Neutralizing compositions and related methods

Abstract

The present disclosure relates to compositions and methods for the neutralization of antimicrobial compounds. The neutralizing compositions include at least one compound capable of neutralizing at least one antimicrobial compound and at least one compound suitable for aiding in the recovery of one or more microorganisms.

Description

NEUTRALIZING COMPOSITIONS AND RELATED METHODSFIELD

[0001] The present disclosure relates to compositions and methods for the neutralization of antimicrobial compounds.BACKGROUND

[0002] Various industries require sampling of surfaces to determine the presence or absence, and in some cases quantification, of certain microorganisms. Generally, these industries will clean or sanitize surfaces with harsh antimicrobial agents prior to sampling of the surface. These tests can show whether cleaning or sanitation procedures are sufficient. Residual antimicrobial agents can, however, interfere with the culturing or detection of microorganisms even when the microorganisms are not killed by the antimicrobial agents. The result is that false negatives may be observed wherein a live microorganism of interest is present on the surface, but it is not detected in a culture because of the antimicrobial agents interfere with the culturing or detection. Such false negative can result in the mistaken belief that surfaces are sufficiently free of microorganisms when, in fact, the surfaces have microorganisms that are left behind and may transfer to another surface or host.BRIEF SUMMARY OF THE DISCLOSURE

[0003] The present disclosure relates to compositions and methods for the neutralization of antimicrobial compounds. The compositions provided herein may neutralize an antimicrobial compound in the presence of microorganisms so that one or more microorganisms not killed by the antimicrobial agent can be recovered and cultured. The compositions provided herein may neutralize any residual antimicrobial agents in a test sample while not adversely impacting downstream detection assays.

[0004] According to one aspect, a neutralizing composition is provided that includes at least one compound capable of neutralizing at least one antimicrobial compound and at least one compound suitable for aiding in the recovery of one or more microorganisms. According to one embodiment, the at least one compound capable of neutralizing at least one antimicrobial compound includes a polyethylene sorbitol ester. According to one embodiment, the polyethylene sorbitol ester includes polyoxyethylene sorbitan monooleate. According to one embodiment, the at least one compound capable of neutralizing at least one antimicrobial compound includes lecithin. According to one embodiment, the at least one compound capable of neutralizing at least one antimicrobial compound includes casein peptone type 1 , meatpeptone, beef extract, or any combination thereof. According to one embodiment, the at least one compound capable of neutralizing at least one antimicrobial compound includes L-histidine, sodium thiosulfate, or a combination thereof. According to one embodiment, the at least one compound capable of neutralizing at least one antimicrobial compound includes sodium bisulfite. According to one embodiment, the at least one compound capable of neutralizing at least one antimicrobial compound includes sodium carbonate. According to one embodiment, the at least one compound suitable for aiding in the recovery of one or more microorganisms includes a metal halide. According to one embodiment, the at least one compound suitable for aiding in the recovery of one or more microorganisms includes magnesium chloride (MgCI). According to one embodiment, the neutralizing composition further includes at least one yeast extract. According to one embodiment, the neutralizing composition further includes sodium phosphate dibasic. According to one embodiment, the neutralizing composition further includes at least one antifoaming compound. According to one embodiment, the at least one antifoaming compound is antifoam powder. According to one embodiment, the neutralizing composition is shelf stable at ambient conditions for at least one week. According to one embodiment, the neutralizing composition has a pH of from about 6 to about 8.

[0005] According to another aspect, a neutralizing composition is provided that includes or consists essentially of: about 5.0 g / L to about 15 g / L of lecithin; about 10.0 g / L to about 20.0 g / L of polyoxyethylene sorbitan monooleate; about 0.1 g / L to about 0.5 g / L of at least one antifoaming compound; and about 0.1 g / L to about 0.5 g / L of magnesium chloride (MgCI). According to one embodiment, the neutralizing composition further includes one or more of peptone type 1 , meat peptone, beef extract, L-histidine, sodium thiosulfate, sodium bisulfite, sodium carbonate, at least one yeast extract, or sodium phosphate dibasic. According to one embodiment, the at least one antifoaming compound is antifoam powder. According to one embodiment, the neutralizing composition is shelf stable at ambient conditions for at least one week. According to one embodiment, the neutralizing composition has a pH of from about 6 to about 8.

