U.S. patent application number 14/962128 was filed with the patent office on 2016-03-31 for antimicrobial compositions and methods of use thereof.
This patent application is currently assigned to SMARTWASH SOLUTIONS, LLC. The applicant listed for this patent is SMARTWASH SOLUTIONS, LLC. Invention is credited to Kel Eugene LEMONS.
Application Number | 20160088846 14/962128 |
Document ID | / |
Family ID | 40899881 |
Filed Date | 2016-03-31 |
United States Patent
Application |
20160088846 |
Kind Code |
A1 |
LEMONS; Kel Eugene |
March 31, 2016 |
ANTIMICROBIAL COMPOSITIONS AND METHODS OF USE THEREOF
Abstract
Aspects relate to an aqueous composition for use on produce. The
aqueous composition includes an acid, an organic diol, and an
oxidizing agent in the form of chlorine, wherein the aqueous
composition has acidic pH and has antimicrobial activity.
Inventors: |
LEMONS; Kel Eugene;
(Placerville, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
SMARTWASH SOLUTIONS, LLC |
Salinas |
CA |
US |
|
|
Assignee: |
SMARTWASH SOLUTIONS, LLC
Salinas
CA
|
Family ID: |
40899881 |
Appl. No.: |
14/962128 |
Filed: |
December 8, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12243460 |
Oct 1, 2008 |
9259006 |
|
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14962128 |
|
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61024558 |
Jan 30, 2008 |
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Current U.S.
Class: |
424/605 ; 134/26;
427/384 |
Current CPC
Class: |
A01N 37/36 20130101;
A61K 33/40 20130101; A01N 59/00 20130101; A61K 33/20 20130101; A01N
3/00 20130101; A01N 31/14 20130101; A01N 31/02 20130101; A01N
2300/00 20130101; A01N 31/14 20130101; A01N 41/02 20130101; A01N
59/00 20130101; A01N 59/26 20130101; A01N 37/36 20130101; A01N
31/02 20130101; A01N 2300/00 20130101; A01N 31/14 20130101; A01N
31/02 20130101; A01N 31/14 20130101; A01N 41/02 20130101; A01N
2300/00 20130101; A01N 41/02 20130101; A01N 41/02 20130101; A01N
59/00 20130101; A01N 31/02 20130101; A01N 2300/00 20130101; A61K
31/047 20130101; A01N 59/00 20130101; A61K 33/42 20130101; A23L
3/3481 20130101; A01N 37/02 20130101; A01N 37/36 20130101; A23L
3/3463 20130101; A01N 59/26 20130101; A01N 59/26 20130101; A23V
2002/00 20130101; A01N 37/02 20130101; A01N 59/00 20130101; A01N
37/02 20130101; A61L 2/18 20130101; B08B 3/08 20130101; A01N 37/36
20130101; A01N 59/00 20130101; A61L 2/186 20130101; A01N 59/26
20130101; A01N 31/02 20130101; A01N 37/02 20130101 |
International
Class: |
A01N 59/26 20060101
A01N059/26; A01N 59/00 20060101 A01N059/00; A01N 3/00 20060101
A01N003/00; A61L 2/18 20060101 A61L002/18; B08B 3/08 20060101
B08B003/08; A61K 33/40 20060101 A61K033/40; A61K 33/20 20060101
A61K033/20; A61K 31/047 20060101 A61K031/047; A23L 3/3481 20060101
A23L003/3481; A23L 3/3463 20060101 A23L003/3463; A01N 31/02
20060101 A01N031/02; A61K 33/42 20060101 A61K033/42 |
Claims
1. An aqueous composition for use on produce, the aqueous
composition comprising: an acid; an organic diol; and an oxidizing
agent in the form of chlorine, wherein the aqueous composition has
acidic pH and has antimicrobial activity.
2. The composition of claim 1, wherein the amount of said acid
alone and said organic diol alone has lower antimicrobial activity
compared to antimicrobial activity of a combination of said acid,
said organic diol, and chlorine.
3. The composition of claim 1, wherein said composition comprises
from 0.001 wgt. % to 40 wgt. % of said acid.
4. The composition of claim 1, wherein said composition comprises
from 0.001 wgt. % to 5 wgt. % of said organic diol.
5. The composition of claim 1, wherein said composition comprises
from 0.001 wgt. % to 30 wgt. % of said oxidizing agent.
6. The composition of claim 1, wherein said composition further
comprises a surfactant.
7. The composition of claim 1, wherein said composition comprises
from 0.001 wgt. % to 0.1 wgt. % of said surfactant.
