereof. In some embodiments, the method may include assessing the microbial community of a biological sample based on the presence of Moraxella catarrhalis, Haemophilus influenzae, Streptococcus pneumoniae, or Staphylococcus aureus and combinations thereof. The method may also include assessing the microbial community of a biological sample based on critical limit concentrations of Moraxella catarrhalis, Haemophilus influenzae, Streptococcus pneumoniae, or Staphylococcus aureus and combinations thereof. In response to a biological sample assessment, the biological sample may be accepted or rejected. In some embodiments, non-pathogenic bacteria may be derived from the biological sample in response to the biological sample assessment. Depending on certain morphologies, an individual may possess a disrupted microbial community. An individual's microbial community, such as the gastrointestinal, respiratory, or skin microbiome, may contain pathogenic bacteria. An individual's microbial community, such as the gastrointestinal, respiratory, or skin microbiome, may contain Moraxella catarrhalis, Haemophilus influenzae, Streptococcus pneumoniae, Staphylococcus aureus, Vibrio cholerae, Escherichia coli, or species and combinations thereof from the genera Shigella, Campylobacter, or Salmonella. In some embodiments, the method may include administering non-pathogenic bacteria, such as non-pathogenic bacteria derived from a donor, to an individual. In some embodiments, the non-pathogenic bacteria derived from the donor may compete with pathogenic bacteria in the individual's microbiome. The target percentage of bacteria can be transferred to an individual's respiratory system. In some embodiments, the method further includes treating an individual's respiratory condition. Treating the respiratory condition may include reducing inflammation. In some embodiments, the respiratory condition may be an inflammatory condition. The respiratory condition may be an airway disease. The respiratory condition may be a heart, vascular, or lung condition. In some embodiments, the respiratory condition may be asthma, allergies, carbon monoxide poisoning, smoke inhalation, emphysema, asbestos poisoning, bronchitis, pulmonary fibrosis, cystic fibrosis, embolism, chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome, pulmonary hypertension, Celiac's disease, or pneumonia. According to some embodiments, the formulation may be administered to a first tissue. The formulation may be administered via inhalation. The first tissue may be a depositing tissue. In some embodiments, the first tissue may be a target tissue. In other embodiments, the first tissue may be a tissue other than a target tissue. The formulation may be applied to the first tissue and the formulation or formulation product may be delivered to a second tissue. The formulation or formulation product may be delivered by diffusion. In some embodiments, the second tissue is a target tissue. In some embodiments, the deposited tissue, target tissue, or both may be associated with an individual's nose, nasopharynx, larynx, or trachea. Target tissue may be associated with the trachea (air duct), larynx, pharynx, small trachea, segmental bronchus, subsegmental bronchus, lung apex, pleura, pleural cavity, alveolar ducts, alveoli, main bronchus, lobar bronchus, hilum, upper lobe of the lung (including apical, posterior, and anterior segments), middle lobe of the lung (including medial basal and lateral segments), or lower lobe of the lung (including upper, posterior, or anterior basal segments). The target tissue may be associated with the desired local effect. In some embodiments, the desired local effect may involve treating infection, inflammation, or congestion of the trachea, lungs, nasal passages, or other respiratory tissues. In some embodiments, the desired local effect may be treating asthma, allergies, carbon monoxide poisoning, smoke inhalation, emphysema, asbestos poisoning, bronchitis, pulmonary fibrosis, cystic fibrosis, embolism, chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome, pulmonary hypertension, or pneumonia. The target tissue may be associated with the desired systemic effect. The desired systemic effect may include treatment of one or more of the following: headache, cardiovascular disease, inflammation, immune response, autoimmune disease, liver disease, infection, neurological disease, psychiatric disease, nitric oxide disorder, urea cycle disorder, congestion, vasodilatory disorder, skin disease, ophthalmic disease, wound healing, insect bite reaction, connective tissue disorder, and certain viral, bacterial, or fungal infections. In some embodiments, administration of an effective amount of the formulation may promote endothelial function. Administering an effective dose of the preparation can promote the colonization of non-pathogenic bacteria in tissues. These tissues can be respiratory system tissues. In some embodiments, administration of an effective amount of the formulation can modulate an individual's maximum VO2. Administration of an effective amount of the formulation can modulate an individual's cardiac metabolic rate. In some embodiments, administration of an effective amount of the formulation