[0006] According to one aspect, vials for downstream detection assays are provided. The vials provided herein may be particularly suitable for use with devices for rapid detection of microorganisms. The vials may also include a neutralizing composition as provided herein.

[0007] According to one aspect, a method of neutralizing at least one antimicrobial compound in a culture medium is provided and includes the step of introducing an effective amount neutralizing composition as provided herein to a culture media.

[0008] According to one aspect, a method of culturing for one or more microorganisms is provided. The method includes the steps of: sampling a surface to remove a sample of the one or more microorganisms from the surface to form a test sample; mixing the test sample with culture medium to form a test sample composition; and contacting the test sample composition with a neutralizing composition as provided herein so as to allow one or more microorganisms to multiply within the culture medium.BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 sets forth a table providing detection times when various levels of sodium bisulfite, sodium thiosulfate, sodium thioglycollate, lecithin, sodium chloride, magnesium chloride, and histidine are added to letheen broth.DETAILED DESCRIPTION

[0010] It is to be understood that the methods described in this disclosure are not limited to particular methods and experimental conditions disclosed herein, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.Furthermore, the experiments described herein, unless otherwise indicated, use conventional techniques within the skill of the art.

[0011] Unless otherwise defined herein, scientific and technical terms used herein have the meanings that are commonly understood by those of ordinary skill in the art. In the event of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. Unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. The use of “or” means “and / or” unless stated otherwise. The use of the term “including”, as well as other forms, such as “includes” and “included”, is not limiting.

[0012] Unless otherwise defined, all terms of art, notations, and other scientific terms or terminology used herein are intended to have the meanings commonly understood by those of skill in the art to which the application pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art.Definitions

[0013] As used herein, the following terms have the meanings as ascribed to them below, unless specified otherwise.

[0014] Unless specifically stated or obvious from context, as used herein, the term “or” is understood to be inclusive. Unless specifically stated or obvious from context, as used herein, the terms “a”, “an”, and “the” are understood to be singular or plural.

[0015] The term “comprise”, “comprises”, and “comprising” will be understood to imply the inclusion of a stated step or component or group of steps or components but not the exclusion of any other step or element or group of steps or elements. By “consisting of” is meant to include, and be limited to, whatever follows the phrase “consisting of.” Thus, the phrase “consisting of” indicates that the listed elements are required or mandatory, and that no other elements may be present. By “consisting essentially of” is meant including any elements listed after the phrase, and be limited to other elements that do not interfere with or contribute to the activity or action specified in the disclosure for the listed elements.

[0016] The terms “neutralize,” “deactivate,” “inactivate” and derivatives thereof may be used interchangeably and refer to the reduction or elimination of interference caused by antimicrobial compounds with regard to culturing or detection of microorganisms.

[0017] The terms “antimicrobial agent” and “antimicrobial compound” may be used interchangeably and refer to substances such as disinfectants for destroying or inhibiting the growth of microorganisms on a surface. Antimicrobial compounds as provided herein may include compounds commonly found in cosmetics, personal care products, household cleaners, nutraceuticals, cannabis formulations, and disinfectant sprays. Such antimicrobial compounds include, but are not limited to, quaternary ammonium compounds, organic acids, halogens, alcohols, diureides. aldehydes, parabens, sorbates, aldehydes, mercurials, biguanides, glucosides, acrylates, organo-mercuric compounds, isothiazolinones, imidazoles, and phenolics. Antimicrobial compounds as provided herein may include compounds commonly found in personal care formulations or cosmetics such as acrylates, coal tar, salicylic acid, sulfur, alloantoin, potassium sorbate, phenoxyethanol, decyl glucoside, and citric acid.