8. The composition of claim 1, wherein said composition is at a
temperature from below said composition's freezing temperature to
120 degrees Celsius.
9. The composition of claim 1, wherein antimicrobial activity of
(a) said acid, (b) said organic diol, (c) said oxidizing agent, (d)
a combination of said acid and of said organic diol, (e) a
combination of said acid and of said oxidizing agent, and (f) a
combination of said organic diol and of said oxidizing agent, is
lower than antimicrobial activity of a combination of said acid, of
said organic diol, and of said oxidizing agent.
10. An agricultural product having a surface film that comprises:
an acid that does not contain an --NH group and an --NH.sub.2
group; an organic diol; and an oxidizing agent in the form of
chlorine.
11. A method for making an aqueous antimicrobial composition, the
method comprising: providing an acid that does not contain an --NH
group and an --NH.sub.2 group; providing an organic diol; providing
an oxidizing agent in the form of chlorine; and mixing said acid,
said diol, and said oxidizing agent in water to produce an aqueous
antimicrobial composition.
12. The method of claim 11, further comprising: mixing a surfactant
in said aqueous antimicrobial composition.
13. A method for making an aqueous antimicrobial composition, the
method comprising: providing an acid; providing an organic diol;
providing an oxidizing agent in the form of chlorine; mixing said
acid and said diol in water to produce a first mixture; and mixing
said oxidizing agent with said first mixture to produce an aqueous
antimicrobial composition.
14. The method of claim 13, further comprising: mixing a surfactant
in said aqueous antimicrobial composition.
15. A method for reducing the number of microbes on a surface, the
method comprising: providing a first surface comprising a first
number of microbes providing an antimicrobially effective amount of
an aqueous antimicrobial composition that includes an acid, an
organic diol, and an oxidizing agent in the form of chlorine; and
contacting said first surface with said aqueous antimicrobial
composition under conditions that produce a contacted surface
comprising a reduced number of said microbes compared to said first
number of said microbes on said first surface in the absence of
said contacting.
16. The method of claim 15, wherein said first surface comprises: a
pathogenic microbe and a non-pathogenic microbe, and said reduced
number of said microbes comprises a greater reduction in the number
of said pathogenic microbe than in the number of said
non-pathogenic microbe.
17. The method of claim 15, further comprising: drying said
contacted surface to produce a dried surface comprising one or more
of said acid, said organic diol, and said oxidizing agent.
18. The method of claim 15, further comprising: washing said
contacted surface under conditions that reduce the amount of one or
more of said acid, of said organic diol, and of said oxidizing
agent on said contacted surface.
19. A method for reducing discoloration of an agricultural product,
comprising: providing an agricultural product having a first
surface; providing an anti-discoloration effective amount of an
aqueous antimicrobial composition that includes an acid, an organic
diol, and an oxidizing agent in the form of chlorine; and
contacting said agricultural product with said aqueous
antimicrobial composition under conditions that produce a contacted
surface having reduced discoloration compared to discoloration of
said first surface in the absence of said contacting.
Description
[0001] This application is a continuation of U.S. patent
application Ser. No. 12/243,460 filed on Oct. 1, 2008 that
published as U.S. Patent Application Publication No. 2009/0192231.
The contents of the priority application Ser. No. 12/243,460 is
hereby incorporated by reference in its entirety.
FIELD OF THE INVENTION
[0002] The invention provides antimicrobial compositions comprising
one or more acid and one or more organic diol. In one embodiment,
the invention's compositions have an acidic pH. The compositions
may optionally further contain one or more oxidizing agent
(including stabilized oxidizing agent and/or unstabilized oxidizing
agent), and/or one or more surfactant. In particular embodiments,
the acid lacks one or both of --NH group and --NH.sub.2 group. The
invention's compositions are useful in all stages of preparation of
agricultural products, in hospitals, and in commercial and
household applications.
BACKGROUND OF THE INVENTION
[0003] Microbial contamination of food, in hospitals, in animals,
as well as in commercial and residential buildings continues to be
a problem. In particular, pathogenic microbial contamination of
fruits and vegetables has been on the rise, including contamination
of fruits and vegetables in the field, during harvesting, transport
and/or processing, by pathogenic microbes from animals, humans,
organic fertilizer and/or water supply. Bacteria are estimated to
cause some 24 million cases of diarrheal disease annually in the
U.S. Over 40,000 cases of contamination by Salmonella were reported
from 1983 to 1987 in laboratory surveillance data.
[0004] Several compositions and methods are available for reducing
microbial contamination (e.g., Bailey et al., U.S. Pat. No.
