WO2021211411A1 - Disinfectant compositions and methods of making and using the same - Google Patents

Abstract

Described herein are compositions comprising a halogen-containing compound and a second active agent, wherein the second active agent is selected from the group consisting of a metal, a chlorite, a chlorate, and combinations thereof. Methods of making and using such compositions are also described. Compositions of the present invention may be useful as a disinfectant.

Description

DISINFECTANT COMPOSITIONS AND METHODS OF MAKING AND

USING THE SAME

FIELD

[0001] The present invention relates to compositions comprising a halogen-containing compound and a second active agent, wherein the second active agent is selected from the group consisting of a metal, chlorite, chlorate, and combinations thereof. The present invention also relates to methods of making and using such compositions. Compositions of the present invention may be useful as a disinfectant.

BACKGROUND

[0002] Diarrheal disease is a leading cause of mortality for children under the age of five worldwide (Path, (2009). Diarrheal Disease: Solutions to Defeat a Global Killer. Path Publications, Washington) and was estimated to cause the deaths of approximately 500,000 children in this age group in 2015 (GBD Diarrhoeal Diseases Collaborators. The Lancet: Infectious Diseases, vol. 17, June 2016, pp. 909-948). In addition, infectious diseases represent health hazards to humans and animals of all ages across a range of global settings. Protozoan parasites such as Cryptosporidium species contribute to diarrheal diseases and are among the most frequent etiological causes of mortality among children, especially in developing nations (Omarova et al, (2018). International Journal of Environmental Research and Public Health, 75(3), 495). Parasitic protozoa such as Cryptosporidium species are microscopic single-celled obligate parasites capable of infecting humans and other mammals. Of all protozoan parasites, Cryptosporidium species are the most common etiological agents for waterborne disease outbreaks (Efstratiou, Artemis, et al. Water Research , vol. 114, May 2017, pp. 14-22). Chlorine-based disinfectants are highly effective for inactivating bacteria and viruses, and are among the most widely used methods for disinfecting drinking water worldwide. However, chlorine-based disinfectants have been reported to have very limited effectiveness with respect to inactivating Cryptosporidium parvum oocysts.

SUMMARY

[0003] A first aspect of the present invention is directed to a composition comprising: a halogen-containing compound (e.g., a halogen-containing isocyanurate such as sodium dichloroisocyanurate [commonly abbreviated NaDCC]); and a second active agent, wherein the second active agent is selected from the group consisting of a metal, chlorite, and chlorate.

[0004] A second aspect of the present invention is directed to a method of inactivating and/or killing a microorganism, the method comprising: exposing a microorganism to a composition of the present invention, thereby inactivating and/or killing the microorganism.

[0005] Another aspect of the present invention is directed to a method of inactivating and/or killing a microorganism, the method comprising: exposing a microorganism to a composition comprising a halogen-containing compound (e.g., a halogen-containing isocyanurate such as NaDCC) to form a mixture, wherein the mixture has a pH of less than about 6.5, thereby inactivating and/or killing the microorganism.

[0006] It is noted that aspects of the invention described with respect to one embodiment, may be incorporated in a different embodiment although not specifically described relative thereto. That is, all embodiments and/or features of any embodiment can be combined in any way and/or combination. Applicant reserves the right to change any originally filed claim and/or file any new claim accordingly, including the right to be able to amend any originally filed claim to depend from and/or incorporate any feature of any other claim or claims although not originally claimed in that manner. These and other objects and/or aspects of the present invention are explained in detail in the specification set forth below. Further features, advantages and details of the present invention will be appreciated by those of ordinary skill in the art from a reading of the figures and the detailed description of the preferred embodiments that follow, such description being merely illustrative of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS [0007] Fig. 1 illustrates a spot plate method for a disinfection assay of C. sporogenes spores on a RCA plate with serial dilutions at undiluted, (-1), and (-2). Sample spots were 10 pL each. UD = undiluted.

[0008] Fig. 2 illustrates a screw-capped vial containing water spiked with oocysts and disinfectant.

[0009] Fig. 3 is a flow chart describing a C. parvum cell culture- immunofluorescent microscopy infectivity assay.

[0010] Fig. 4 is an image of C. parvum infectious foci at 400X- immunofluorescent microscopy infectivity assay. DETAILED DESCRIPTION OF EXAMPLE EMBODIMENTS [0011] The present invention is now described more fully hereinafter with reference to the accompanying drawings, in which embodiments of the invention are shown. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather these embodiments are provided so that this disclosure will be thorough and complete and will fully convey the scope of the invention to those skilled in the art.

[0012] The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used in the description of the invention and the appended claims, the singular forms "a," "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0013] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the present application and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly so defined herein. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. In case of a conflict in terminology, the present specification is controlling.

[0014] Also as used herein, "and/or" refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative ("or").

[0015] Unless the context indicates otherwise, it is specifically intended that the various features of the invention described herein can be used in any combination. Moreover, the present invention also contemplates that in some embodiments of the invention, any feature or combination of features set forth herein can be excluded or omitted. To illustrate, if the specification states that a complex comprises components A, B and C, it is specifically intended that any of A, B or C, or a combination thereof, can be omitted and disclaimed. [0016] As used herein, the transitional phrase "consisting essentially of' (and grammatical variants) is to be interpreted as encompassing the recited materials or steps "and those that do not materially affect the basic and novel characteristic(s)" of the claimed invention. See, In re Herz, 537 F.2d 549, 551-52, 190 U.S.P.Q. 461, 463 (CCPA 1976) (emphasis in the original); see also MPEP § 2111.03. Thus, the term "consisting essentially of as used herein should not be interpreted as equivalent to "comprising."

[0017] The term "about," as used herein when referring to a measurable value such as an amount or concentration and the like, is meant to encompass variations of ± 10%, ± 5%, ± 1%, ± 0.5%, or even ± 0.1% of the specified value as well as the specified value. For example, "about X" where X is the measurable value, is meant to include X as well as variations of ± 10%, ± 5%, ± 1%, ± 0.5%, or even ± 0.1% of X. A range provided herein for a measureable value may include any other range and/or individual value therein.

[0018] As used herein, the terms "increase," "increases," "increased," "increasing," and similar terms indicate an elevation in the specified parameter or value of at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 300%, 400%, 500% or more.

[0019] As used herein, the terms "reduce," "reduces," "reduced," "reduction,"

"inhibit," and similar terms refer to a decrease in the specified parameter or value of at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, or 100%.

[0020] Provided according to embodiments of the present invention are compositions comprising a halogen-containing compound; and a second active agent, wherein the second active agent is selected from the group consisting of a metal, a chlorite, a chlorate, and combinations thereof. A composition of the present invention may be used as a disinfectant (e.g., a chemical disinfectant). In some embodiments, a composition of the present invention may be used to disinfect water (e.g., drinking water), food, beverages, surfaces, compositions and/or articles of manufacture including surfaces thereof. A composition of the present invention may be in the form of a solid (e.g., powder, tablet, etc.) or in the form of a liquid. In some embodiments, a composition of the present invention is in the form of a sachet, capsule, and/or pod, optionally a dispersible sachet, capsule, and/or pod.

[0021] As described herein, in some embodiments, a composition of the present invention may be contacted (e.g., added to and/or combined with) to another composition such as a liquid (e.g., water) to form a mixture. Accordingly, a composition of the present invention may be formulated to have certain concentrations and/or properties upon contact (e.g., combination) with another composition such as a liquid (e.g., water). For example, in some embodiments, a composition of the present invention is in the form of a solid (e.g., tablet, granules, etc.), and the solid dissolves upon contact (e.g., combination) with a liquid (e.g., water), optionally at a temperature in a range from about 15°C or 20°C to about 22°C or 25°C and/or in about 1, 2, 5, 10, or 15 minute(s) to about 20, 25, 30, 35, 40, 45, 50, 55 or 60 minutes after contact and/or combination of the composition and the liquid. In some embodiments, a composition of the present invention is in the form of a solid (e.g., tablet, granules, etc.), and the solid dissolves upon contact (e.g., combination) with a liquid (e.g., water), optionally at room temperature (e.g., about 20°C to about 22°C) and/or in about 1, 2, 5, 10, or 15 minute(s) to about 20, 25, 30, 35, 40, 45, 50, 55 or 60 minutes after contact and/or combination of the composition and the liquid. In some embodiments, a composition of the present invention is in the form of a tablet and the tablet dissolves upon contact with a liquid (e.g., water), optionally at room temperature (e.g., about 20°C to about 22°C) and/or in about 1, 2, 5, 10, or 15 minute(s) to about 20, 25, 30, 35, 40, 45, 50, 55 or 60 minutes after contact of the tablet and the liquid. The tablet may be configured to dissolve and/or dissolves in about 1 liter, 5 liters, 10 liters, and/or 20 liters of a liquid (e.g., water) and/or may be configured to dissolve and/or dissolves in a 1 liter, 5 liter, 10 liter, and/or 20 liter container comprising a liquid (e.g., water), such as, in an amount of about 1 liter, 5 liters, 10 liters, and/or 20 liters, respectively. In some embodiments, the 1 liter, 5 liter, 10 liter, and/or 20 liter container comprises the liquid (e.g., water) in an amount that provides less than about 5% headspace in the container. In some embodiments, a composition of the present invention (e.g., one of more (e.g., 1, 2, 3, 4, or more) tablet(s)) may be formulated to have a desired concentration of one or more component(s) (e.g., ingredients such as a halogen-containing compound and/or second active agent) upon contact and/or dissolution in a given volume of water (e.g., in about 1, 5, or 10 liter(s) to about 20, 25, 30, 35, 40, 45, or 50 liters such as about 1 L, 5 L, 10 L, or 20 L of water). For example, in some embodiments, a composition of the present invention is one tablet or two tablets that dissolve in about 20 liters of a liquid (e.g., water) and/or that dissolve in a 20 liter container comprising a liquid (e.g., water) in an amount of about 20 liters and/or in an amount that provides less than about 5% headspace in the container to thereby provide a desired concentration of one or more component(s) of the composition in the liquid.