[0018] The term “microorganism” refers to a living organism that lives on a surface including, for example, bacteria, viruses, molds or fungi. Typical bacteria found on a surface include, but are not limited to, Enterobacteriaceae, Alicyclobacillus, Pseudomonas (e.g., Pseudomonas aeruginosa), Staphylococcus (e.g., Staphylococcus aureus), Enterococcus (e.g., Enterococcus hirae), E. coli, Salmonella (e.g., Salmonella enterica serovar Typhimurium), Listeria monocytogenes, or a mixture thereof.

[0019] The term “effective amount” refers to the sufficient amount of neutralizing composition to neutralize the activity of one or more antimicrobial compounds present, or suspected of being present, in a culture media. The term may also refer to the amount of a component present in the neutralizing composition sufficient to carried out the component’s intended purpose.

[0020] The terms “culture media” and “growth media” may be used interchangeably and refer to a composition used to aid the growth and cultivation of microorganisms. The culture media may include a liquid nutrient medium / broth (e.g., agar) that provides nutrients and water to microorganisms. Growth media may include thin film growth media, such as those available as PETRIFILM® from 3M Company of St Paul, Minn., USA and compatible with agar-based media.Neutralizing Compositions

[0021] Neutralizing compositions are provided that neutralize an antimicrobial compound in the presence of microorganisms so that one or more microorganisms not killed by the antimicrobial agent can be recovered and cultured. The neutralizing compositions provided herein may neutralize any residual antimicrobial agents in a test sample while not adversely impacting downstream detection assays.

[0022] The neutralizing compositions provided herein are shelf stable at ambient or room temperature conditions (e.g., about 15°C to about 29°C) for at least one week, two weeks, three weeks, four weeks, five weeks, six weeks, seven weeks, eight weeks, nine weeks ten weeks, eleven week, or even twelve weeks or more. The neutralizing compositions provided herein are shelf stable at elevated temperatures (e.g., about 30°C to about 40°C) for at least one week, two weeks, three weeks, four weeks, or even five weeks or more. The neutralizing compositions provided herein are shelf stable at reduced temperatures (e.g., about 2°C to about 10°C) for at least one week, two weeks, three weeks, four weeks, or even five weeks or more.

[0023] According to one embodiment, the neutralizing compositions provided herein include at least one compound suitable for neutralizing phenolic compounds. Any suitable one or more peptones can be used for neutralizing phenolic compounds. The one or more peptones may include one or more of soytone, tryptone, porcine peptone, casein peptone type 1 , meat peptone, or beef extract.

[0024] The at least one compound suitable for neutralizing phenolic compounds may be present in the neutralizing composition at an effective amount suitable for neutralizing phenolic compounds. According to one embodiment, the effective amount of at least one compoundsuitable for neutralizing phenolic compounds present in the neutralizing compositions provided herein, in g / L, may be 0.10 or more, 1 .0 or more, 2.25 or more, 5.0 or more, 7.5 or more, 10.0 or more, 15.0 or more, 20.0 or more, 25.0 or more, or even 30.0 or more. The effective amount of at least one compound suitable for neutralizing phenolic compounds present in the neutralizing compositions provided herein, in g / L, may be 30.0 or less, 25.0 or less, 20.0 or less, 15.0 or less, 10.0 or less, 7.5 or less, 5.0 or less, 2.25 or less, 1 .0 or less, or even 0.10 or less.

[0025] According to one embodiment, the neutralizing compositions provided herein include at least one primary yeast extract. The primary yeast extract is suitable for serving as a nitrogen source for microorganisms while providing clarity and brightness suitable for downstream assays. The at least one primary yeast extract may be obtained by the autolysis of a yeast strain grown on a molasses-based media such as ProCel® 351 PW available from Procelys.