T964,007 and U.S. Pat. No. 4,074,058; Faergemann et al.,
WO/89/12469; Andrews et al., U.S. Pat. No. 5,490,992; Estrada, U.S.
Pat. No. 5,599,571; Andrews, U.S. Pat. No. 5,569,461; Bautista et
al., U.S. Pat. No. 6,045,846; Conners et al., U.S. Pat. No.
6,086,833; Bender et al., U.S. Pat. No. 6,287,617; Koefod et al.,
U.S. Pat. No. 7,090,882; Howarth et al., U.S. Pat. No. 7,182,966;
Shane et al., U.S. Patent Application US 2002/0134317; and Hilgren
et al., U.S. Patent Application US 2007/0098751), including those
that contain oxidizers. However, prior art compositions that
contain oxidizers continue to show low antimicrobial activity in
the presence of organic contaminants, and especially at the lower
temperatures used for processing, storing and transporting
agricultural products. Also, the prior art's compositions have the
additional problems of increased foaming and of the oxidizer
gassing out of the solution due to bio-load and/or too low or too
high pH, thus adversely impacting active ingredients and worker
safety. Furthermore, higher levels of oxidizers are required to
reduce bio-load and turbidity of the solutions used for
antimicrobial treatment, which raises safety concerns when
ingesting products that are treated with those compositions. In
addition, the presence of plant material in the prior art solutions
reduces the longevity of the oxidizer, thus reducing its
antimicrobial activity and increasing the cost of production
because of the need to replenish the oxidizer often.
[0005] Thus, there remains a need for improved compositions and
methods for reducing microbial contamination.
SUMMARY
[0006] In accordance with one or more embodiments, an aqueous
composition for use on produce is provided. The aqueous composition
includes an acid, an organic diol, and an oxidizing agent in the
form of chlorine, wherein the aqueous composition has acidic pH and
has antimicrobial activity.
[0007] In accordance with another embodiment, the amount of said
acid alone and said organic diol alone has lower antimicrobial
activity compared to antimicrobial activity of a combination of
said acid, said organic diol, and chlorine.
[0008] In accordance with another embodiment, said composition
includes from 0.001 wgt. % to 40 wgt. % of said acid. In accordance
with another embodiment, said composition includes from 0.001 wgt.
% to 5 wgt. % of said organic diol. In accordance with another
embodiment, said composition includes from 0.001 wgt. % to 30 wgt.
% of said oxidizing agent.
[0009] In accordance with another embodiment, said composition
further includes a surfactant. In accordance with another
embodiment, said composition includes from 0.001 wgt. % to 0.1 wgt.
% of said surfactant.
[0010] In accordance with another embodiment, said composition is
at a temperature from below said composition's freezing temperature
to 120 degrees Celsius.
[0011] In accordance with another embodiment, antimicrobial
activity of (a) said acid, (b) said organic diol, (c) said
oxidizing agent, (d) a combination of said acid and of said organic
diol, (e) a combination of said acid and of said oxidizing agent,
and (f) a combination of said organic diol and of said oxidizing
agent, is lower than antimicrobial activity of a combination of
said acid, of said organic diol, and of said oxidizing agent.
[0012] In accordance with one or more embodiments, an agricultural
product having a surface film is provided. The agricultural product
includes an acid that does not contain an --NH group and an
--NH.sub.2 group, an organic diol, and an oxidizing agent in the
form of chlorine.
[0013] In accordance with one or more embodiments, a method for
making an aqueous antimicrobial composition is provided. The method
includes providing an acid that does not contain an --NH group and
an --NH.sub.2 group, providing an organic diol, providing an
oxidizing agent in the form of chlorine, and mixing said acid, said
diol, and said oxidizing agent in water to produce an aqueous
antimicrobial composition.
[0014] In accordance with another embodiment, the method further
includes mixing a surfactant in said aqueous antimicrobial
composition.
[0015] In accordance with one or more embodiments, a method for
making an aqueous antimicrobial composition is provided. The method
includes providing an acid, providing an organic diol, providing an
oxidizing agent in the form of chlorine, mixing said acid and said
diol in water to produce a first mixture, and mixing said oxidizing
agent with said first mixture to produce an aqueous antimicrobial
composition.
[0016] In accordance with another embodiment, the method, further
includes mixing a surfactant in said aqueous antimicrobial
composition.
[0017] In accordance with one or more embodiments, a method for
reducing the number of microbes on a surface is provided. The
method includes providing a first surface including a first number
of microbes providing an antimicrobially effective amount of an
aqueous antimicrobial composition that includes an acid, an organic
diol, and an oxidizing agent in the form of chlorine, and
contacting said first surface with said aqueous antimicrobial
composition under conditions that produce a contacted surface
including a reduced number of said microbes compared to said first
number of said microbes on said first surface in the absence of
said contacting.