can modulate an individual's microbiome. The microbiome may be associated with an individual's respiratory system. The microbiome may be associated with an individual's gastrointestinal system. The microbiome may be associated with an individual's skin. In some embodiments, administration of an effective amount of the formulation may alter or change an individual's nitrite or NO levels. Administration of an effective amount may alter or change nitrite or NO levels at a target tissue site in an individual. Administration of an effective amount may alter or change nitrite or NO levels systemically. The administration of the formulation can be done via an auxiliary device. According to some embodiments, the formulation may be administered before the onset of a respiratory condition. In some embodiments, the formulation may be administered during the onset of a respiratory condition. In some embodiments, the formulation may be administered after the respiratory condition has subsided. The formulation may be administered in response to respiratory condition symptoms, triggers, or warning signals. For example, the formulation may be administered in response to cough or dyspnea. In some embodiments, the method may include determining whether an individual needs to be given a formulation. The method may also include determining whether an individual needs treatment for a respiratory condition. The formulation can be administered in solution, suspension, or liquid form. It can also be administered as droplets, sprays, aerosols, or mists. In some embodiments, the formulation may be formulated as a respiratory solution (e.g., ultrafine droplets, aerosol, or aerosol), gas, or dry powder. The formulation may include microspheres or microcapsules. In some embodiments, the formulation may be formulated to be compatible with an individual's respiratory system. The formulation may be substantially isotonic. The formulation may have a substantially physiological pH level. In some embodiments, the formulation may be formulated for immediate release. The formulation may be formulated for prolonged release. The formulation may be formulated for delivery of non-pathogenic bacteria to a target tissue. The formulation may be formulated for delivery of a product of non-pathogenic bacteria to a target tissue. The target tissue may be local or systemic. In some embodiments, the formulation may be formulated for local or systemic delivery of nitrite or NO to a target tissue. The formulation may be formulated for mucosal delivery and / or circulation. Mucosal delivery and / or circulation may be local or systemic. According to some embodiments, the method may additionally include administering a second amount of the formulation to an individual. According to some embodiments, the formulation may be administered as part of a combination therapy. The method may additionally include administering a second treatment in combination with the formulation. The formulation may be administered for a period of time prior to initiation of the second treatment. The formulation may be administered concurrently with the second treatment. The formulation may be administered for a period of time after cessation of the second treatment. In some embodiments, the second treatment may be administered via an alternating dosing pattern. For example, the second treatment may be administered orally or intranasally. An individual may receive a second treatment at a therapeutic level. In some embodiments, the second treatment includes a surgical procedure. The preparation may be administered before or after the surgical or diagnostic procedure. In some embodiments, the formulation may be administered in combination with an anti-inflammatory agent. The formulation may be administered in combination with a medical method for treating an associated disease or condition, or symptoms of an associated disease or condition. The formulation may be administered in combination with a medical method approved for treating an associated disease or condition, or symptoms of an associated disease or condition. The formulation may be administered in combination with a medical method commonly used for treating an associated disease or condition, or symptoms of an associated disease or condition. The formulation may be administered in combination with asthma medications, cold and flu medications, corticosteroids, or antihistamines. It may also be administered in combination with therapeutic treatments for asthma, allergies, carbon monoxide poisoning, asbestos poisoning, bronchitis, pulmonary fibrosis, cystic fibrosis, embolism, COPD, adult respiratory distress syndrome, pulmonary hypertension, or Cesarean disease. The formulation may be administered in combination with products of non-pathogenic bacteria. The product may also be administered in combination with compounds that promote the growth or metabolism of non-pathogenic bacteria. According to some embodiments, the effective amount may be a therapeutically effective dose for non-pathogenic bacteria. The therapeutically effective dose for non-pathogenic bacteria may be approximately or greater than about 1×10³, 10⁴, 10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰, 10¹¹, 10¹², 