[0022] In some embodiments, a composition of the present invention has no or minimal detectable taste and/or odor, optionally upon combination with a liquid (e.g., water, beverage, etc.) and/or food. The composition may include one or more food-grade and/or USP-grade ingredient(s). In some embodiments, the composition includes only food-grade and/or USP-grade ingredients. [0023] A halogen-containing compound of the present invention is a compound that includes a halogen (e.g., chlorine, bromine, and/or iodine). A halogen containing compound may provide a source of free chlorine, a source of combined chlorine such as chloramines or other organic compounds containing chlorine and having the potential to act as an oxidant, a source of monochloramine, a source of dichloramine, a source of other chloramines (e.g., N- chloramines, chloramine-T and/or chloramine-B), a source of chlorine dioxide, a source of bromine, a source of iodine, and/or a source of brominated and/or iodinated halamines. "Free chlorine" as used herein refers to Ch, HOC1, OC1 , and/or another source of chlorine or hypochlorite that not combined with an organic compound. "Combined chlorine" as used herein refers to chloramines and other reactive species in which chlorine is combined with an organic compound. Exemplary halogen-containing compounds of the present invention include, but are not limited to, chlorine-containing compounds such as a dichloroisocyanurate (e.g., sodium dichloroisocyanurate (NaDCC)) and/or a trichloroisocyanurate (e.g., sodium trichloroisocyanurate); bromine-containing compounds such as a dibromooisocyanurate (e.g., sodium dibromoisocyanurate) and/or a tribromoisocyanurate (e.g., sodium tribromoisocyanurate); and/or an iodine-containing compound such as a diiodoisocyanurate (e.g., sodium diiodoisocyanurate) and/or a triiodomoisocyanurate (e.g., sodium triiodoisocyanurate). A halogen-containing compound (e.g., NaDCC) may be present in a composition of the present invention in any suitable form. In some embodiments, the halogen-containing compound is present in the composition in an anhydrous form and/or a hydrated form (e.g., in the form of a dihydrate or other hydrate form).

[0024] In some embodiments, a composition of the present invention comprises a dichloroisocyanurate. In some embodiments, a composition of the present invention comprises NaDCC. In some embodiments, a composition of the present invention comprises dichloroisocyanurate (e.g., NaDCC) and at least one additional halogen containing compound (e.g., a different chlorine-containing compound, a bromine-containing compound, and/or an iodine-containing compound).

[0025] One or more (e.g., 1, 2, 3, or more) halogen-containing compound(s) may each be present in the composition in an amount of about 0.001% or 0.01% to about 10% or 25% by weight of the composition. In some embodiments, a halogen-containing compound (e.g., NaDCC) may be present in the composition in an amount of about 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, or 25% by weight of the composition. In some embodiments, a halogen-containing compound (e.g., NaDCC) may be present in the composition in an amount of about 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, or 2.5% to about 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, or 25% by weight of the composition. In some embodiments, one or more halogen-containing compound(s) may each be present in the composition in an amount of about 0.001, 0.01, or 0.1 mg/L to about 10 or 100 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, one or more halogen-containing compound(s) may each have a concentration of about 0.001, 0.01, or 0.1 mg/L to about 10 or 100 mg/L in the mixture. In some embodiments, a halogen-containing compound (e.g., NaDCC) may be present in the composition in an amount of about 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 2, 3, or 4 mg/L to about 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, one or more halogen- containing compound(s) may each have a concentration of about 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 2, 3, or 4 mg/L to about 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 mg/L in the mixture. In some embodiments, one or more halogen-containing compound(s) are present in the composition in a biocidal amount. In some embodiments, NaDCC is present in a composition of the present invention in a biocidal amount.

[0026] One or more metal(s) (e.g., 1, 2, 3, 4, 5 or more) may each be present in a composition of the present invention. Exemplary metals include, but are not limited to, copper, silver, gold, iron, manganese, and/or zinc. A metal may be present in a composition of the present invention in the form of a metal salt, a metal powder, a metal oxide, a metal oxyhydroxide, a metal hydrate, an aqueous metal solution, and/or a metal chelate. A metal may have a size configured to dissolve and/or disperse in a liquid (e.g., water), optionally upon exposure and/or contact with the liquid. In some embodiments, a metal present in a composition of the present invention may have a diameter of about 1, 5, 10, 15, 20, 25, 30, 35, 40, or 45 microns to about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 microns. In some embodiments, a metal powder present in a composition of the present invention may have a particle size of about 20, 30, 40, 50, 60, 70, 80, 90, 100, 140, 200, 230, 275, 300, or 325 mesh to about 400, 625, 1250, or 2500 mesh.

[0027] One or more metals may each be present in the composition in an amount of about 0.001% to about 10% by weight of the composition. In some embodiments, a metal is present in the composition in an amount of about 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% by weight of the composition. In some embodiments, a metal may be present in the composition in an amount of about 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, or 2.5% to about 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% by weight of the composition. In some embodiments, one or more metal(s) may each be present in the composition in an amount of about 0.0001 mg/L to about 100 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, one or more metal(s) may each have a concentration of about 0.0001 mg/L to about 100 mg/L in the mixture. In some embodiments, a metal may be present in the composition in an amount of about 0.0001, 0.005, 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, or 45 mg/L to about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, one or more metal(s) may each have a concentration of about 0.0001, 0.005, 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, or 45 mg/L to about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 mg/L in the mixture. In some embodiments, one or more metals(s) are present in the composition in a biocidal amount.

[0028] In some embodiments, a composition of the present invention comprises a halogen-containing compound (e.g., NaDCC) and at least one metal. The metal and halogen- containing compound may be present in the composition in a ratio, by weight, of about 1:10000 or 1:1000 to about 2:1 or 10:1 (metal : halogen-containing compound) such as about 1:10000, 1:5000, 1:1000, 1:750, 1:500, 1:250, 1:100, 1:50, 1:10, 1:2, 1:1, 2:1, 5:1, or 10:1. In some embodiments, the composition comprises copper, silver and/or zinc.

[0029] In some embodiments, a composition of the present invention comprises copper. Copper may be present in the composition in an amount of about 0.0001% to about 10% by weight of the composition. In some embodiments, copper is present in the composition in an amount of about 0.0001%, 0.0005%, 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% by weight of the composition. In some embodiments, copper is present in the composition in an amount of about 0.0001%, 0.0005%, 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, or 2.5% to about 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% by weight of the composition. In some embodiments, copper may be present in the composition in an amount of about 0.01 or 0.1 mg/L to about 10 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, copper is present in the mixture in an amount of about 0.01 or 0.1 mg/L to about 10 mg/L. In some embodiments, copper may be present in the composition in an amount of about 0.01, 0.05, 0.1, 0.5, 1, 2, 3, or 4 mg/L to about 5, 6, 7, 8, 9, or 10 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, copper is present in the mixture in an amount of 0.01, 0.05, 0.1, 0.5, 1, 2, 3, or 4 mg/L to about 5, 6, 7, 8, 9, or 10 mg/L. In some embodiments, a composition of the present invention comprises copper and a halogen-containing compound (e.g., NaDCC) in a ratio, by weight, of about 1:10000 or 1:1000 to about 2:1 or 10:1 (copper : halogen-containing compound) such as about 1:10000, 1:5000, 1:1000, 1:750, 1:500, 1:250, 1:100, 1:50, 1:10, 1:2, 1:1, 2:1, 5:1 or 10:1. In some embodiments, copper may be present in the composition in a biocidal amount. [0030] In some embodiments, a composition of the present invention comprises silver. Silver may be present in the composition in an amount of about 0.0001% to about 1% or 10% by weight of the composition. In some embodiments, silver is present in the composition in an amount of about 0.0001%, 0.0005%, 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% by weight of the composition. In some embodiments, silver is present in the composition in an amount of about 0.0001%, 0.0005%, 0.001%, 0.005%, 0.01%, or 0.05% to about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% by weight of the composition. In some embodiments, silver may be present in the composition in an amount of about 0.00001 or 0.001 mg/L to about 1 or 2 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, silver is present in the mixture in an amount of about 0.00001 or 0.001 mg/L to about lor 2 mg/L. In some embodiments, silver may be present in the composition in an amount of about 0.00001, 0.00005, 0.0001, 0.0005, 0.001, 0.005, 0.01, 0.05, or 0.1 to about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, or 2 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, silver is present in the mixture in an amount of about 0.00001, 0.00005, 0.0001, 0.0005, 0.001, 0.005, 0.01, 0.05, or 0.1 to about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, or 2 mg/L. In some embodiments, a composition of the present invention comprises silver and a halogen-containing compound (e.g., NaDCC) in a ratio, by weight, of about 1:10000 to about 1:1 or 10:1 (silver : halogen- containing compound) such as about 1:10000, 1:5000, 1:1000, 1:750, 1:500, 1:250, 1:100, 1:50, 1:10, 1:2, 1:1, 2:1, 5:1, or 10:1. In some embodiments, silver may be present in the composition in a biocidal amount.

[0031] In some embodiments, a composition of the present invention comprises zinc.

Zinc may be present in the composition in an amount of about 0.001% or 0.01% to about 10% by weight of the composition. In some embodiments, zinc is present in the composition in an amount of about 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% by weight of the composition. In some embodiments, zinc is present in the composition in an amount of about 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, or 2.5% to about 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% by weight of the composition. In some embodiments, zinc may be present in the composition in an amount of about 0.01 or 1 mg/L to about 1000 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, zinc is present in the mixture in an amount of about 0.01 or 1 mg/L to about 1000 mg/L. In some embodiments, zinc may be present in the composition in an amount of about 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 50, or 100 mg/L to about 150, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, zinc is present in the mixture in an amount of about 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 50, or 100 mg/L to about 150, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 mg/L. In some embodiments, a composition of the present invention comprises zinc and a halogen-containing compound (e.g., NaDCC) in a ratio, by weight, of about 1:1000 or 1:100 to about 100:1 (zinc : halogen-containing compound) such as about 1:1000, 1:500, 1:100, 1:75, 1:50, 1:25, 1:10, 1:5, 1:1, 5:1, 10:1, 25:1, 50:1, 75:1, or 100:1. In some embodiments, zinc may be present in the composition in a biocidal amount.