[0018] In accordance with another embodiment, said first surface
includes a pathogenic microbe and a non-pathogenic microbe, and
said reduced number of said microbes includes a greater reduction
in the number of said pathogenic microbe than in the number of said
non-pathogenic microbe.
[0019] In accordance with another embodiment, the method further
includes drying said contacted surface to produce a dried surface
including one or more of said acid, said organic diol, and said
oxidizing agent.
[0020] In accordance with another embodiment, the method further
includes washing said contacted surface under conditions that
reduce the amount of one or more of said acid, of said organic
diol, and of said oxidizing agent on said contacted surface.
[0021] In accordance with one or more embodiments, a method for
reducing discoloration of an agricultural product is provided. The
method includes providing an agricultural product having a first
surface, providing an anti-discoloration effective amount of an
aqueous antimicrobial composition that includes an acid, an organic
diol, and an oxidizing agent in the form of chlorine, and
contacting said agricultural product with said aqueous
antimicrobial composition under conditions that produce a contacted
surface having reduced discoloration compared to discoloration of
said first surface in the absence of said contacting.
[0022] Additional features and advantages are realized through the
techniques of the present invention. Other embodiments and aspects
of the invention are described in detail herein and are considered
a part of the claimed invention. For a better understanding of the
invention with the advantages and the features, refer to the
description and to the drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0023] The subject matter which is regarded as the invention is
particularly pointed out and distinctly claimed in the claims at
the conclusion of the specification. The forgoing and other
features, and advantages of the invention are apparent from the
following detailed description taken in conjunction with the
accompanying drawings in which:
[0024] FIG. 1 shows the level of hypochlorous acid in solution, as
determined by the DPD assay. (A) Composition "x1a" containing
ortho-phosphoric acid 0.03 wgt. %, propylene glycol 0.02 wgt. %,
hypochlorous acid 0.005 wgt. % at pH 4. (B) Chlorine solution
containing ortho-phosphoric acid 0.001 wgt. % and hypochlorous acid
0.005 wgt. % at pH 7.
DETAILED DESCRIPTION
[0025] Definitions
[0026] To facilitate understanding of the invention, a number of
terms are defined below.
[0027] The term "wgt. %" refers to the relative weight of two
compounds when expressed as a percentage. For example, a solution
containing 10 wgt. % of ortho-phosphoric acid propylene glycol
dissolved in water refers to a solution prepared by diluting 10
grams of ortho-phosphoric acidpropylene glycol with water to a
final weight of 100 grams.
[0028] "Logarithm" and "log" of a number, such as when used in
reference to a concentration, cell number, etc., interchangeably
refer to the power or exponent to which a base of 10 must be raised
in order to produce the number. For example, 6-log means 1,000,000.
Thus, a 3-log reduction means a 1,000 fold reduction.
[0029] "Aqueous" solution or composition refers to a solution or
composition in which the solvent is water.
[0030] An "organic" chemical compound (e.g., organic acid, organic
diol, organic oxidizer, organic solvent, etc.) is a chemical
compound that contains carbon. In contrast, an "inorganic" chemical
compound (e.g., inorganic acid, inorganic oxidizer, inorganic
solvent, etc.) is a chemical compound that does not contain
carbon.
[0031] "Microbe" and "microorganism" interchangeably refer to an
organism that is too small to be seen by the naked human eye.
Microbes include prokaryotes (such as bacteria, and virus
(including bacteriophage), etc.), and eukaryotes (such as protists
(including nematodes), animals, fungi, and plants). Microbes may be
pathogenic or non-pathogenic.
[0032] "Pathogen" refers to an organism that causes a disease in a
multicellular animal. In contrast, a "non-pathogen" is an organism
that does not cause disease in an animal. "Animal" includes mammals
(e.g., humans, non-human primates, murines, ovines, bovines,
ruminants, lagomorphs, porcines, caprines, equines, canines,
felines, ayes, etc.), avians (e.g., chicken), amphibians (e.g.,
Xenopus), reptiles, etc.
[0033] The term "bacteria" refers to all prokaryotic organisms,
including those within all of the phyla in the Kingdom Procaryotae.
Bacteria include Mycoplasma, Chlamydia, Actinomyces, Streptomyces,
and Rickettsia. Forms of bacteria include cocci, bacilli,
spirochetes, spheroplasts, protoplasts, etc.. Bacteria may be
Gram-negative or Gram-positive. "Gram-negative" and "Gram-positive"
refer to staining patterns with the Gram-staining process that is
well known in the art (Finegold and Martin, Diagnostic
Microbiology, 6th Ed. (1982), CV Mosby St. Louis, pp 13-15).