10¹³, or 10¹⁴ CFU. In some embodiments, the formulation may be administered as an analgesic. In some embodiments, the formulation may be administered as a prophylactic agent. In some embodiments, the formulation may be self-administered. The formulation can be administered approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 times daily. Each administration can be given over a period of approximately less than 1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45 minutes or more. The formulation can be administered for approximately 1-3, 3-5, 5-7, 7-9, 5-10, 10-14, 12-18, 12-21, 21-28, 28-35, 35-42, 42-49, 49-56, 46-63, 63-70, 70-77, 77-84, or 84-91 days. The formulation can be administered daily, every 2, 3, 4, 5, 6 days, weekly, or bi-weekly. The formulation can be administered as needed. The preparation can be administered to the individual 30, 60, 90, 120, 150, or 180 minutes after waking from sleep. The preparation can be administered to the individual 30, 60, 90, 120, 150, or 180 minutes before going to sleep. The preparation can be administered to the individual 30, 60, 90, 120, 150, or 180 minutes after eating. The preparation can be administered to the individual 30, 60, 90, 120, 150, or 180 minutes before cleaning or showering. The preparation can be administered to the individual 30, 60, 90, 120, 150, or 180 minutes after cleaning or showering. In some embodiments, the individual may be female. In some embodiments, the individual may be male. The individual may be characterized as one of the following races / ethnicities: Asian, Black or African American, Hispanic or Latino, White, or multiracial. The individual may be less than 1 year old, or between 1-5, 5-10, 10-20, 20-30, 30-40, 40-50, 50-60 years old, or over 60 years old. According to some embodiments, the formulation may contain a buffer solution containing non-pathogenic bacteria. The formulation may contain an aqueous buffer solution containing non-pathogenic bacteria. The buffer solution may contain disodium hydrogen phosphate and magnesium chloride, for example, water containing 50 mM Na₂HPO₄ and 2 mM MgCl₂. The buffer solution may consist essentially of disodium hydrogen phosphate and magnesium chloride, for example, water containing 50 mM Na₂HPO₄ and 2 mM MgCl₂. The buffer solution may consist of disodium hydrogen phosphate and magnesium chloride, for example, water containing 50 mM Na₂HPO₄ and 2 mM MgCl₂. In some embodiments, the formulation may further comprise a compound that promotes the growth or metabolism of non-pathogenic bacteria. The formulation may be administered simultaneously with the compound that promotes the growth or metabolism of non-pathogenic bacteria. In some embodiments, the formulation may further comprise a product of non-pathogenic bacteria. The formulation may be administered simultaneously with the product of non-pathogenic bacteria. In some embodiments, the formulation may further comprise a compound that promotes NO production and / or urease activity. The formulation may be administered simultaneously with a compound that promotes NO production and / or urease activity. In some embodiments, the formulation may further comprise at least one of ammonia, an ammonium salt, and urea. The formulation may be administered simultaneously with at least one of ammonia, an ammonium salt, and urea. The formulation may further include a controlled-release material. For example, the formulation may further include a slow-release material. The formulation may further include excipients. For example, the formulation may further include pharmaceutically acceptable excipients. In some embodiments, the excipients may include absorption or permeation enhancers, preservatives, antioxidants, buffers, chelating agents, ion exchangers, solubilizers, suspending agents, thickeners, surfactants, wetting agents, tension modifiers, enzyme inhibitors, or mediators for appropriate drug delivery, such as propellants. The formulation may further include mucosal adhesives. The formulation may include disintegrants, chelating agents, coating agents, controlled-release products, or fillers. According to some embodiments, the formulation may be substantially free of other living organisms. The formulation may be substantially free of non-living substances. In some embodiments, the formulation may contain non-pathogenic bacteria between about 1×10³ CFU / mL and about 1×10¹⁴ CFU / mL. The formulation may contain non-pathogenic bacteria between about 1×10⁹ CFU / mL and about 10×10⁹ CFU / mL. In some embodiments, the biocommunity-friendly product may be used in combination with an administration formulation containing non-pathogenic bacteria. According to another embodiment, a mixture containing non-pathogenic bacteria is provided. The mixture may be formed into a plurality of particles. For example, the mixture may be formed into a plurality of particles with a particle size range (PSR) of about 5, 50, 250, or 500 micrometers or smaller. The mixture may further include a carrier. The carrier may be a liquid or a gas. In some embodiments, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the particles in the mixture may be within the PSR. In some embodiments, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the