[0026] The at least one primary yeast extract may be present in the neutralizing composition at an effective amount. According to one embodiment, the effective amount of primary yeast extract present in the neutralizing compositions provided herein, in g / L, may be 0.1 or more, 1 .0 or more, 1 .5 or more, 2.0 or more, 2.5 or more, or even 3.0 or more. The effective amount of primary yeast extract in the neutralizing compositions provided herein, in g / L, may be 3.0 or less, 2.5 or less, 2.0 or less, 1 .5 or less, 1 .0 or less, or even 0.1 or less.

[0027] According to one embodiment, the neutralizing compositions provided herein include at least one salt. An exemplary salt includes sodium chloride. The at least one salt may be present in the neutralizing composition at an effective amount. According to one embodiment, the effective amount of the at least one salt present in the neutralizing compositions provided herein, in g / L, may be 1 .0 or more, 2.5 or more, 5.0 or more, or even 7.5 or more. The effective amount of salt in the neutralizing compositions provided herein, in g / L, may be 7.5 or less, 5.0 or less, 2.5 or less, or even 1 .0 or less.

[0028] According to one embodiment, the neutralizing compositions provided herein include at least one compound suitable for neutralizing aldehyde compounds. According to one embodiment, the at least one compound suitable for neutralizing aldehyde compounds includes at least one essential amino acid. According to one embodiment, the at least one compound suitable for neutralizing aldehyde compounds includes L-histidine.

[0029] The at least one compound suitable for neutralizing aldehyde compounds may be present in the neutralizing composition at an effective amount. According to one embodiment, the effective amount of the at least one suitable for neutralizing aldehyde compounds present in the neutralizing compositions provided herein, in g / L, may be 0.10 or more, 0.25 or more, 0.50or more, 0.75 or more, 1 .00 or more, or even 1 .25 or more. According to one embodiment, the effective amount of the at least one suitable for neutralizing aldehyde compounds present in the neutralizing compositions provided herein, in g / L, may be 1 .25 or less, 1 .00 or less, 0.75 or less, 0.50 or less, 0.25 or less, or even 0.10 or less.

[0030] According to one embodiment, the neutralizing compositions provided herein include at least one compound suitable for neutralizing quaternary ammonium compounds. The at least one compound suitable for neutralizing quaternary ammonium compounds includes lecithin.

[0031] The at least one compound suitable for neutralizing quaternary ammonium compounds may be present in the neutralizing composition at an effective amount suitable. According to one embodiment, the effective amount of at least one compound suitable for neutralizing quaternary ammonium compounds present in the neutralizing compositions provided herein, in g / L, may be 0.10 or more, 1 .0 or more, 2.25 or more, 5.0 or more, 7.5 or more, 10.0 or more, or even 15.0 or more. The effective amount of at least one compound suitable for neutralizing quaternary ammonium compounds present in the neutralizing compositions provided herein, in g / L, may 15.0 or less, 10.0 or less, 7.5 or less, 5.0 or less, 2.25 or less, 1 .0 or less, or even 0.10 or less.

[0032] According to one embodiment, the neutralizing compositions provided herein include at least one compound suitable for neutralizing formaldehyde and glutaraldehyde compounds. The at least one compound suitable for neutralizing formaldehyde and glutaraldehyde compounds includes sodium bisulfite.

[0033] The at least one compound suitable for neutralizing formaldehyde and glutaraldehyde compounds may be present in the neutralizing composition at an effective amount. According to one embodiment, the effective amount of the at least one suitable for neutralizing formaldehyde and glutaraldehyde compounds present in the neutralizing compositions provided herein, in g / L, may be 0.01 or more, 0.05 or more, 0.10 or more, or even 0.25 or more. According to one embodiment, the effective amount of the at least one suitable for neutralizing formaldehyde and glutaraldehyde compounds present in the neutralizing compositions provided herein, in g / L, may be 0.25 or less, 0.10 or less, 0.05 or less, or even 0.01 or less.