"Gram-positive bacteria" are bacteria that retain the primary dye
used in the Gram-stain, causing the stained cells to appear dark
blue to purple under the microscope. "Gram-negative bacteria" do
not retain the primary dye used in the Gram-stain, but are stained
by the counterstain. Thus, Gram-negative bacteria appear red.
[0034] "Gram-negative bacteria" include the proteobacteria,
exemplified by Escherichia coli, Salmonella, and other
Enterobacteriaceae, Pseudomonas, Moraxella, Helicobacter,
Stenotrophomonas, Bdellovibrio, acetic acid bacteria, Legionella,
alpha-proteobacteria such as Wolbachia and many others.
Gram-negative bacteria include bacilli that cause respiratory
problems (such as Hemophilus influenzae, Klebsiella pneumoniae,
Legionella pneumophila, Pseudomonas aeruginosa), urinary problems
(Escherichia coli, Proteus mirabilis, Enterobacter cloacae,
Serratia marcescens), and gastrointestinal problems (Helicobacter
pylori, Salmonella enteritidis, Salmonella typhi and
Campylobacter). Gram-negative bacteria associated with nosocomial
infections include Acinetobacter baumanii, which cause bacteremia,
secondary meningitis, and ventilator-associated pneumonia in
intensive care units of hospital establishments.
[0035] "Gram-positive bacteria" include the phylum Firmicutes
(exemplified by the genera Bacillus, Listeria, Staphylococcus,
Streptococcus, Enterococcus, and Clostridium), and the Mollicutes
(exemplified by Mycoplasma that lack cell walls and cannot be Gram
stained). Listeria monocytogenes is a Gram-positive bacteria that
is of particular concern in many ready-to-eat meat, poultry,
seafood, and dairy processing chill brine applications.
[0036] The term "virus" refers to obligate, ultramicroscopic,
intracellular parasites incapable of autonomous replication (i.e.,
replication requires the use of the host cell's machinery). Viruses
are exemplified by, but not limited to, rhinovirus, human papilloma
virus, human immunodeficiency virus, hepatitis virus, Newcastle
disease virus, cardiovirus, corticoviridae, cystoviridae,
epstein-barr virus, filoviridae, hepadnviridae, herpes virus,
influenza virus, inoviridae, iridoviridae, metapneumovirus,
orthomyxoviridae, papovavirus, paramyxoviridae, parvoviridae,
polydnaviridae, poxyviridae, reoviridae, rhabdoviridae, semliki
forest virus, tetraviridae, toroviridae, vaccinia virus, and
vesicular stomatitis virus.
[0037] A "fungus" is a eukaryotic organism that is a member of the
kingdom Fungi. Fungi are heterotrophic organisms possessing a
chitinous cell wall. The majority of species grow as multicellular
filaments called hyphae forming a mycelium. Some fungal species
also grow as single cells. Fungi include "mold," which are
microscopic fungi that grow in the form of multicellular filaments,
called hyphae. Molds include dermatophytes (i.e., pathogenic fungus
that infects the skin) exemplified by the genera Epidermophyton,
Microsporum and Trichophyton (such as Trichophyton rubrum and
Trichophyton mentagrophytes). Trichophyton can cause tinea pedis
(athlete's foot) and/or tinea cruris (jock itch). Plant fungi
include flagellated fungi and non-flagellated fungi.
[0038] Flagellated fungi include Plasmodiophoromycetes,
Chytridiomycota and Oomycetes. Plasmodiophoromycetes include
Plasmodiophora brassicae that causes club root of cabbage, and
Spongospora subterranean that causes powdery scab of potatoes.
Chytridiomycota include Olpidium spp. that infect pollen,
Physoderma maydis that causes brown spot of corn leaves, and
Synchytrium spp. that cause wart of potatoes. The Peronosporales
group is exemplified by the late blight of potato fungus
Phytophthora infestans, Peronospora, Bremia, Plasmopara and others
that cause "downy mildews", the "damping off' fungi, Pythium spp.,
and the white rust fungi, Albugo spp.
[0039] Non-flagellated Fungi include Zygomycota, Ascomycota,
Deuteromycetes and Basidiomycetes. Zygomycota include the Mucorales
that are exemplified by members of the bread mold genus Rhizopus,
and Hoanephora that causes blossom blight and decay of squash.