[0032] A composition of the present invention may comprise a halogen-containing compound (e.g., NaDCC), copper, silver, and zinc.

[0033] In some embodiments, a composition of the present invention comprises a halogen-containing compound (e.g., NaDCC) and at least one of a chlorite and/or a chlorate. Exemplary chlorites include, but are not limited to, sodium chlorite and/or potassium chlorite. Exemplary chlorates include, but are not limited to, sodium chlorate and/or potassium chlorate. In some embodiments, a composition of the present invention comprises a halogen- containing compound (e.g., NaDCC), a least one metal (e.g., silver, zinc, and/or copper) and a chlorite and/or a chlorate. In some embodiments, a composition of the present invention comprises a halogen-containing compound (e.g., NaDCC), a least one metal (e.g., silver, zinc, and/or copper) and a chlorite. In some embodiments, a composition of the present invention comprises a halogen-containing compound (e.g., NaDCC), a least one metal (e.g., silver, zinc, and/or copper) and a chlorate. In some embodiments, a composition of the present invention comprises a halogen-containing compound (e.g., NaDCC), a least one metal (e.g., silver, zinc, and/or copper), a chlorite and a chlorate.

[0034] One or more (e.g., 1, 2, 3, or more) of a chlorite and/or chlorate may each be present in the composition in an amount of about 0.001% to about 10% by weight of the composition. In some embodiments, a chlorite and/or chlorate is present in the composition in an amount of about 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% by weight of the composition. In some embodiments, a chlorite and/or chlorate may be present in the composition in an amount of about 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, or 2.5% to about 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% by weight of the composition. In some embodiments, one or more of a chlorite and/or chlorate may each be present in the composition in an amount of about 0.001 or 0.1 mg/L to about 1 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, the chlorite and/or chlorate is present in the mixture in an amount of about 0.001 or 0.1 mg/L to about 1 mg/L. In some embodiments, a chlorite and/or chlorate may be present in the composition in an amount of about 0.001, 0.005, 0.1, 0.2, 0.3, or 0.4 mg/L to about 0.5, 0.6, 0.7, 0.8, 0.9, or 1 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, the chlorite and/or chlorate is present in the mixture in an amount of about 0.001, 0.005, 0.1, 0.2, 0.3, or 0.4 mg/L to about 0.5, 0.6, 0.7, 0.8, 0.9, or 1 mg/L. In some embodiments, a composition of the present invention comprises a chlorite and a halogen-containing compound (e.g., NaDCC) in a ratio, by weight, of about 1:1000 to about 1:1 (chlorite: halogen-containing compound) such as about 1:1000, 1:750, 1:500, 1:250, 1:100, 1:50, 1:10, 1:2, or 1:1. In some embodiments, a composition of the present invention comprises a chlorate and a halogen-containing compound (e.g., NaDCC) in a ratio, by weight, of about 1:1000 to about 1:1 (chlorate: halogen-containing compound) such as about 1:1000, 1:750, 1:500, 1:250, 1:100, 1:50, 1:10, 1:2, or 1:1. In some embodiments, a chlorite and/or a chlorate are present in the composition in a biocidal amount. [0035] A composition of the present invention may comprise a coagulant. A coagulant may be present in a composition of the present invention that is used in emergency settings and/or in turbid waters. Exemplary coagulants include, but are not limited to, a chitosan such as chitosan acetate, chitosan lactate, and chitosan HC1. The coagulant may be nontoxic and/or biodegradable.

[0036] One or more (e.g., 1, 2, 3, or more) coagulant(s) may each be present in the composition in an amount of about 0.001% to about 10% by weight of the composition. In some embodiments, a coagulant (e.g., a chitosan) is present in the composition in an amount of about 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% by weight of the composition. In some embodiments, a coagulant is present in the composition in an amount of about 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, or 2.5% to about 3%, 3.5%, 4% 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% by weight of the composition. In some embodiments, a coagulant may be present in the composition in an amount of about 0.01 or 0.1 mg/L to about 100 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, a coagulant is present in the mixture in an amount of about 0.01 or 0.1 mg/L to about 100 mg/L. In some embodiments, a coagulant may be present in the composition in an amount of about 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, or 45 mg/L to about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 mg/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, a coagulant is present in the mixture in an amount of about 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, or 45 mg/L to about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 mg/L. In some embodiments, a composition of the present invention comprises a coagulant and a halogen-containing compound (e.g., NaDCC) in a ratio, by weight, of about 1:1000 to about 100:1 (coagulant: halogen-containing compound) such as about 1:1000, 1:750, 1:500, 1:250, 1:100, 1:50, 1:10, 1:2, 1:1, 2:1, 10:1, 50:1, or 100:1.

[0037] A carbonate may be present in a composition of the present invention.

Exemplary carbonates include, but are not limited to, sodium hydrogen carbonate, sodium carbonate, sodium sesquicarbonate, potassium hydrogen carbonate, potassium carbonate, potassium sesquicarbonate, ammonium hydrogen carbonate, ammonium carbonate, ammonium sesquicarbonate, and/or bicarbonates (e.g., sodium bicarbonate).

[0038] One or more (e.g., 1, 2, 3, 4, or more) carbonate(s) may each be present in the composition in an amount of about 1% to about 95% by weight of the composition. In some embodiments, a carbonate is present in the composition in an amount of about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% by weight of the composition. In some embodiments, a carbonate is present in the composition in an amount of about 1% to about 5% or 10%; about 1%, 5%, or 10% to about 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%; about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% to about 80%, 85%, 90%, or 95%; or 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% to about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% by weight of the composition. In some embodiments, a carbonate (e.g., sodium carbonate) is present in the composition in an amount of about 1% to about 5% or 10% by weight of the composition. In some embodiments, a bicarbonate (e.g., sodium bicarbonate) is present in the composition in an amount of about 1%, 5%, or 10% to about 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% by weight of the composition.

[0039] In some embodiments, a carbonate may be present in the composition in an amount of about 0.001 g/L to about 10 g/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, a carbonate is present in the mixture in an amount of about 0.001 g/L to about 10 g/L. In some embodiments, a carbonate may be present in the composition in an amount of about 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, or 2 g/L to about 3, 4, 5, 6, 7, 8, 9, or 10 g/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, a carbonate is present in the mixture in an amount of about 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, or 2 g/L to about 3, 4, 5, 6, 7, 8, 9, or 10 g/L.

[0040] One or more (e.g., 1, 2, 3, or more) buffering agent(s) may each be present in a composition of the present invention. A “buffering agent” as used herein is an agent that is used to modify the pH of the composition. Exemplary buffering agents are acids and bases. Exemplary bases include, but are not limited to, sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, and/or combinations thereof. Exemplary acids include, but are not limited to, organic acids such as citric acid, adipic acid, tartaric acid, malic acid, fumaric acid, and phosphoric acid; acidic salts of organic acids; straight-chain aliphatic acids such as acetic acid, propanoic acid, butyric acid and valeric acid; dicarboxylic acids such as oxalic acid, malonic acid, and succinic acid; acidic amino acids such as glutamic acid and aspartic acid; hydroxy acids such as glycolic acid, lactic acid, and a-hydroxy butyric acid; and inorganic acids such as phosphoric acid, potassium dihydrogen phosphate, sodium dihydrogen phosphate, sodium sulfite, sodium bisulfate, potassium sulfite, potassium bisulfate, and sulfamic acid; and any combination thereof.

[0041] One or more (e.g., 1, 2, 3, 4, or more) buffering agent(s) may each be present in the composition in an amount of about 0.01% to about 95% by weight of the composition. In some embodiments, a buffering agent is present in the composition in an amount of about 0.01%, 0.05%, 0.1%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% by weight of the composition. In some embodiments, a buffering agent is present in the composition in an amount of about 0.01% to about 0.5%; about 0.01% to about 50%; about 0.1% to about 10%; about 1% to about 5% or 10%; about 1%, 5%, or 10% to about 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%; about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% to about 80%, 85%, 90%, or 95%; or 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% to about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% by weight of the composition. In some embodiments, an acid (e.g., adipic acid) is present in the composition in an amount of about 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% to about 80%, 85%, 90%, or 95% by weight of the composition.

[0042] In some embodiments, a buffering agent (e.g., an acid) may be present in the composition in an amount of about 0.01 g/L to about 100 g/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, a buffering agent is present in the mixture in an amount of about 0.01 g/L to about 100 g/L. In some embodiments, a buffering agent may be present in the composition in an amount of about 0.01, 0.05, 0.1, 0.5, 1, 5, 10, or 15 g/L to about 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, or 100 g/L and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, a buffering agent is present in the mixture in an amount of about 0.01, 0.05, 0.1, 0.5, 1, 5, 10, or 15 g/L to about 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, or 100 g/L.

[0043] A composition of the present invention may have a pH of about 2, 2.5, 3, 3.5,

4, 4.5, 5, or 5.5 to about 6, 6.5, or 7 and/or, when a composition of the present invention is combined with a liquid (e.g., water) to form a mixture, the mixture has a pH of about 2, 2.5, 3, 3.5, 4, 4.5, 5, or 5.5 to about 6, 6.5, or 7. In some embodiments, a composition of the present invention has a pH of about 6.5 or less and/or, when a composition of the present invention is combined with a liquid (e.g., water) to form a mixture, the mixture has a pH of about 6.5 or less. A buffering agent may be present in a composition of the present invention to provide a pH of about 2, 2.5, 3, 3.5, 4, 4.5, 5, or 5.5 to about 6, 6.5, or 7 and/or a buffering agent may be present in the composition such that when the composition is combined with a liquid (e.g., water) to form a mixture, the mixture has a pH of about 2, 2.5, 3, 3.5, 4, 4.5, 5, or 5.5 to about 6, 6.5, or 7. In some embodiments, a buffering agent may be present in a composition of the present invention to provide a pH of less than about 6.5 and/or a buffering agent may be present in the composition such that when the composition is combined with a liquid (e.g., water) to form a mixture, the mixture has a pH of less than about 6.5.