[0034] According to one embodiment, the neutralizing compositions provided herein include at least one compound suitable for neutralizing phenols, hexachlorophene, formalin, and ethanol. The at least one compound suitable for neutralizing phenols, hexachlorophene, formalin, and ethanol includes at least one polyethylene sorbitol ester. According to a particularembodiment, the least one compound suitable for neutralizing phenols, hexachlorophene, formalin, and ethanol includes polyoxyethylene sorbitan monooleate.

[0035] The at least one compound suitable for neutralizing phenols, hexachlorophene, formalin, and ethanol may be present in the neutralizing composition at an effective amount suitable for neutralizing phenolic compounds. According to one embodiment, the effective amount of at least one compound suitable for neutralizing phenols, hexachlorophene, formalin, and ethanol present in the neutralizing compositions provided herein, in g / L, may be 0.10 or more, 1 .0 or more, 2.25 or more, 5.0 or more, 7.5 or more, 10.0 or more, 15.0 or more, or even 20.0 or more. The effective amount of at least one compound suitable for neutralizing phenols, hexachlorophene, formalin, and ethanol compounds present in the neutralizing compositions provided herein, in g / L, may 20.0 or more, 15.0 or less, 10.0 or less, 7.5 or less, 5.0 or less, 2.25 or less, 1 .0 or less, or even 0.10 or less.

[0036] According to one embodiment, the neutralizing compositions provided herein include at least one compound suitable for adjusting the overall pH of the neutralizing composition. The pH adjusting compound may be an acid or a base. Suitable bases include, but are not limited to, sodium carbonate, potassium hydroxide (KOH), sodium hydroxide (NaOH), or lithium hydroxide (LiOH). Suitable acids include, but are not limited, to sulfuric acid (H2SO4), hydrochloric acid (HCI), or trifluoroacetic acid (TFA). It will be appreciated by one of skill in the art that many suitable acids and (and requisite concentrations thereof) may be used to achieve a target overall pH of the neutralizing composition.

[0037] According to one embodiment, the neutralizing compositions provided herein include at least one compound suitable for aiding in the recovery of one or more microorganisms. According to one embodiment, the compound suitable for aiding in the recovery of one or more microorganisms includes a metal halide. According to one embodiment, the compound suitable for aiding in the recovery of one or more microorganisms includes a metal chloride. According to one embodiment, the compound suitable for aiding in the recovery of one or more microorganisms includes magnesium chloride (MgCI).

[0038] The at least one compound suitable for aiding in the recovery of one or more microorganisms may be present in the neutralizing composition at an effective amount. According to one embodiment, the effective amount of the at least one suitable compound for aiding in the recovery of one or more microorganisms present in the neutralizing compositions provided herein, in g / L, may be 0.01 or more, 0.05 or more, 0.10 or more, 0.20 or more, 0.25 or more, or even 0.30 or more. According to one embodiment, the effective amount of the at least one compound suitable for aiding in the recovery of one or more microorganisms present in theneutralizing compositions provided herein, in g / L, may be 0.30 or less, 0.25 or less, 0.20 or less, 0.10 or less, 0.05 or less, or even 0.01 or less.

[0039] According to one embodiment, the neutralizing compositions provided herein include at least one compound suitable for reducing foam (antifoam compound) in the neutralizing composition. The at least one antifoam compound also aids in reducing excessive bubbling and allows for vortexing without excessive bubbling which may cause volume loss during transfer back to a vial as described herein. The at least one suitable compound for foam reduction includes antifoam powder.