non-pathogenic bacteria in each of the plurality of particles in the mixture may be live bacteria. In some embodiments, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the bacteria in each of the plurality of particles in the mixture may be non-pathogenic bacteria. In some embodiments, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, or 99% of the non-pathogenic bacteria in each of the plurality of particles in the mixture may be selected from species comprising the group consisting of: *Prevotella*, *Sphingomonas*, *Pseudomonas*, *Acinetobacter*, *Clostridium*, *Macrococcus*, *Vanillococcus*, *Staphylococcus*, *Streptococcus*, *Corynebacterium*, *Propionibacterium*, *Rhodococcus*, *Microbacterium*, *Bacillus*, *Lactobacillus*, *Lactococcus*, *Streptomyces*, *Femtobacter*, *Bacteroides*, or *Bifidobacterium* and combinations thereof. In some embodiments, the non-pathogenic bacteria in the mixture include AOB. In some embodiments, the non-pathogenic bacteria in the mixture are substantially free of AOB. According to another approach, a preparation containing non-pathogenic bacteria is provided to an individual via spraying. According to another approach, a preparation containing non-pathogenic bacteria is provided to an individual via inhalation or intratracheal delivery. According to another embodiment, preparations containing non-pathogenic bacteria are provided for the treatment of respiratory conditions. Alternatively, preparations containing non-pathogenic bacteria are provided for modulating an individual's microbial community. The microbial community may be the individual's lung or nasal microbial community. In some embodiments, the formulation is prepared as a respiratory solution, gas, or dry particles. The formulation may be prepared as ultrafine droplets, aerosols, or aerosols. The formulation may be prepared for spraying. The formulation may be prepared for inhalation. The non-pathogenic bacteria in the formulation may include selected bacterial communities. The non-pathogenic bacteria in the formulation may be selected from species comprising the following groups: Prevotella, Sphingomonas, Pseudomonas, Acinetobacter, Clostridium, Megacoccus, Vanilloidococcus, Staphylococcus, Streptococcus, Corynebacterium, Propionibacterium, Rhodococcus, Microbacterium, Bacillus, Lactobacillus, Lactococcus, Streptococcus, Faeobacterium, Bacteroides, or Bifidobacterium and combinations thereof. The formulation can be packaged for single use. The formulation can be packaged for multiple uses. The formulation may further contain therapeutic agents. For example, the formulation may further contain steroids. According to another embodiment, a device is provided that is structurally designed to administer a preparation containing non-pathogenic bacteria. The device may be structurally designed to administer the preparation to deposited or target tissue. The deposited or target tissue may be within an individual's respiratory system. In some embodiments, the device is a nebulizer. In some embodiments, the device is an inhaler. According to another embodiment, an apparatus comprising a sprayer and a reservoir is provided. The sprayer can form droplets with a particle size range (PSR) of approximately 5, 50, 250, or 500 micrometers or smaller. The reservoir can hold a mixture containing non-pathogenic bacteria. The mixture may further contain a carrier. In some embodiments, the device may be mechanically assisted. In some embodiments, the device may be electrically assisted. The device can provide a quantitative dose of non-pathogenic bacteria. According to another embodiment, a kit comprising an apparatus and a formulation is provided. The apparatus may be structurally designed to administer a formulation containing non-pathogenic bacteria. The formulation may contain non-pathogenic bacteria. In some embodiments, the device included in the kit is a nebulizer. In some embodiments, the device included in the kit is an inhaler. In some embodiments, the devices in the kit are designed for multiple uses. The kit may contain more than one dose of a non-pathogenic preparation as described in any of the foregoing embodiments. The kit may further include instructions for use. The kit may include instructions for administering the formulation to an individual. For example, the kit may include instructions for administering the formulation to an individual via inhalation or intratracheal delivery. The kit may include instructions for providing a mixture containing non-pathogenic bacteria. The kit may include instructions for subjecting the mixture to conditions that form a plurality of particles. For example, the kit may include instructions for subjecting the mixture to conditions that form a plurality of particles with a particle size range (PSR) of approximately 5, 50, 250, or 500 micrometers or less. According to one embodiment, a method for introducing ammonia-oxidizing microorganisms (AOM) into an individual is provided. The method may include administering an AOM-containing preparation to the individual's respiratory system, for example, via inhalation or intratracheal delivery. In some embodiments, the method may include administering a preparation containing