[0044] One or more components (e.g., a carbonate and an acid) may each be present in a composition of the present invention and provide the composition with a buffer strength of about 0.1 mM to about 10 M, optionally about 0.1 mM to about 1 M, about 1 mM to about 1 M, about 10 mM to 1 M, or about 100 mM to about 1 M. In some embodiments, a composition of the present invention comprises a carbonate buffer, a bisulfate buffer (e.g., includes sodium bisulfate), a phosphate buffer, and/or a citric acid buffer. In some embodiments, an acid (e.g., adipic acid) and a carbonate and/or a bicarbonate are present in the composition and provide the composition with a buffer strength of about 0.1 mM to about 10 M.

[0045] In some embodiments, one or more components in a composition of the present invention may provide effervescence. Exemplary effervescent agents include, but are not limited to, a carbonate, bicarbonate and/or an acid (e.g., a dry acid (i.e., a solid that dissolves in water to provide an acidic solution such as granular sodium bisulfate). In some embodiments, the acid is adipic acid. A composition of the present invention may comprise a carbonate and an acid and combination of the composition with an aqueous solution (e.g., water) may provide and/or result in effervescence.

[0046] One or more (e.g., 1, 2, 3, or more) additive(s) may each be present in a composition of the present invention. Exemplary additives include, but are not limited to, a surfactant, a source of peroxide (e.g., a percarbonate), a source of peracetic acid (e.g., sodium percarbonate and/or tetraacetylethylenediamine (TAED)), and/or performic acid (e.g., sodium percarbonate and/or formic acid in the presence of an acid catalyst such as sulfuric acid or sodium bisulfate, etc.). In some embodiments, an additive (e.g., peracetic acid and/or peformic acid) is present in a composition of the present invention at a concentration of about 0.1 mg/L to about 100,000 mg/L such as about 0.1, 1, 10, 100, or 1,000 mg/L to about 5,000, 10,000, 50,000, or 100,000 mg/L. In some embodiments, when a composition of the present invention is combined with a liquid (e.g., water) to form a mixture, an additive (e.g., peracetic acid and/or peformic acid) is present in the mixture in an amount of about 0.1 mg/L to about 100,000 mg/L such as about 0.1, 1, 10, 100, or 1,000 mg/L to about 5,000, 10,000, 50,000, or 100,000 mg/L.

[0047] A composition of the present invention may provide a biocidal concentration of a halogen and/or a halogen-containing compound. A "biocidal concentration" or “biocidal amount” as used herein refer to a concentration or amount of a compound or element that is capable of inactivating and/or killing a microorganism. "Inactivating," "inactivates," and grammatical variations thereof as used herein refer to preventing, disrupting, and/or reducing the ability of a microorganism to replicate, grow, and/or survive and/or to preventing, disrupting, and/or reducing the infectivity, metabolic function, and/or genetic integrity of a microorganism. In some embodiments, a composition of the present invention has and/or provides a biocidal concentration of chlorine, optionally wherein chlorine may be present in the composition and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, in the mixture as hypochlorous acid, hypochlorite, and/or a combined chlorine (e.g., monochloramine, dichloramine, etc.). In some embodiments, a composition of the present invention has and/or provides a biocidal concentration of a bromine, optionally wherein the bromine may be present in the composition and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, in the mixture as hypobromous acid, hypobromite, and/or a combined bromine (e.g., monobromamine, dibromamine, etc.). In some embodiments, a composition of the present invention has and/or provides a biocidal concentration of an iodine, optionally wherein the iodine may be present in the composition and/or, when the composition is combined with a liquid (e.g., water) to form a mixture, in the mixture as hypoiodous acid, hypoiodite, and/or a combined iodine. In some embodiments, a composition of the present invention has and/or provides a biocidal concentration of chlorine dioxide. In some embodiments, hypochlorous acid (HOC1) and/or free chlorine are present in a composition of the present invention at a concentration of about 5 mg/L or less, optionally of about 0.1, 0.5, 1, 1.5, 2, or 2.5 mg/L to about 3, 3.5, 4, 4.5, or 5 mg/L. In some embodiments, when a composition of the present invention is combined with a liquid (e.g., water) to form a mixture, hypochlorous acid (HOC1) and/or free chlorine is present in the mixture in an amount of about 5 mg/L or less, optionally of about 0.1, 0.5, 1, 1.5, 2, or 2.5 mg/L to about 3, 3.5, 4, 4.5, or 5 mg/L.

[0048] A composition of the present invention may provide a continuous release of a halogen, halogen-containing compound, and/or biocidal compound for a period of time (e.g., for about 1, 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 minutes or more or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 20, or 24 hours or more), optionally wherein the composition provides a continuous release of chlorine, chlorine dioxide, HOC1, bromine, and/or iodine for a period of time. The continuous release of the halogen, halogen-containing compound, and/or biocidal compound may be during and/or after exposure and/or combination of the composition with a liquid (e.g., water), optionally to form a mixture. In some embodiments, a composition of the present invention provides a release of a halogen, halogen-containing compound, and/or biocidal compound in an amount sufficient to inactivate and/or kill a microorganism exposed to and/or in contact with the composition and/or for a period of time sufficient to inactivate and/or kill a microorganism exposed to and/or in contact with the composition. A composition of the present invention may provide a release of a halogen, halogen-containing compound, and/or biocidal compound for about 1, 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 minutes or more or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 20, or 24 hours or more.

[0049] In some embodiments, a composition of the present invention provides a synergistic effect. In some embodiments, combination of a halogen-containing compound (e.g., NaDCC) and a second active agent (e.g., a metal, chlorite, and/or chlorate) in a composition of the present invention provides a synergist effect. "Synergistic", "synergy", or grammatical variants thereof as used herein refer to a combination exhibiting an effect greater than the effect that would be expected from the sum of the effects of the individual active ingredients of the combination alone. For example, the terms "synergistic" or "synergy" with regard to a composition comprising a halogen-containing compound (e.g., NaDCC) and a second active agent (e.g., a metal, chlorite, and/or chlorate) refers to an effect (e.g., inactivation and/or killing of a microorganism) that is greater than that which would be expected from the sum of the individual effects of the halogen-containing compound (e.g., NaDCC) and the second active agent (e.g., a metal, chlorite, and chlorate) alone.

[0050] According to some embodiments, a composition of the present invention may be used to inactivate and/or kill a microorganism present on and/or in a material and/or composition. Exemplary microorganisms include, but are not limited to, protozoans, protozoan cysts, helminths, helminth eggs, bacteria, spores, fungi, and/or viruses such as coronaviruses (e.g., severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and/or SARS-CoV-2). In some embodiments, the microorganism is Giardia, a Cryptosporidium species (e.g., Cryptosporidium parvum ) and/or oocysts thereof, and/or Clostridium species and/or spores thereof. In some embodiments, a method of the present invention inactivates and/or kills Cryptosporidium parvum and/or Cryptosporidium parvum oocysts.

[0051] A composition and/or method of the present invention may inactivate and/or kill a microorganism (e.g., a Cryptosporidium species, Clostridium species, and/or oocysts thereof) at a rate in which a log reduction value of about 1, 2, 3, 4, 5, 6, or more in the number of microorganisms is achieved at about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, or 55 minutes, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 hour(s) or more after the microorganism is exposed to the composition, optionally as measured by cell culture followed by immunofluorescent microscopy and/or PCR. In some embodiments, a composition and/or method of the present invention has about a 2-log or greater reduction of a microorganism in about 8 hours after the microorganism is exposed to the composition. In some embodiments, a composition and/or method of the present invention has about a 3 -log or greater reduction of a microorganism in about 2 hours or less after the composition is exposed to the microorganism. "Exposing," "exposed," "exposure," and grammatical variants thereof as used herein refer to placing a composition and microorganism under conditions suitable for the microorganism to come into contact with the composition and/or a component thereof (e.g., a component present in a biocidal amount) to achieve the desired inactivation of the microorganism and/or to kill the microorganism. Thus, for example, exposing a microorganism to a composition of the present invention may comprise contacting the microorganism with the composition (e.g., adding, pouring and/or the like the composition onto the microorganism and/or into or onto a medium such as water or a surface in and/or on which the microorganism is present). In some embodiments, exposing a microorganism to a composition of the present invention may comprise combining the composition with the microorganism and/or with a medium or surface in and/or on which the microorganism is present such as by adding the composition to the microorganism or vice versa.

[0052] In some embodiments, a composition of the present invention may be used to inactivate and/or kill a microorganism present on a surface of a composition and/or substrate (e.g., a substrate such as a desk, table, bench, bed, container, fabric, medical device and/or equipment, etc.) and/or in a composition and/or substrate (e.g., fabric, sponge, wastewater, excreta, soil, etc.). A microorganism may be present in and/or on a composition (e.g., a liquid such as water) and exposing the microorganism to a composition of the present invention may comprise combining the two compositions in any manner, optionally to form a mixture. In some embodiments, a microorganism may be present in and/or on an article of manufacture (e.g., desk, container, fabric, table, bench, bed, medical device and/or equipment, etc.) and exposing the microorganism to a composition of the present invention may comprise contacting the composition to the article of manufacture in any manner, optionally spraying, pouring, submerging, washing, and/or the like. Prior to exposure to a composition of the present invention, one or more additional component(s) may be present with a microorganism such as water, dirt, etc. such as when the microorganism is present in and/or on a composition and/or in and/or on an article of manufacture.