[0040] The at least one compound suitable for reducing foam in the neutralizing composition may be present in the neutralizing composition at an effective amount. According to one embodiment, the effective amount of the at least one compound suitable for reducing foam in the neutralizing composition, in g / L, may be 0.01 or more, 0.05 or more, 0.10 or more, 0.20 or more, 0.25 or more, 0.30 or more, or even 0.35 or more. According to one embodiment, the effective amount of the at least one compound suitable for reducing foam in the neutralizing composition, in g / L, may be 0.35 or less, 0.30 or less, 0.25 or less, 0.20 or less, 0.10 or less, 0.05 or less, or even 0.01 or less.

[0041] According to one embodiment, the neutralizing compositions provided herein include at least one buffer compound suitable for pH stabilization and aiding in the ability of the neutralizing composition to be stored long term with negligible degradation. The at least one buffer compound may be any buffer compound that resists changes in pH that prevent successful culturing or detection of microorganisms. The at least one buffer compound includes, but is not limited to, tris buffers, phosphate buffers, citrate buffers, acetate buffers. A particularly suitable buffer includes sodium phosphate dibasic.

[0042] The at least one buffer compound may be present in the neutralizing composition at an effective amount so as to provide sufficient stability to the neutralizing composition.According to one embodiment, the effective amount of buffer compound present in the neutralizing compositions provided herein, in g / L, may be 0.1 or more, 1 .0 or more, 1 .5 or more, 2.0 or more, 2.5 or more, or even 3.0 or more. The effective amount of buffer compound in the neutralizing compositions provided herein, in g / L, may be 3.0 or less, 2.5 or less, 2.0 or less, 1.5 or less, 1 .0 or less, or even 0.1 or less.

[0043] In any of the foregoing embodiments, the neutralizing composition does not include a primary amine.

[0044] In any of the foregoing embodiments, the neutralizing composition may be in the form of a solution or dispersion and has sufficient water or other suitable solvent or diluent todissolve or disperse all of the components described herein. In any embodiment, the neutralizing composition may have a pH of from about 6 to about 8. According to one embodiment, the neutralizing composition may have a pH of from about 6.5 to about 7.5.Related Methods, Vials and Kits

[0045] The neutralizing compositions provided herein may be utilized in methods that neutralize antimicrobial agents and allow growth of microorganisms that would otherwise be inhibited by the presence of antimicrobial agents. The methods provided herein result in the neutralization of antimicrobial compounds such that the antimicrobial compounds do not interfere with assays that may be used to detect or enumerate microorganisms after culturing.

[0046] A method of neutralizing at least one antimicrobial compound in a culture media provided. The method of neutralizing at least one antimicrobial compound includes the step of introducing an effective amount neutralizing composition to a culture media. The culture media is suitable for supporting growth of at least one microorganism. The culture media may include or otherwise be spiked with a test sample. The test sample may be obtained by sampling a surface to remove a test sample that includes, or is suspected of including, at least one microorganism. The surface may be sampled by using traditional techniques such as by using a dry sponge.

[0047] A method of culturing for one or more microorganisms is also provided. The method of culturing for one or more microorganisms includes the step of sampling a surface to remove a sample of the one or more microorganisms from the surface to form a test sample. The method of culturing for one or more microorganisms also includes the step of mixing the test sample with culture media to form a test sample composition. The method of culturing for one or more microorganisms also includes the step of contacting the test sample composition with a neutralizing composition as described herein so as to allow at least some of the one or more microorganisms to multiply within the growth media. The amount of neutralizing composition utilized may vary depending on the size of the vial and downstream assay device.

[0048] Vials for downstream detection assays are provided. The vials provided herein may be particularly suitable for use with devices for rapid detection of microorganisms. The vials may be sterile and include a growth medium. The vials may also include a neutralizing composition as provided herein or such neutralizing composition may be introduced separately to the vial. The vials may also include an agar plug that is separate from any test sample introduced to the vial. The agar plug includes those plugs set forth in U.S. Patent No. 9,850,457. The vials may be ready for inoculation with test sample and introduction to asuitable device for microorganism detection. The vials may be substantially transparent so as to allow light to pass through the vial. According to one embodiment, the vial is particularly suited to be used in conjunction with rapid microorganism detection devices that optically monitor growth of microorganisms present in a test sample based on pH changes, reagent color / fluorescence changes, or biochemical reactions. Such devices include, for example, the Soleris® rapid microbiological systems available from Neogen Corp.