Ascomycota include Taphrina deformans that causes peach leaf curl,
and Nematospora that causes seed decay and root rot on cotton,
Plectomycetes such as Ophiostoma (Ceratocystis), and O. ulmi that
causes Dutch elm disease, Pyrenomycetes such as powdery mildews of
the genus Erysiphe that is common on grasses, Phyllactinia on oaks
and other trees, and Uncinula on grapes and other shrubs,
Discomycetes such as Sclerotinia that causes stromatic rot of
vegetables, and Monilinia, the cause of brown rot of peaches, and
Loculoascomycetes such as Myriangiales that include Elsinoe species
that cause citrus scab, and Dothideales that include Capnodium
species that cause sooty molds of plants. Deuteromycetes include
Ascomycetes such as species of Alternaria, Bipolaris, Botrytis,
Cercospora, Diplodia, Dreschlera, Exerohilum, Fusarium, Phoma,
Phomopsis, Rhizoctonia, and Verticillium that cause molds, blights,
cankers, leaf spots, and root rots. Basidiomycetes include
Uredinales, Exobasidiales, and Aphyllophorales that cause rusts,
smuts, felt fungi, root rots, heart rots, and thread-blights.
[0040] A "bacteriophage" is a virus that infects bacteria.
Bacteriophages are classified in the families Myoviridae,
Siphoviridae, Podoviridae, Tectiviridae, Corticoviridae,
Lipothrixviridae, Plasmaviridae, Rudiviridae, Fuselloviridae,
Inoviridae, Microviridae, Leviviridae, and Cystoviridae.
[0041] "Nematode" and "roundworm" interchangeably refer to a
multicellular organism that is a member of the Phylum Nematoda.
Exemplary nematodes that are agricultural pests include corn pests
such as Belonolaimus (Sting Nematode), Criconemoides (Ring
nematode), Helicotylenchus (Spiral Nematode), Heterodera zeae (Corn
Cyst Nematode), Hoplolaimus (Lance Nematode), Xiphinema (Dagger
Nematode), Longidorus (Needle Nematode), Meloidogyne (Root-Knot
Nematode), Pratylenchus (Lesion Nematode), Paratrichodorus
(Stubby-Root Nematode), Tylenchorhynchus (Stunt Nematode); potato
pests such as Meloidogyne chitwoodi (Columbia Root-knot Nematode),
Globodera rostochiensis (Golden Nematode), Meloidogyne hapla
(Northern Root Knot Nematode), Ditylenchus destructor (Potato Rot
Nematode), Globodera pallida (Pale Potato Cyst Nematode); soybean
pests such as Heterodera glycines (Soybean cyst nematode) and
Belonolaimus spp. (Sting nematode); sugar beet pests such as
Heterodera schachtii (Sugar beet cyst nematode) and Nacobbus
aberrans (False root-knot nematode); turf pests such as
Belonolaimus species (Sting Nematode), Hoplolaimus galeatus (Lance
Nematode), Meloidogyne species (Root-knot Nematodes) and
Criconemoides species (Ring Nematode); trees and vines pests such
as Bursaphelenchus xylophilus (Pine wilt nematode), Tylenchulus
semipenetrans (Citrus nematode), Radopholus similis (Burrowing
nematode), Belonolaimus longicaudatus (Sting nematode), Xiphinema
americanum (Dagger nematode), Mesocriconema xenoplax (Ring
nematode), Meloidogyne hapla (Root-knot nematode), Tylenchorhynchus
spp. (Stunt nematode), Rotylenchulus spp. (Reniform nematode) and
Pratylenchus spp. (Lesion nematode); ornamentals and garden
vegetables pests such as Aphelenchoides spp. (Foliar nematodes),
Meloidogyne spp. (Root-knot nematodes), Ditylenchus dipsaci (Stem
and bulb nematode), and Belonolaimus longicaudatus (Sting
nematode).
[0042] The term "surface" refers to one or more of the faces of a
three-dimensional object, including surfaces that are visible to
the naked eye (e.g., agricultural products, inanimate objects,
etc.) and surfaces not visible to the naked eye (e.g., open stomata
and damaged cells on a plant leaf surface).
[0043] "Agricultural product" refers to plant material and/or
animal material that may be useful to man, as well as products
derived directly or indirectly therefrom (e.g., cut flowers, cooked
meat, etc.). Agricultural products include food products and
non-food products.