[0053] According to some embodiments, a method of inactivating and/or killing a microorganism comprises exposing a microorganism to a composition of the present invention, thereby inactivating and/or killing the microorganism. In some embodiments, the composition of the present invention has a pH of less than about 6.5. In some embodiments, exposing the microorganism to the composition comprises forming a mixture that has a pH of less than about 6.5. [0054] In some embodiments, exposing the microorganism to the composition is performed and/or carried out in a container comprising the composition and microorganism, optionally with one or more additional components (e.g., water), and the container has less than about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, or 1% headspace. In some embodiments, the container has less than about 5% headspace. In some embodiments, the container has minimal headspace or no headspace. "Headspace" as used herein refers to the space occupied by air in the container.

[0055] A composition of the present invention may be used to disinfect a liquid (e.g., water, a beverage, etc.) and/or food by inactivating and/or killing a microorganism. In some embodiments, a composition and/or method of the present invention is used to disinfect water and/or to provide drinking water for use in household water treatment and safe storage (HWTS) applications, optionally in emergency and/or low- and middle-income country (LMIC) settings. In some embodiments, a composition of the present invention may provide water suitable for drinking and/or animal (e.g., human, livestock, etc.) consumption. In some embodiments, a composition of the present invention may be used to disinfect a surface (e.g., a surface of an article of manufacture) by inactivating and/or killing a microorganism.

[0056] The present invention is explained in greater detail in the following non limiting Examples.

EXAMPLES

Example 1:

[0057] Disinfectant formulations were developed and tested to quantify their performance with respect to the inactivation of Cryptosporidium parvum oocysts in water and their potential to be granted regulatory approval for drinking water disinfection. This work was conducted in several phases. Rapid high-throughput screening of candidate disinfectants (in liquid solution) was conducted using Clostridium sporogenes spores as surrogates for C. parvum. Candidate disinfectants that efficiently inactivated C. sporogenes spores (> 2-log inactivation in 8 hours) were evaluated with respect to their ability to achieve substantive inactivation of Cryptosporidium parvum oocysts in test water (in this case natural lake water) in liquid solution. Candidate disinfectants that achieved substantive inactivation of oocysts in lake water (> 2-log inactivation in 8 hours) were evaluated with respect to published literature on their safety and with respect to their regulatory feasibility. In addition, they were adapted to granular solid disinfectant formulations and tabletized. Solid tablet disinfectants were evaluated with respect to their ability to achieve rapid inactivation (>= 3-log inactivation in < 4 hours) in simulated test waters comparable to those used in published EPA test procedures (i.e., Type I and Type II challenge waters). Candidate disinfectants that demonstrated rapid inactivation of Cryptosporidium parvum oocysts in test waters and that demonstrated potential regulatory feasibility were recommended for further testing and potential use.

Preparation of Clostridium sporogenes spores

[0058] Clostridium sporogenes spores were obtained from Mesa Labs (Lakewood,

CO). Stock spore suspension, at an initial concentration of 10⁷/ mL (stored in ethanol), was kept at 4°C until use. Before use, spores were centrifuged at 10,000xG for 10 minutes and resuspended in phosphate buffered saline (PBS) twice to remove ethanol. The resuspended spores were spiked into surface water from a nearby creek (Morgan Creek, Chapel Hill NC) at an approximate concentration of 5xl0³ spores/mL. The spiked surface water was aliquoted into 2-mL screw-capped vials (Sarstedt, Germany). Screw-capped vials were used as disinfection batch reactors for high-throughput testing because their small size enabled rapid and efficient screening while their airtight sealed screw caps enabled headspace and gas exchange to be minimized.

Preparation of Cryptosporidium parvum oocysts

[0059] Cryptosporidium parvum oocysts, Iowa strain (10⁸/ mL), were obtained from

University of Arizona parasitology lab (Tucson, AZ) and Waterborne, Inc. (New Orleans. LA). The production of the Iowa strain isolates consisted of propagation in neonatal calves approximately 1-2 a month. The oocysts were purified using discontinuous sucrose and cesium chloride density gradient centrifugation. The oocysts were stored at 4°C in antibiotic solution. Oocysts were centrifuged for 10 minutes at 10,000XG and resuspended in PBS 3x prior to use to eliminate the antibiotic solution. The resuspended oocysts were used within one week of centrifugation and all oocysts were used within three months of receipt. Oocysts were spiked into Morgan creek water or Type I or Type II challenge water at concentrations ranging from 10³-10⁵/ mL.

Challenge Water Matrix

[0060] Both natural and artificial challenge waters were used. Natural surface water was drawn from Morgan Creek, an urban stream in the town of Chapel Hill, Orange County, NC, prior to each experiment. Water was drawn from the surface at an access point on Culbreth Rd. in sterile 4-liter containers and immediately stored at 4°C. Type I and Type II test waters were prepared using stagnated UNC tap water as described by EPA (EPA, 1987, Guide standard and protocol for testing microbiological water purifiers). Clostridium perfringens spores or Cryptosporidium parvum oocysts were added to natural or artificial challenge waters at the required concentrations for each inactivation trial. Typically spores were added at concentrations of 10³-10⁴ CFU/mL and oocysts were added at concentrations of 10³- 10⁵ oocysts/mL.

Preparation of Candidate Disinfectants

Liquid (dissolved) disinfectants

[0061] A 1000X stock solution of NaDCC was prepared fresh daily by dissolving 976 mg of anhydrous NaDCC into 100 mL of deionized water. A 100X stock solution was prepared by diluting this 1:10 in deionized water; 20 pL of this lOOx solution was added to each 2-mL reactor to produce a final concentration of 9.76 mg/L NaDCC. At this dose, the estimated HOC1 concentration was 2.3 mg/L. According to the EPA, the maximum residual disinfectant level goal for free chlorine is 4 mg/L. The dose chosen was representative of the free chlorine dose typically used in household drinking water treatment and below the limit of 5 mg/L (McLaughlin et. al, 2009). Twenty pL of a lOOx metal ion solution, prepared the day before, containing 2200 mg/L zinc sulfate heptahydrate, 15.74 mg/L silver nitrate, and 390 mg/L copper sulfate pentahydrate were added to respective reactors to give final Zn⁺, Ag⁺, and Cu²⁺ ion concentrations of 5, 0.1, and 1 mg/L, respectively. All stock solutions were filter-sterilized using 0.22 pm syringe filters. Liquid disinfectant formulations (Table 1) were added to challenge waters at the beginning of each disinfection trial.

Table 1: Liquid Disinfectants.

Solid (Tablet) disinfectants

[0062] Granualar solid disinfectants were prepared (Table 2). Adipic acid was used as a compressible source of dry acid, and sodium carbonate and bicarbonate were added to produce an effervescent effect to aid in tablet dissolution. The ratio of adipic acid to carbonate/bicarbonate was selected to achieve pH < 6.5. Anhydrous NaDCC and/or copper, silver, and zinc in either salt or powdered form were also added to granular solid disinfectants as active ingredients. Sodium chlorite was also included in selected formulations. Granular solid disinfectants were compressed in a manual Carver model 3351 laboratory press using a 13 -mm tool and die set under 1 ton of pressure for approximately 1 minute to produce 13 -mm tablets for disinfection trials. A VICE hand-held tablet press was also used for some smaller batches of tablets with similar pressure and compression times. Tablets demonstrated adequate compressibility and dissolved within 30 minutes.

Table 2: Selected Disinfectant Tablet Formulations (mass of each constituent per batch of

25 tablets for disinfecting 1-L of water).

Neutralization

[0063] In order to conduct accurate disinfection trials, it is necessary to neutralize disinfectants at the time that samples are collected in order to separate the effects of disinfection during a defined contact time from any ongoing effects of residual disinfectants that might otherwise remain after that contact time. In order to develop a universal neutralizer for disinfectants used in this study, experiments were conducted to optimize concentrations of neutralizing substances used. For the neutralization of NaDCC, sodium thiosulfate (STS) was used. A neutralization experiment was performed with initial NaDCC concentration of 97.6 mg/L. Concentrations of Ox, lx (236 mg/L), 3x, lOx, 30x, and lOOx sodium thiosulfate were added to this concentration of NaDCC in 10 mL of distilled water. Hypochlorous acid (HOC1) concentration was tested using a color indicator test strip. Complete neutralization was observed at thiosulfate concentrations of 30x and lOOx. Similar trials were conducted with hydrogen peroxide and metals to evaluate suitable concentrations. A 30x molar excess of each neutralizing reagent was selected based on the results of these trials, and was found to stop the inactivation by E. coli by added disinfectants of interest. Final neutralizer solution consisted of 2,500 mg/L sodium thiosulfate (used to neutralize chlorine), 2,500 mg/L EDTA (used to chelate metal ions), 2,500 mg/L sodium pyruvate (used to neutralize H2O2, if present), added to phosphate buffered saline and filter-sterilized (0.22 um syringe filter). Aliquots (700 pL) were then pipetted into sterile 1.5 mL polypropylene tubes and stored at 4°C until use the next day. In some trials, higher concentrations of neutralizer were prepared and used in smaller aliquots to achieve the same final neutralizer concentration in neutralized samples.

Free Chlorine, pH, and metal ion Measurements

[0064] During experiments, the free chlorine concentration was monitored using the

DPD colorimetric method. The DPD method employed followed procedure 4500-Cl G (“DPD Colorimetric Method”) in the 18^th edition, Standard Methods for the Examination of Water and Wastewater. A standard calibration curve was prepared daily, and free chlorine concentrations determined based on this curve. A HACH Chemical reagent pillow containing N-diethyl-p-phenylenediamine (DPD) was added to 5 mL of PBS and 500 pL was aliquoted into 1.5 mL polypropylene tubes. Then, 500 pL of sample was added to the tube at corresponding time points and vortexed. 1 mL of the mixture was then added to a cuvette and absorbance was read at 515 nm using a Bio-Rad SmartSpec Plus (Hercules, CA). An adjustment was made to account for the dilution factor. Initial and final sample pH were determined using a benchtop pH meter. Interval time points (t=2, 4, 6 hours) were measured using pH strips for selected trials. Before and after selected experiments, aliquots of each sample point were reserved for metal ion analysis by inductively coupled plasma mass spectrometry (ICP-MS) to quantify metal ion concentrations over the duration of the experiment.