[0049] A kit for detecting microorganisms in a test sample is provided. According to one embodiment, the kit may include a sealable specimen container, having an internal chamber comprising a culture or growth medium and in which a test sample, for which the presence or absence of a microorganism is to be determined, may be cultured. The sealable specimen container may be a vial as described herein.

[0050] According to one embodiment, the kit may further include at least one neutralizing composition as provide herein. The neutralizing composition may be added to the culture medium contained in the vial in an amount that is effective for neutralizing one or more antimicrobial compounds that may be present in a test sample that is introduced to the vial.EXAMPLESExample 1 Microorganism Growth Enhancement - Magnesium Chloride

[0051] Testing was conducted to observe the impact of growth enhancers (e.g., Magnesium Chloride) on Staphylococcus aureus when added to letheen broth. The letheen broth utilized in this Example was a liquid medium recommended for use in qualitative procedures for testing quaternary ammonium compounds for antimicrobial activity. The letheen broth included meat peptone, beef extract, Polysorbate 80 (polyoxyethylene sorbitan monooleate), sodium chloride, lecithin, and demineralized water.

[0052] The testing was conducted and modeled after EPA Standard Operating Procedure (SOP) MB-17-04 (Standard Operating Procedure for Neutralization Confirmation Procedure for Products Evaluated with the AOAC Use Dilution Method (UDM), the AOAC Germicidal Spray Products as Disinfectants Test (GSPT), and the Disinfectant Towelette Test (DTT)).Particularly, Staphylococcus aureus derived from ATCC® 6538 (Sa 6538) was inoculated at 18 CFU / mL into Soleris® vials containing 9 mL of various neutralizing compositions. During test incubation, the rapid microorganism detection device (Soleris® device) optically monitored for color change in an agar plug by plotting optical units at each reading. The optical unit is relativeto the color of the agar plug. Detection occurred when the rapid microorganism detection device observed an optical unit change above a test threshold and was reported as a detection time in hours. If no change in optical unit occurred, a reporting of “No Detection” was given.

[0053] Figure 1 sets forth a table that illustrates detection times when various levels of sodium bisulfite, sodium thiosulfate, sodium thioglycollate, lecithin, sodium chloride, magnesium chloride, and histidine are added to letheen broth. Even with comparable starting optical units, media version A8 was shown to exhibit significantly faster growth and recovery of Sa 6538 while AO exhibited slower growth. This result indicated that the addition of magnesium chloride provided microorganism growth enhancement.

[0054] The present disclosure enables one of skill in the relevant art to make and use the inventions provided herein in accordance with multiple and varied embodiments. Various alterations, modifications, and improvements of the present disclosure that readily occur to those skilled in the art, including certain alterations, modifications, substitutions, and improvements are also part of this disclosure. Accordingly, the foregoing description are by way of example to illustrate the discoveries provided herein. Furthermore, the foregoing Description and Examples are exemplary of the present invention and not limiting thereof. The scope of the invention is therefore set out in the appended claims.

[0055] Although specific embodiments of the present disclosure are herein illustrated and described in detail, the disclosure is not limited thereto. The above detailed descriptions are provided as exemplary of the present disclosure and should not be construed as constituting any limitation of the disclosure. Modifications will be obvious to those skilled in the art, and all modifications that do not depart from the spirit of the disclosure are intended to be included with the scope of the appended claims.

Claims

CLAIMS1 . A neutralizing composition comprising: at least one compound capable of neutralizing at least one antimicrobial compound; and at least one compound suitable for aiding in the recovery of one or more microorganisms.