live AOM. According to one embodiment, a method for inoculating an individual's lungs with AOM is provided. The method for inoculating an individual's lungs may include administering an AOM-containing preparation into the individual's lungs, for example, wherein the AOM penetrates target lung tissue. In some embodiments, a target percentage of the administered AOM is transferred to the individual's respiratory system. For example, at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the administered AOM can be transferred to the individual's target deposited tissue. In some embodiments, the AOM penetrates the target lung tissue in the form of an aerosol. The droplet size of the aerosol formulation can be approximately 5, 50, 250, or 500 micrometers or smaller. According to one embodiment, a method for delivering nitric oxide (NO) to an individual is provided. The method of delivering NO may include administering an effective amount of a formulation comprising nebulized ammonia (AOM) to a target tissue of the individual. The target tissue may be the individual's respiratory system tissue. The nebulized AOM may be delivered via inhalation, a mask, or an intratracheal device, thereby delivering NO to the individual. In some embodiments, nitrite is also delivered to the individual. According to one example, a method for treating a respiratory ailment in an individual is provided. The method for treating a respiratory ailment may include administering an effective amount of a preparation containing AOM to the individual, thereby treating the respiratory ailment. In some embodiments, treating a respiratory condition may include reducing inflammation. A respiratory condition may be an inflammatory condition. In some embodiments, a respiratory condition may be or include airway disease, heart disease, vascular disease, or lung disease. In some embodiments, respiratory symptoms may be or be related to asthma, allergies, carbon monoxide poisoning, smoke inhalation, asbestos poisoning, bronchitis, pulmonary fibrosis, cystic fibrosis, embolism, chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome, pulmonary hypertension, Cesarean disease, or pneumonia. In some embodiments, the formulation containing AOM may be administered to a first tissue, which may be a deposition tissue or a target tissue. The formulation may be administered to the first tissue via inhalation. In some embodiments, such as those where the first tissue is not a target tissue, the formulation containing AOM may be applied to the first tissue and the formulation or formulation product (e.g., NO) may be delivered to a second tissue, for example, via diffusion. The second tissue may be a target tissue. In some embodiments, the depositing tissue, target tissue, or both may be associated with respiratory tissue. The depositing tissue, target tissue, or both may be the mucous membranes of an individual. The depositing tissue, target tissue, or both may be associated with the nose, nasopharynx, larynx, or trachea of ​​an individual. The target tissue may be the alveolar epithelium or lung parenchyma of an individual. The target tissue may be associated with the trachea (air duct), larynx, pharynx, small trachea, segmental bronchi, subsegmental bronchi, lung apex, pleura, pleural cavity, alveolar ducts, alveoli, main bronchus, lobar bronchi, hilum, upper lobe of the lung (including apical, posterior, and anterior segments), middle lobe of the lung (including medial basal and lateral segments), or lower lobe of the lung (including upper, posterior basal, or anterior basal segments). In some embodiments, the target tissue may be associated with the desired local effect. The desired local effect may involve, for example, treating infection, inflammation, or congestion. The desired local effect may involve, for example, treating at least one of the trachea, lungs, or other respiratory system tissues. In some embodiments, the desired local effect may be treating asthma, allergies, carbon monoxide poisoning, smoke inhalation, asbestos poisoning, bronchitis, pulmonary fibrosis, cystic fibrosis, embolism, chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome, pulmonary hypertension, or pneumonia. In some embodiments, the target tissue may be associated with the desired systemic effect. The desired systemic effect may include, for example, treatment of one or more of the following: headache, cardiovascular disease, inflammation, immune response, autoimmune disease, liver disease, infection, neurological disease, psychiatric disease, nitric oxide disorder, urea cycle disorder, congestion, vasodilatory disorder, skin disease, ophthalmic disease, wound healing, insect bite reaction, connective tissue disorder, and certain viral, bacterial, or fungal infections. In some embodiments, administration of an effective amount of an AOM-containing formulation may promote endothelial function. In some embodiments, administration of an effective amount of an AOM-containing formulation may alter or modify nitrite or NO levels in a target tissue or throughout the body. In some embodiments, administration of an effective amount of the formulation may modulate an individual's microbiome, such as the microbiome associated with an