Disinfection Experiments

High-throughput screening with Clostridium Sporogenes spores

[0065] For high-throughput screening experiments involving Clostidium sporogenes spores, twenty pL of disinfectant solution (Table 1) was added to each 2-mL screw-cap vial containing C. sporogenes spores in Morgan Creek surface water. An initial 700-uL aliquot of the spiked reactors with disinfectant added was immediately collected and neutralized using an equivalent volume of neutralizer solution as described above to obtain the first sample at the t=0 time point. Additional aliquots were collected at the following time points: t =10, 20, 30, 40, 60, and 180 minutes. A separate reactor was sacrificed at each timepoint. During the experiment, reactors were covered with opaque paper or foil to block out light. The experiment was conducted at room temperature (20 to 25 °C). At each time point, 700 pL of sample for each disinfection condition was added to a 1.5 mL sterile Eppendorf tube containing 700 pL of neutralizer solution, specified below. The tubes were then vortexed for ~30 seconds to allow for proper neutralization. Serial dilutions were made in PBS or HEPES buffer and samples were stored at 4°C prior to analysis as described below.

[0066] Samples were enumerated for colonies by spot plate method on lx Reinforced

Clostridial Agar (RCA). Three spots (~10uL) were added to each plate per serial dilution (Fig. 1). Plates were placed in anaerobic jars with BD GasPak gas packs (BD, Franklin Lakes, NJ) and incubated at 35°C for 48 hours. After 48 hours, colonies were counted and colony forming units (CFU/ml) were calculated. Logio reduction values (LRV) were determined for each time point by subtracting the logio concentration at time t from the initial logio concentration at time t=0. Inactivation of Cryptosporidium parvum oocysts

Dissolved (liquid) disinfectants

[0067] At the start of each experiment, lmL of C. parvum oocysts suspended in PBS

(10⁷/mL) were spiked into the test water. Spiked test water (Morgan Creek surface water + oocysts) was distributed into 2 mL screw-capped vials. Screw-capped vials were used as disinfection batch reactors to minimize headspace and gas exchange, and thereby decrease potential of volatilization of disinfectant (observed in earlier trials using open systems). A separate vial was used for each time point of each condition (vials were sacrificed upon sampling) to further minimize volatilization from repeated opening and closing during sampling. Candidate disinfectants were added to vials at desired initial concentrations, and vials were mixed, as shown in Fig. 2. Negative controls (buffer) and positive controls (2 mL of test water with oocysts, no disinfectants) were also included.

[0068] An initial t=0 baseline sample of 700 pL was taken for each condition.

Samples of negative controls containing buffer and neutralizer, but no oocysts were also made. Small 1.5 mL polypropylene Eppendorf tubes were prepped with 700 pL of neutralizer mixture to cease disinfection at the time of sampling. Samples were collected and neutralized at t=0, 2,4,6, and 8 hours following disinfectant addition. At each time point, a 700-pL sample aliquot was pipetted from each 2-mL reactor into 1.5 mL polypropylene tubes containing neutralizer, and these neutralization tubes were vortexed for approximately 30 seconds to ensure mixing. During the experiment, reactor vials were kept covered to limit exposure to light. Following neutralization, 10-fold serial dilutions of neutralized samples were made in buffer. Neutralized samples were stored at 4°C prior to analysis.

Tablet disinfectants

[0069] At the start of each experiment, lmL of C. parvum oocysts suspended in PBS

(10⁸/mL) were spiked into each 4-L container of test water. Spiked test water (Type I or II test water + oocysts) was distributed into 1-L screw-capped glass bottles. Screw-capped bottles were used as disinfection batch reactors to minimize headspace and gas exchange, and thereby decrease potential of volatilization of disinfectant. A separate bottle was not used for each time point due to the prohibitive cost of using such a large quantity of oocysts. Candidate disinfectant tablets were added to 1-L reactors at the start of each experiment. Negative controls (test water with no oocysts and no disinfectants) and positive controls (test water with oocysts, no disinfectants) were also included. [0070] An initial t=0 baseline sample of 700 pL was taken for each condition. Small

1.5-mL polypropylene Eppendorf tubes were prepped with 700 pL of neutralizer mixture to cease disinfection at the time of sampling. Samples were collected and neutralized at t=0, 2, and 4 hours following disinfectant tablet addition. At each timepoint, a 700-pL sample aliquot was pipetted from each 1-L reactor into 1.5 mL polypropylene tubes containing neutralizer, and these neutralization tubes were vortexed for approximately 30 seconds to ensure mixing. In addition, at 2-h and 4-h time points, concentrated samples were prepared by collecting 50-mL to 250-mL aliquots from each reactor, neutralizing with 5-25 mL of 300X strength neutralizer (e.g. 10 times the strength used to neutralize the 700-pL aliquots), and centrifuging at 3000xG for 5 minutes to concentrate to a final volume of 1 mL. Following neutralization and/or concentration, 10-fold serial dilutions of neutralized samples were made in PBS or HEPES buffer. Neutralized samples were stored at 4°C prior to analysis.

Enumeration of Cryptosporidium Parvum

HCT-8 Cell Production and Maintenance

[0071] Cryptosporidium parvum is known to infect a range of continuous cell lines.

However, Human ileocecal colorectal adenocarcinoma (HCT-8) cells were used in this assay system as they are easily maintained and are sensitive to infection by C. parvum oocysts (Slikfo et ah, Applied and Environmental Microbiology, vol. 63, no. 9, Sept. 1997, pp. 3669- 3675). Cell stock, which contained approximately 1 mL of HCT-8 cells (stored at their 13^th passage) suspended in RPMI 1640 medium (GIBCO, Waltham, MA) was obtained from UNC Lineberger Tissue Culture Facility. Cell inocula were stored at -120°C and transported to lab in dry ice. Stock cell suspension was immediately thawed in 37°C water bath for approximately two minutes. The cell suspension was then added to 50 mL of HCT-8 maintenance medium (described elsewhere) in a T-75 corning flask. Flasks were incubated at 37°C for two days (+/- two days) before further cell passage to new cultures.

[0072] HCT-8 cells were maintained by bi-weekly serial passage in RPMI 1640 medium (Gibco), supplemented with heat-inactivated Fetal Bovine Serum (10% final concentration, Gibco), sodium pyruvate (ImM, Gibco), 100X antibiotics gentamycin (5 g/L) and kanamycin (25g/L), and 100X antifungal nystatin (2.5g/L) ( Simmons & Sobsey). Production was in T-75 and T-150 sterile tissue culture flasks (Coming, Corning, NY) and cells for assay were seeded onto Lab- Tek®II 8-well chamber slides (Thermo Fisher Scientific, Waltham, MA). For passage, media were first decanted from flasks and cells were washed once with phosphate buffer saline (PBS) in order to remove residual media. Next, lx EDTA-trypsin (Sigma, St. Louis, MI), was added to flasks. Flasks were then incubated at 37°C for 10 minutes. Flasks were further agitated by physically tapping and shaking to loosen cells. Next, trypsinized cells were aseptically decanted from flasks into sterile 50-mL polypropylene, conical bottom, screw-capped tubes (Corning Falcon, Tewksbury, MA) containing maintenance medium to neutralize the activity of the trypsin and EDTA. Trypsin/cell suspension was centrifuged for 8-10 minutes at 2,000XG. Supernatant was discarded and the cell pellet was resuspended in approximately 10 mL of maintenance medium. Cells were enumerated via hemocytometer and split into flasks or into 8-well chamber slides at a concentration of approximately 10⁶ cells/ mL.

[0073] For production of chamber slides and multiwell plates, cells were added to warmed media in a sterile beaker (media volume was calculated based on the number of chamber slides/wells to be prepared). The mixture was stirred on stir plate for approximately 10 minutes before dispensing into 8-well chamber slides or 24-well plates. Next, 400 pL of cell suspension were seeded into each chamber or well and allowed to settle under the hood for 15 minutes before being incubated at 37°C under a 5% CO2 atmosphere. Cells for assay were observed daily until 85% to 90% confluency was reached (typically within 72 +/- 24 hours).

HCT-8 Infectivity Assay -Immunofluorescent Microscopy

[0074] Samples of C. parvum oocysts were inoculated into HCT-8 cells for the immunofluorescent focus infectivity assay when adequate confluency of cells in chamber slides was reached. Briefly, media were aspirated without disrupting the cell layer and 100 pL of neutralized sample (from disinfection experiments) was inoculated onto the cell monolayer. Each disinfection condition/ dilution was plated in triplicate. Cells were incubated for two hours at 37°C, 5% CO2 to allow for C. parvum excy station and initial infection. Then, 500 pL of HCT-8 maintenance medium was added to each well and slides were incubated for an additional 48 to 72 hours (under the same conditions) to allow C. parvum living stages to continue to infect and develop within cell monolayers. The cell layers were then stained with an immunofluorescent-labeled monoclonal antibody (fluorescein isothiocyante- labeled C3C3 antibody; FITC C3C3 [an antibody that binds specifically to C. parvum living stages]) using methods similar to those previously described (Slikfo et al., 1997), with minor modifications. After incubation, the culture medium was aspirated and cell monolayers were fixed with absolute methanol (125 pL per well) for approximately thirty minutes at room temperature. The methanol was aspirated and the cell layers were washed with lx phosphate-buffered saline (PBS) three times. Next, blocking buffer (0.25 mL per well) consisting of PBS and 1 mg/L bovine serum albumin (BSA) was added to each chamber for approximately thirty minutes at room temperature. Blocking buffer was aspirated and monolayers were stained with 75 pL of purified FITC-C3C3, diluted in PBS-BSA at 1:300. Wells were covered in foil and placed on a shaking platform for 90 minutes at room temperature. Antibody solution was aspirated, and slide cell layers were washed with PBS three times. Chambers from slides were removed, 65 pL PVA/DABCO mounting medium was added to each slide and 22x50 mm coverslips were mounted. Transparent nail polish was added to seal edges of the cover slip.