2. The neutralizing composition of claim 1 , wherein the at least one compound capable of neutralizing at least one antimicrobial compound comprises a polyethylene sorbitol ester.

3. The neutralizing composition of claim 2, wherein the polyethylene sorbitol ester includes polyoxyethylene sorbitan monooleate.

4. The neutralizing composition of claim 1 , wherein the at least one compound capable of neutralizing at least one antimicrobial compound comprises lecithin.

5. The neutralizing composition of claim 1 , wherein the at least one compound capable of neutralizing at least one antimicrobial compound comprises casein peptone type 1 , meat peptone, beef extract, or any combination thereof.

6. The neutralizing composition of claim 1 , wherein the at least one compound capable of neutralizing at least one antimicrobial compound comprises L-histidine, sodium thiosulfate, or a combination thereof.

7. The neutralizing composition of claim 1 , wherein the at least one compound capable of neutralizing at least one antimicrobial compound comprises sodium bisulfite.

8. The neutralizing composition of claim 1 , wherein the at least one compound capable of neutralizing at least one antimicrobial compound comprises sodium carbonate.

9. The neutralizing composition of claim 1 , wherein the at least one compound suitable for aiding in the recovery of one or more microorganisms comprises a metal halide.

10. The neutralizing composition of claim 1 , wherein the at least one compound suitable for aiding in the recovery of one or more microorganisms comprises magnesium chloride.11 . The neutralizing composition of claim 1 , further comprising at least one yeast extract.

12. The neutralizing composition of claim 1 , further comprising sodium phosphate dibasic.

13. The neutralizing composition of claim 1 , further comprising at least one antifoaming compound.

14. The neutralizing composition of claim 13, wherein the at least one antifoaming compound is antifoam powder.

15. The neutralizing composition of claim 1 , wherein the neutralizing composition is shelf stable at ambient conditions for at least one week.

16. The neutralizing composition of claim 1 , having a pH of from about 6 to about 8.

17. A neutralizing composition comprising: about 5.0 g / L to about 15 g / L of lecithin; about 10.0 g / L to about 20.0 g / L of polyoxyethylene sorbitan monooleate; about 0.1 g / L to about 0.5 g / L of at least one antifoaming compound; and about 0.1 g / L to about 0.5 g / L of magnesium chloride.

18. The neutralizing composition of claim 17, further comprising peptone type 1 , meat peptone, beef extract, or any combination thereof.

19. The neutralizing composition of claim 17, further comprising L-histidine, sodium thiosulfate, or a combination thereof.

20. The neutralizing composition of claim 17, further comprising sodium bisulfite.21 . The neutralizing composition of claim 17, further comprising sodium carbonate.

22. The neutralizing composition of claim 17, further comprising at least one yeast extract.

23. The neutralizing composition of claim 17, further comprising sodium phosphate dibasic.

24. The neutralizing composition of claim 17, wherein the at least one antifoaming compound is antifoam powder.

25. The neutralizing composition of claim 17, wherein the neutralizing composition is shelf stable at ambient conditions for at least one week.

26. The neutralizing composition of claim 17, having a pH of from about 6 to about 8.

27. A vial comprising a neutralizing composition according to any one of claims 1 -26.

28. A method of neutralizing at least one antimicrobial compound in a culture medium comprising: introducing an effective amount neutralizing composition according to any one of claims 1 -26 to a culture media, the culture medium suitable for supporting growth of at least one microorganism, wherein the neutralizing composition neutralizing at least one antimicrobial compound in the culture medium.

29. A method of culturing for one or more microorganisms comprising: sampling a surface to remove a sample of the one or more microorganisms from the surface to form a test sample; mixing the test sample with culture medium to form a test sample composition; and contacting the test sample composition with a neutralizing composition according to any one of claims 1-26 so as to allow the one or more microorganisms to multiply within the culture medium.

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