individual's respiratory system. In some embodiments, the administration of a formulation containing AOM may be device-assisted. In some embodiments, the formulation containing an AOM may be administered before the onset of a respiratory condition. In some embodiments, the formulation containing an AOM may be administered during the onset of a respiratory condition. In some embodiments, the formulation containing an AOM may be administered after the respiratory condition has subsided. In some embodiments, the formulation may be administered in response to respiratory condition symptoms, triggers, or warning signals. The formulation may be administered in response to cough or dyspnea. In some embodiments, the formulation may be administered before or after a surgical or diagnostic procedure. In some embodiments, the methods disclosed herein may further include determining whether an individual needs treatment for a respiratory condition. In some embodiments, formulations containing AOM may be administered in the form of a solution, suspension, or liquid. In some embodiments, formulations may be administered in the form of droplets, sprays, mists, or aerosols. In some embodiments, formulations may be formulated as a breathing solution (e.g., ultrafine droplets, aerosols, or mists), gas, or dry powder. Formulations may include microspheres or microcapsules. In some embodiments, the formulation may be formulated to be compatible with an individual's respiratory system. For example, the formulation may have a substantially physiological pH level. In some embodiments, the formulation may have a physiological volumetric osmotic concentration. For example, the formulation may be substantially isotonic. In some embodiments, formulations containing AOM may be formulated for immediate or prolonged release. In some embodiments, formulations containing AOM can be formulated for local or systemic delivery of nitrite or NO to target tissues. Formulations or their products can be formulated for transmucosal delivery and / or circulation, for example, locally or systemically. In some embodiments, the method disclosed herein may further include administering a second amount of an AOM-containing formulation to an individual. The formulation may be administered as part of a combination therapy. In some embodiments, the method may additionally include administration of a second treatment in combination with the formulation containing an AOM. The second treatment may include a surgical procedure. In some embodiments, the formulation containing an AOM may be administered in combination with an anti-inflammatory agent. The formulation may be administered in combination with a medical method approved for the treatment of, or commonly used for the treatment of, an associated disease or condition, or symptoms of an associated disease or condition. In some embodiments, the formulation containing AOM may be administered for a period of time prior to the initiation of the second treatment. In some embodiments, the formulation containing AOM may be administered concurrently with the second treatment. In some embodiments, the formulation containing AOM may be administered for a period of time after the cessation of the second treatment. In some embodiments, the second treatment may be administered via the respiratory system. The second treatment may be administered via alternating delivery modes, such as orally or intranasally. In some embodiments, an individual may have a second treatment at a therapeutic level. In some embodiments, formulations containing AOM may be administered in combination with asthma medications, cold and flu medications, corticosteroids, or antihistamines. Preparations containing AOM may be administered in combination with one or more therapeutic agents for asthma, allergies, carbon monoxide poisoning, asbestos poisoning, bronchitis, pulmonary fibrosis, cystic fibrosis, embolism, COPD, adult respiratory distress syndrome, pulmonary hypertension, or Cesarean disease. In some embodiments, preparations containing AOM may be administered in combination with nitrite, nitrate, and / or NO. In some embodiments, the effective amount of AOM or an AOM-containing formulation may be a therapeutically effective dose of AOM. The therapeutically effective dose of AOM may be about or greater than about 1×10³, 10⁴, 10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰, 10¹¹, 10¹², 10¹³, or 10¹⁴ CFU. In some embodiments, the formulation may be administered as an analgesic. In some embodiments, the formulation may be administered as a prophylactic agent. In some embodiments, the formulation may be self-administered. The method disclosed herein may involve administering a formulation containing AOM approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 times daily. The formulation may be administered approximately 1-3, 3-5, 5-7, 7-9, 5-10, 10-14, 12-18, 12-21, 21-28, 28-35, 35-42, 42-49, 49-56, 46-63, 63-70, 70-77, 77-84, or 84-91 days. In some embodiments, the formulation containing AOM may be administered to an individual within 30, 60, 90, 120, 150, or 180 minutes after waking from sleep. In some embodiments, the formulation containing AOM may be administered to an individual within 