[0075] Slides were observed under an oil emersion lens at a total magnification of

400x using a Leitz Orthoplan 2 fluorescent microscope. Slides were scanned in a systematic fashion and infectious foci were enumerated either by direct count or by a most probable number (MPN) method. In the direct count method, infectious foci were counted and recorded. In the MPN method, each viewing field was scored as a positive or negative for apple-green fluorescing ovoid or spherical objects approximately 3 to 6 pm in diameter. A most probable number (MPN) of infectious units (UI) per mL calculation was conducted (Jarvis B., et ah, (2010) Reconsideration of the derivation of Most Probable Numbers, their standard deviations, confidence bounds and rarity values. Journal of Applied Microbiology doi: 10.1111/j.1365-2672.2010.04792.x) . The summary of the HCT-8 infectivity assay for C. parvum can be seen in Fig. 3.

MPN (UI/mL)=

[0076] Infectious foci were counted using a fluorescent microscope. Images from disinfection that indicated infectious foci are shown in Fig. 4.

HCT-8 Infectivity Assay -Integrated Cell culture-PCR

[0077] Samples of C. parvum oocysts were inoculated into HCT-8 cells for the integrated cell culture-PCR infectivity assay when confluency of cells in 24-well plates was reached. Briefly, media were aspirated without disrupting the cell layer and 100 pL of neutralized sample (from disinfection experiments) was inoculated onto the cell monolayer. Each disinfection condition/ dilution was plated in triplicate. Cells were incubated for two hours at 37°C, 5% CO2 to allow for C. parvum excystation and initial infection. Then, 500 pL of HCT-8 maintenance medium was added to each well and slides were incubated for an additional 48 to 72 hours (under the same conditions) to allow C. parvum living stages to continue to infect and develop within cell monolayers. The cell layers were then aspirated to remove maintenance medium and the cell layers were washed with lx phosphate-buffered saline (PBS) three times. Next, cells were harvested by manual pipetting and centrifuged for 5 minutes at 10,000xG. The supernatant was discarded and the pellet containing HCT-8 cells and C. parvum living stages (if infection occurred) was retained. Nucleic acid was then extracted from each cell pellet using a Qiagen (Hilden, Germany) QIAamp DNA mini-kit according to the manufacturer’s instructions and amplified by PCR using a Bio-Rad CFX96 Touch thermocycler. Primers were used as described by Di Giovanni et al. (1999) Appl. Environ. Microbiol ., 65(8), 3427-3432. Thermocycler conditions were: 95°C for 10 min, 95°C for 30 sec, 60°C for 60 sec, 72°C for 60 sec, repeat for 45 cycles, 72°C for 7 min, and hold at 4°C. Concentrations of infectious C. par um oocysts in the original sample were determined based on the number of cell culture wells in which C. parvum DNA was detected after washing (e.g. indicating that a positive infection had occurred) using an MPN approach as described above. Positive and negative controls were also included, and a standard curve was run to confirm that the method amplified C. parvum DNA from infected monolayers but did not amplify trace residual DNA from noninfectious oocysts or negative controls.

Data Analysis

[0078] Data were analyzed using Microsoft Excel (Microsoft Corporation, Redmond,

WA) and STATA SE (StataCorp, College Station, TX). Summary statistics were calculated and log reduction values were computed for inactivation trials.

Results

High-throughput Screening

[0079] Disinfection experiments were conducted with C. sporogenes spores as an indicator for C. parvum infectivity in Morgan Creek water buffered to pH 5.0 (for maximum inactivation in initial screening). Reactors included either minimal headspace or 50% headspace. Rapid inactivation of C. sporogenes spores was observed in surface water, as shown in Table 3. An average 1.7-log reduction was achieved with NaDCC along and a >3.8 log reduction with candidate disinfectant 3-L (NaDCC + metal ions) after 10 minutes. There were no detectable C. sporogenes colonies remaining after 10 minutes of contact with candidate disinfectant 3-L, but colonies were detected after 10 minutes incubation with NaDCC. After 30 minutes, no C. sporogenes colonies were detected for either disinfectant. Reported logio reduction values for candidate disinfectant 3-L after 10 minutes and for NaDCC alone after 30 minutes are based on lower detection limits, and could therefore be greater than the numeric values reported.

Table 3: Logio reduction values of C. sporogenes spores after incubation for 10 minutes in the presence of NaDCC and candidate disinfectant 3-L (NaDCC +Metals) (n=3).

[0080] The effect of head-space was also pronounced. C. sporogenes was not detected after ten minutes of contact time with candidate disinfectant 3-L, corresponding to a >3.78 logio reduction, in reactors containing no headspace. No substantive inactivation was observed after 20 minutes of contact time in reactors with headspace, and complete inactivation in headspace reactors did not occur after 60 minutes (Table 4).

Table 4: Log Reduction values of C. sporogenes spores for candidate disinfectant 3-L (NaDCC + M) with no headspace and 50% headspace (n=3)

Liquid disinfectants

[0081] Liquid disinfectant trials were conducted in stagnated tap water (typical conditions) and Morgan Creek water (high organic conditions) to determine log reduction in infectivity of C. parvum oocysts incubated for eight hours in the presence of 1) NaDCC; 2) Metal ions; and 3) Candidate disinfectant 3-L consisting of NaDCC and metal ions. Preliminary trials using a single replicate in stagnated tap water buffered to pH 5.0 showed 1.3-log inactivation by NaDCC alone in 8 hours and >2 log inactivation with candidate disinfectant 3-L after 8 hours (Table 5).

Table 5: Preliminary log reduction values calculated using MPN for C. parvum oocysts by NaDCC (1-L), candidate disinfectant (3-L, NaDCC + metals) and other formulations in stagnated tap water after eight hours of disinfection. (n =1)

[0082] However, follow-up experiments in Morgan Creek water (pH 5) using triplicate reactors showed 1.2 log inactivation of C. parvum using NaDCC alone, and 1.4-log inactivation of C. parvum oocysts in creek water using candidate disinfectant 3-L (Table 6). The metals-only condition (2-L) had an average logio reduction of 1.2.

Table 6: Log reduction values (with standard deviation) calculated using MPN for C. parvum oocysts by NaDCC (1-L), metal ions (2-L) and candidate disinfectant (3-L, NaDCC + metals) in Morgan Creek water after eight hours of disinfection (n =3)

Tablet Disinfectants

[0083] In order to evaluate the potential of the candidate disinfectants to inactivate C parvum in test waters similar to those recommended in the EPA Guide Standard protocols, tabletized disinfectants were used to inactivate C. parvum oocysts in 1-L samples of Type I (low-turbidity and pH 7, Table 7) and Type II (high-turbidity and pH 9, Table 8) test water. Extensive inactivation of Cryptosporidium parvum oocysts was observed in Type I test water (more characteristic of drinking water conditions). Somewhat less extensive inactivation was observed in Type II test water (more characteristic of worst-case conditions).

Table 7: Log reduction values (with standard deviation) calculated using MPN for C. parvum oocysts by NaDCC and candidate disinfectants in Type-I challenge water (1-L reactors) after two hours of disinfection (n =3)

Table 8: Log reduction values (with standard deviation) calculated using MPN for C. parvum oocysts by NaDCC and candidate disinfectants in Type-II challenge water (1-L reactors) after two hours of disinfection (n =3)

[0084] The foregoing is illustrative of the present invention, and is not to be construed as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein. All publications, patent applications, patents, patent publications, and other references cited herein are incorporated by reference in their entireties for the teachings relevant to the sentence and/or paragraph in which the reference is presented.

Claims

THAT WHICH IS CLAIMED IS

1. A composition comprising: a halogen-containing compound; and a second active agent, wherein the second active agent is selected from the group consisting of a metal, a chlorite, a chlorate, and combinations thereof.

2. The composition of claim 1, wherein the halogen-containing compound is a dichloroisocyanurate (e.g., sodium dichloroisocyanurate (NaDCC)) and/or a trichloroisocyanurate (e.g., sodium trichloroisocyanurate).

3. The composition of claim 1, wherein the halogen-containing compound is a bromine-containing compound (e.g., sodium dibromoisocyanurate and/or another dibromoisocyanurate) and/or an iodine-containing compound, optionally wherein the halogen-containing compound is a dibromoisocyanurate (e.g., sodium dibromoisocyanurate) and/or a tribromoisocyanurate (e.g., sodium tribromoisocyanurate).

4. The composition of any preceding claim, wherein the halogen-containing compound is NaDCC and the composition comprises at least one additional halogen- containing compound.

5. The composition of any preceding claim, wherein the halogen-containing compound is a compound that provides a source of free chlorine, a source of combined chlorine, a source of monochloramine, a source of dichloramine, a source of other chloramines (e.g., chloramine-T and/or chloramine-B), a source of chlorine dioxide, a source of bromine, and/or a source of iodine.

6. The composition of any preceding claim, wherein the halogen-containing compound in present in the composition in a biocidal amount.

7. The composition of any preceding claim, wherein the second active agent comprises a metal (e.g., copper, silver, and/or zinc), optionally wherein the metal is present in the composition in the form of a metal salt and/or metal powder.

8. The composition of any preceding claim, wherein the metal is present in the composition in an amount of about 0.0001 mg/L to about 100 mg/L and/or wherein the metal is present in the composition in an amount of about 0.001% to about 10% by weight of the composition.