[0044] "Food product" refers to an agricultural product that is
"edible" (i.e., suitable for use as food) to at least one animal
(e.g., human, livestock, etc.). For example, the invention's
compositions may be edible when they are present as a film on a
food product and when the type and/or amount of the components are
generally recognized as safe (GRAS). Food products include
"pre-harvest" and "post-harvest" products. Food products are
exemplified by, but not limited to, fruits, vegetables, herbs,
seeds, nuts, meat, poultry, seafood, poultry eggs, etc. Food
products may be fresh or processed. "Fresh food product" refers to
a food product that has not been cooked or frozen (i.e., exposed to
temperatures higher or lower than room temperature, i.e., ambient
temperature). For example, a "fresh" food product includes, but is
not limited to a, raw food product, unprocessed food product, food
product that has not been heated above ambient temperature, food
product that has not been cooled below ambient temperature, food
product that does not contain a preservative, and/or irradiated
food product. A "fresh" food product may be "fresh frozen," "frozen
fresh," and "freshly frozen," which are terms that interchangeably
refer to a food product that is quickly frozen while still fresh. A
"fresh" food product may be blanched, i.e., briefly scalded, before
freezing to prevent nutrient breakdown. "Produce" is a fresh food
product derived from a plant (e.g., fruit, vegetable, herb, seed,
nut, leaf, stem, bulb). "Processed food product" refers to a food
product that has been manipulated by man including a product that
has been cut, chopped, sliced, peeled, ground, milled, irradiated,
frozen, cooked (e, g, blanched and/or pasteurized), homogenized,
germinated, washed, colored, waxed, hydro-cooled, refrigerated,
shelled, and/or had leaves, stems, or husks removed.
[0045] "Non-food product" refers to an agricultural product that is
not edible to at least one animal. Non-food products include cut
flowers, houseplants, animal hides, feathers, skin, shells,
etc.
[0046] "Inanimate object" refers to an object not endowed with life
or spirit such as equipment that is used for processing and/or
harvesting agricultural products, hospital instruments and
surfaces, surgical tools and apparatus, household items, buildings,
tunnels, underpasses, overpasses, floors, doors (frames, door
knobs, etc.), entrances, exits, ceilings, bathrooms and fixtures,
furniture, wood, Teflon.TM., ultra high molecular weight (UMHW)
polyethylene, ceramic tile, steel (e.g., stainless steel), cement,
glass, paint (e.g., acrylic paint), and reverse osmosis (RO)
membrane (i.e., semi permeable thin film composite membranes,
referred to as TFC or TFM).
[0047] "Skin" refers to the integument of an animal separated from
the body, with or without hair. The animal may be living or
non-living. Skin may be on the outside of the body, tissue on the
inside of the body, and/or tissue at or near openings on the body,
including openings made surgically.
[0048] "Film" refers to a thin covering, coating or layer. A film
may be "clear" i.e., transparent and/or translucent. Alternatively,
the film may be "opaque." Films may be visible to the human naked
eye or invisible to the human naked eye. A film may be "edible,"
i.e., is not toxic when ingested by an animal. To illustrate, an
edible film containing one or more of the invention's compositions
includes, without limitation, a film in which the components are
listed as GRAS and/or FCC. A film may be "adherent," i.e., is not
removed from a surface after rinsing with water for 1 minute. A
liquid film may appear dry to the naked human eye.
[0049] "Bio-film" is a film containing an aggregation of
microorganisms. Bio-films are characterized by structural
heterogeneity, genetic diversity, complex community interactions,
and an extracellular matrix of polymeric substances.
[0050] "Bio-load" and "biological load" refer to the waste output
of organisms, plant matter, soil, extracts, and any other
biological matter that is capable of changing the oxygenation rate
of a fluid, e.g., water.
[0051] The terms "reduce," "decrease," and grammatical equivalents
when in reference to a numerical value (e.g., number of cells,
concentration, etc.) of any molecule (e.g., acid, diol, oxidizing
agent, surfactant, etc.), and/or phenomenon (e.g., antimicrobial
activity, microbial contamination, cell division, cell viability,
discoloration, pinking, browning, etc.) in a first sample relative
to a second sample, mean that the quantity of the numerical value
in the first sample is lower than in the second sample by any
amount that is statistically significant using any art-accepted
statistical method of analysis. The reduction may be determined
objectively and/or subjectively. In one embodiment, the quantity of
the numerical value in the first sample is at least 10% lower than,
at least 25% lower than, at least 50% lower than, at least 75%
lower than, at least 90% lower than, at least 99% lower than,
and/or 100% lower than the quantity of the numerical value in the
second sample.