30, 60, 90, 120, 150, or 180 minutes before going to sleep. In some embodiments, the formulation may be administered to an individual within 30, 60, 90, 120, 150, or 180 minutes before eating. In some embodiments, the formulation may be administered to an individual within 30, 60, 90, 120, 150, or 180 minutes before cleaning or showering. In some embodiments, the individual may be female. In some embodiments, the individual may be male. The individual may be characterized as one of the following races / ethnicities: Asian, Black or African American, Hispanic or Latino, White, or multiracial. In some embodiments, the individual may be less than 1 year old, or between 1-5, 5-10, 10-20, 20-30, 30-40, 40-50, 50-60 years old, or over 60 years old. In some embodiments, the individual may have a disrupted microbiome. In some embodiments, the formulation may comprise a buffer solution containing AOM. The formulation may comprise an aqueous buffer solution containing AOM. The buffer solution may comprise disodium hydrogen phosphate and magnesium chloride. In some embodiments, the buffer solution may comprise water containing 50 mM Na₂HPO₄ and / or 2 mM MgCl₂. The buffer solution may consist essentially of disodium hydrogen phosphate and magnesium chloride, for example, essentially of water containing 50 mM Na₂HPO₄ and / or 2 mM MgCl₂. The buffer solution may consist of disodium hydrogen phosphate and magnesium chloride, for example, of water containing 50 mM Na₂HPO₄ and 2 mM MgCl₂. In some embodiments, the formulation containing AOM may contain at least one of ammonia, ammonium salts, and urea. In some embodiments, the formulation may further contain or be administered with a compound that promotes AOM growth or metabolism, NO production, and / or urease activity. In some embodiments, formulations containing AOM may include controlled-release materials. Formulations may also include slow-release materials. In some embodiments, the formulation may further comprise excipients. Formulations comprising AOM may comprise pharmaceutically acceptable excipients. Excipients may comprise one or more of the following: absorption and penetration enhancers, preservatives, antioxidants, buffers, chelating agents, ion exchangers, solubilizers, suspending agents, thickeners, surfactants, wetting agents, tension modifiers, enzyme inhibitors, and mediators for appropriate drug delivery. Mediators may be, for example, propellants. In some embodiments, excipients may comprise mucosal adhesives. In some embodiments, the formulation may comprise one or more of the following: disintegrants, chelating agents, coating agents, controlled-release products, or fillers. In some embodiments, formulations containing AOM may be substantially free of other organisms. Formulations containing AOM may further contain other organisms, such as biological communities. In some embodiments, the formulation containing AOM may contain between about 1×10³ CFU / mL and about 1×10¹⁴ CFU / mL of AOM. For example, the formulation may contain between about 1×10⁹ CFU / mL and about 10×10⁹ CFU / mL of AOM. In some embodiments, the AOM comprises ammonia-oxidizing bacteria (AOB). The AOM may consist essentially of AOB. In some embodiments, the AOM may comprise *Nitrobacter*, *Nitrobacter*, *Nitrobacterium*, *Nitrobacterium*, *Nitrobacterium*, *Nitrobacterium*, *Vibrio*, and combinations thereof. In some embodiments, the AOM may be *N. eutropha*. In some embodiments, the AOM may be *N. eutropha* D23 with ATCC registration number PTA-121157. In some embodiments, the AOM may comprise ammonia-oxidizing archaea (AOA). The AOM may consist essentially of AOA. In some embodiments, the AOM may be able to convert ammonia or ammonium to nitrite at a rate of at least about 1 pmol / min / mg protein. The AOM may also be able to convert ammonia or ammonium to nitrite at a rate of at least about 0.1 nanomol / min / mg protein. In some embodiments, the methods disclosed herein may include the use of a biocommunity-friendly product in combination with an AOM-containing formulation. According to one example, a formulation containing AOM, as disclosed herein, is provided to an individual via inhalation or intratracheal delivery. According to one example, preparations containing AOM as disclosed herein are provided for the treatment of individual respiratory conditions. In some embodiments, formulations containing an AOM may be packaged for single use. Formulations containing an AOM may be packaged for multiple uses. According to one embodiment, a device is provided that is structurally designed to deliver an AOM-containing formulation as disclosed herein. In some embodiments, the device is structurally designed to deliver an AOM-containing formulation to a target or deposited tissue of an individual's respiratory system. The device may be a nebulizer. The device may be a pressurized inhaler or a dry powder inhaler. The device may be an intratracheal device. According to one example, a kit containing an AOM as disclosed herein is provided. This invention covers all combinations of any one or more of the foregoing states and / or embodiments, as well as combinations with any one or more of the embodiments listed in the embodiments and any instances.