9. The composition of any preceding claim, wherein the metal and the halogen- containing compound (e.g., NaDCC) are present in the composition in a ratio, by weight, of about 1:10000 to about 10:1 (metal : the halogen-containing compound).

10. The composition of any preceding claim, wherein the metal comprises copper.

11. The composition of claim 10, wherein copper is present in an amount of about 0.01, 0.05, 0.1, 0.5, 1, or 2 mg/L or more, and/or wherein copper is present in the composition in an amount of about 0.0001% to about 10% by weight of the composition, and/or wherein copper and the halogen-containing compound (e.g., NaDCC) are present in the composition in a ratio, by weight, of about 1:10000 to about 10:1 (copper : the halogen- containing compound ).

12. The composition of any preceding claim, wherein the metal comprises silver.

13. The composition of claim 12, wherein silver is present in an amount of about 0.00001 mg/L to about 2 mg/L, and/or wherein silver is present in the composition in an amount of about 0.0001% to about 10% by weight of the composition, and/or wherein silver and the halogen-containing compound (e.g., NaDCC) are present in the composition in a ratio, by weight, of about 1:10000 to about 10:1 (silver : the halogen- containing compound).

14. The composition of any preceding claim, wherein the metal comprises zinc.

15. The composition of claim 14, wherein zinc is present in an amount of about 0.01 mg/L to about 1000 mg/L, and/or wherein zinc is present in the composition in an amount of about 0.001% to about 10% by weight of the composition, and/or wherein zinc and the halogen-containing compound (e.g., NaDCC) are present in the composition in a ratio, by weight, of about 1:1000 to about 100:1 (zinc : the halogen- containing compound).

16. The composition of any preceding claim, wherein the second active agent is a chlorite or a chlorate, optionally wherein the chlorite is sodium chlorite and/or potassium chlorite, and/or optionally wherein the chlorate is sodium chlorate and/or potassium chlorate.

17. The composition of claim 16, wherein the chlorite and/or chlorate is present in an amount of about 0.001 mg/L to about 1 mg/L, and/or wherein the chlorite and/or chlorate is present in the composition in an amount of about 0.001% to about 10% by weight of the composition, and/or wherein the chlorite and/or chlorate and the halogen-containing compound (e.g., NaDCC) are present in the composition in a ratio, by weight, of about 1 : 1000 to about 1 : 1 (chi orite/chl orate : the halogen-containing compound).

18. The composition of any preceding claim, wherein the halogen-containing compound (e.g., NaDCC) is present in the composition in an amount of about 0.001 or 0.01 mg/L to about 10 or 100 mg/L and/or wherein the halogen-containing compound is present in the composition in an amount of about 0.001% to about 10% or 25% by weight of the composition.

19. The composition of any preceding claim, wherein the halogen-containing compound (e.g., NaDCC) is present in an anhydrous form, dihydrate form, or other hydrated form.

20. The composition of any preceding claim, further comprising a carbonate, a bicarbonate, and/or an acid (e.g. a solid organic acid or another dry acid, optionally sodium bisulfate), optionally wherein the acid is adipic acid.

21. The composition of any preceding claim, further comprising a buffering agent (e.g., an acid, base, etc.), optionally wherein the buffering agent is present in the composition in an amount to provide a pH of about 2 to about 7 and/or to provide a buffer strength of about 0.1 mM to about 10 M.

22. The composition of any preceding claim, further comprising a coagulant, optionally wherein the coagulant is a chitosan (e.g., chitosan acetate).

23. The composition of claim 22, wherein the coagulant is present in an amount of about 0.01 mg/L to about 100 mg/L, and/or wherein the coagulant is present in the composition in an amount of about 0.001% to about 10% by weight of the composition, and/or wherein the coagulant and the halogen-containing compound (e.g., NaDCC) are present in the composition in a ratio, by weight, of about 1 : 1000 to about 100: 1 (coagulant: the halogen-containing compound).

24. The composition of any preceding claim, further comprising one or more additive(s), optionally wherein the one or more additive(s) comprise a surfactant, a source of peroxide (e.g., a percarbonate), a source of peracetic acid (e.g., hydrogen peroxide or a source of peroxide such as a percarbonate, optionally sodium percarbonate plus a bleach activator, optionally tetraacetylethylenediamine (TAED)), and/or a source performic acid (e.g. hydrogen peroxide or a source of peroxide such as a percarbonate, optionally sodium percarbonate plus a source of formic acid, optionally formic acid or sodium performate), optionally in the presence of an additional catalyst such as an iron salt.

25. The composition of any preceding claim, wherein the composition is in the form of one or more tablet(s), optionally wherein the one or more tablet(s) dissolve in about 1 liter, 5 liters, 10 liters, and/or 20 liters of a liquid (e.g., water) and/or dissolve in a 1 liter, 5 liter, 10 liter, and/or 20 liter container comprising a liquid (e.g., water) in an amount of about 1 liter, 5 liters, 10 liters, and/or 20 liters, respectively, and/or in an amount that provides less than about 5% headspace in the container, optionally at room temperature (e.g., about 20°C to about 22°C) and/or in about 1, 2, 5, 10, or 15 minute(s) to about 20, 25, 30, 35, 40, 45, 50, 55 or 60 minutes after contact of the one or more tablet(s) and the liquid.

26. The composition of any one of claims 1-24, wherein the composition is a liquid.

27. The composition of any one of claims 1-24, wherein the composition is in the form of a powder and/or granules.

28. The composition of any one of claims 1-24, wherein the composition is in the form of a sachet and/or dispersible capsule or pod.

29. The composition of any preceding claim, wherein the composition has and/or in 1 L of water has a pH of about 6.5 or less.

30. The composition of any preceding claim, wherein the composition has and/or provides a biocidal concentration of chlorine, and/or wherein chlorine is present as hypochlorous acid, hypochlorite, and/or combined chlorine (e.g., monochloramine, dichloramine, etc.).

31. The composition of any preceding claim, wherein the composition has and/or provides a biocidal concentration of bromine, and/or wherein bromine is present as hypobromous acid, hypobromite, and/or combined bromine (e.g., monobromamine, dibromamine, etc.).

32. The composition of any preceding claim, wherein the composition has and/or provides a biocidal concentration of iodine, and/or wherein iodine is present as hypoiodous acid and/or hypoiodite.

33. The composition of any preceding claim, wherein the composition has and/or provides a biocidal concentration of chlorine dioxide.

34. The composition of any preceding claim, wherein the composition has and/or provides a biocidal concentration of two or more oxidants, optionally wherein at least one of the two or more oxidants is selected from the group consisting of a peroxide, a peracetic acid, a performic acid, a reactive halogen species such as free chlorine (e.g., hypochlorous acid, hypochlorite, etc.), hypobromous acid, hypobromite, hypoiodous acid, hypoiodite, a chloramine, a bromamine, an iodamine, a reactive oxygen species (e.g., superoxide radical anion and/or hydroxyl radical), chlorine dioxide.

35. The composition of any preceding claim, wherein the composition provides a release of a halogen containing compound, halogen, and/or biocidal compound for a period of time (e.g., for about 1, 10, 30, or 45 minutes or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15,

20, or 24 hours or more), optionally wherein the composition provides a release (e.g., continuous release) of chlorine, chlorine dioxide, HOC1, a chloramine, bromine, a bromamine, iodine, and/or an iodamine for a period of time.

36. A method of inactivating and/or killing a microorganism, the method comprising: exposing a microorganism to a composition of any one of claims 1 to 35, thereby inactivating and/or killing the microorganism.

37. A method of inactivating and/or killing a microorganism, the method comprising: exposing a microorganism to a composition comprising a halogen-containing compound (e.g., NaDCC) to form a mixture, wherein the mixture has a pH of less than about 6.5, thereby inactivating and/or killing the microorganism.

38. The method of claim 36 or 37, wherein the halogen-containing compound (e.g., NaDCC) is present in the composition in an amount of about 0.001 mg/L to about 100 mg/L and/or wherein the halogen-containing compound (e.g., NaDCC) is present in the composition in an amount between about 0.001% to about 10% or 25% by weight of the composition.

39. The method of any one of claims 36-38, wherein the composition is a composition of any one of claims 1 to 35.

40. The method of any one of claims 36-39, wherein the microorganism is present in water and the method further comprises disinfecting the water.

41. The method of any one of claims 36-40, wherein the microorganism is present on a surface of an article of manufacture or composition and the method further comprises disinfecting the surface of the article of manufacture or composition.

42. The method of any one of claims 36-41, wherein the microorganism is present in a food and/or beverage and the method further comprises disinfecting the food and/or beverage.

43. The method of any one of claims 36-42, wherein the microorganism is a Cryptosporidium species, a Clostridium species, and/or a coronavirus.

44. The method of any one of claims 36-43, wherein the method inactivates and/or kills Cryptosporidium parvum oocysts.

45. The method of any one of claims 36-44, wherein the exposing step is carried out in a container comprising the composition and microorganism and the container has less than about 50%, 40%, 30%, 20%, 10%, 5% or 1% headspace.

46. The method of any one of claims 36-45, wherein the composition is in the form of a solid (e.g., tablet, granules, etc.), and the solid dissolves in about 1 minute to about 5, 10, 15, 20, 30, 45, or 60 minutes, optionally at a temperature in a range of about 15°C to about 25°C or at about room temperature, optionally wherein the solid is one or more tablet(s) that dissolve in about 1 liter, 5 liters, 10 liters, and/or 20 liters of a liquid (e.g., water) and/or dissolve in a 1 liter, 5 liter, 10 liter, and/or 20 liter container comprising a liquid (e.g., water) in an amount of about 1 liter, 5 liters, 10 liters, and/or 20 liters, respectively, and/or in an amount that provides less than about 5% headspace in the container.

47. The method of any one of claims 36-46, wherein the method provides water suitable for drinking and/or animal (e.g., human, livestock, etc.) consumption.