Implementation Method

Claims

1. Use of a preparation comprising non-pathogenic bacteria, wherein it is used to prepare a reagent for introducing non-pathogenic bacteria into an individual, wherein: The non-pathogenic bacteria are selected from species of the genera *Nitrosomonas*, *Nitrosococcus*, *Nitrosospira*, *Nitrosocystis*, *Nitrosolobus*, *Nitrosovibrio*, and combinations thereof, or *Streptococcus*. The preparation forms a plurality of particles with a particle size range (PSR) of about 5 micrometers or smaller. The plurality of particles are delivered to the individual's respiratory system, wherein the preparation is formulated for delivery to the individual via aerosol, inhalation, or intratracheal delivery.

2. Use of a preparation comprising non-pathogenic bacteria for the preparation of a pharmaceutical product for treating respiratory ailments in an individual, wherein: The non-pathogenic bacteria are selected from the genera *Nitrosomonas*, *Nitrosococcus*, *Nitrospirillum*, *Nitrostomyces*, *Nitrostomyces*, *Vibrio nitrosi*, and combinations thereof; the formulation forms a plurality of particles, the PSR of which is about 5 micrometers or smaller; and the plurality of particles are delivered to the respiratory system of the individual, wherein the formulation is prepared for delivery to the individual via aerosol, inhalation, or intratracheal delivery.

3. As requested in paragraph 1 or 2, wherein the non-pathogenic bacteria are Nitrosomonaseutropha (N. eutropha).

4. As requested in claim 3, wherein the non-pathogenic bacteria are eutrophic nitrosomonas D23 with ATCC accession number PTA-121157.

5. For the purposes of claim 2, wherein the respiratory condition is: (a) an inflammatory condition; (b) an airway disease; or (c) a vascular or pulmonary condition.

6. As requested in item 2, wherein the respiratory condition is asthma, allergy, carbon monoxide poisoning, smoke inhalation, emphysema, asbestos poisoning, bronchitis, pulmonary fibrosis, cystic fibrosis, embolism, chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome, pulmonary hypertension, Celic's disease, or pneumonia.

7. As claimed in claim 1 or 2, wherein at least 10% of each of the plurality of particles are live bacteria.

8. The use as claimed in claim 2, wherein the preparation is formulated as part of a combination therapy or wherein the preparation is formulated for combination with an anti-inflammatory agent.

9. As claimed in claim 1 or 2, wherein the preparation is formulated as a respiratory solution, gas or dry powder.

10. As requested in claim 9, wherein the breathing solution is an ultrafine droplet, aerosol, or atomized solution.

11. As claimed in claim 7, wherein at least 20% of each of the plurality of particles are live non-pathogenic bacteria.

12. As claimed in claim 7, wherein at least 30% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

13. As claimed in claim 7, wherein at least 40% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

14. As claimed in claim 7, wherein at least 50% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

15. As claimed in claim 7, wherein at least 60% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

16. As claimed in claim 7, wherein at least 70% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

17. As claimed in claim 7, wherein at least 80% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

18. As claimed in claim 7, wherein at least 85% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

19. As claimed in claim 7, wherein at least 90% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

20. As claimed in claim 7, wherein at least 95% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

21. As claimed in claim 7, wherein at least 99% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

22. As claimed in claim 7, wherein at least 99.9% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

23. As claimed in claim 7, wherein at least 99.99% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

24. As claimed in claim 7, wherein at least 99.999% of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

25. As requested in claim 7, wherein substantially all of the non-pathogenic bacteria in each of the plurality of particles are live bacteria.

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