[0052] The terms "increase," and grammatical equivalents when in
reference to a numerical value (e.g., number of cells,
concentration, etc.) of any molecule (e.g., acid, diol, oxidizing
agent, surfactant, etc.), and/or phenomenon (e.g., antimicrobial
activity, microbial contamination, cell division, cell viability,
discoloration, pinking, browning, etc.) in a first sample relative
to a second sample, mean that the quantity of the numerical value
in the first sample is higher than in the second sample by any
amount that is statistically significant using any art-accepted
statistical method of analysis. The increase may be determined
objectively and/or subjectively. In one embodiment, the quantity of
the numerical value in the first sample is at least 10% greater
than, at least 25% greater than, at least 50% greater than, at
least 75% greater than,and/or at least 90% greater than, and/or at
least 100% greater than the quantity of the same molecule and/or
phenomenon in the second sample.
[0053] "Acidic pH" refers to a pH less than 7.0, as exemplified by,
but not limited to, a pH less than pH 6.5, less than pH 6.0, less
than pH 5.5, less than pH 5.0, less than pH 4.5, less than pH 4.0,
less than pH 3.5, less than pH 3.0, less than pH 2.5, less than pH
2.0, and less than pH 1.5. Thus, an acidic pH includes a pH from
2.0 to 6.0, from 2.0 to 5.0, from 2.0 to 4.0, and from 2.0 to
3.0.
[0054] Unless otherwise indicated, all numbers expressing
quantities of ingredients, temperatures, microbes, reaction
conditions, and so forth as used herein, are to be understood as
being modified in all instances by the term "about," which refers
to variation in the numerical quantity that can occur, for example,
through typical measuring and liquid handling procedures used for
making concentrates or using solutions in the real world; through
inadvertent error in these procedures; through differences in the
manufacture, source, or purity of the ingredients employed to make
the compositions or carry out the methods; and the like.
Accordingly, unless indicated to the contrary, the numerical
parameters herein are approximations that may vary depending upon
the desired properties sought to be obtained by the present
invention. Notwithstanding that the numerical ranges and parameters
describing the broad scope of the invention are approximations, the
numerical values in the specific examples are reported as precisely
as possible. Any numerical value, however, inherently contains
standard deviations that necessarily result from the errors found
in the numerical value's testing measurements.
[0055] The invention provides antimicrobial compositions comprising
one or more acid and one or more organic diol. In one embodiment,
the invention's compositions have an acidic pH. The compositions
may optionally further contain one or more oxidizing agent
(including stabilized oxidizing agent and/or unstabilized oxidizing
agent), and/or one or more surfactant. In particular embodiments,
the acid lacks one or both of --NH group and --NH2 group.
[0056] The invention's discovery of the invention's compositions
that contain a combination of an acid and organic diol was
premised, in part, on the surprising discovery of synergistic
antimicrobial activity of this combination, including at acidic
pH.
[0057] The invention's discovery of the invention's compositions
that contain a combination of an acid, organic diol and an oxidizer
was premised, in part, on the surprising discovery that adding an
organic diol to an oxidizer in solution resulted in the ability to
alter (increase or decrease) the pH of the solution independently
of the bio-load, while also providing synergistic antimicrobial
activity of the acid, organic diol and oxidizer.
[0058] In addition, inclusion of the organic diol unexpectedly
reduced the undesirable odor when an oxidizer is included in the
composition, particularly in the presence of bioload. This allowed,
in one embodiment, the further addition of acids to the solution,
which resulted in surprising synergistic antimicrobial activity by
the acid, organic diol, and oxidizing agent. Furthermore, the
inventor made the surprising observation that particular
embodiments of the invention's compositions were capable of 100%
reduction in microbial contamination at the chilling temperatures
(e.g., from 30.degree. F. to 40.degree. F.) that are routinely used
during standard processing steps for agricultural products, while
not resulting in adverse effects on the odor, texture, color and/or
appearance of the agricultural products. A further surprising
aspect of the invention's compositions is that they had a broad
spectrum of antimicrobial activity against bacterial,
bacteriophage, viral, fungal and nematode microbes, using a wide
variety of agricultural products and inanimate objects. Moreover,
the invention's compositions surprisingly reduced discoloration
(e.g., pinking and browning) of agricultural products. Yet another
surprising property of the invention's compositions is that they
showed a differential effect on pathogenic and non-pathogenic
microbes, by reducing the number of pathogenic bacteria to a
greater extent than the reduction of non-pathogenic bacteria and of
non-pathogenic bacteriophage.
[0059] The invention's compositions have several additional
advantages over antimicrobial compositions of the prior art. For
example, in one embodiment, the invention's compositions are clear
solutions, non-foaming during use, and continue to exhibit residual
antimicrobial activity after drying.
[0060] In addition, the invention's compositions in one embodiment
exhibit antimicrobial activity at acidic pH, thus advantageously
allowing, in one embodiment, chlorine salts to remain in
solution.
[0061] Moreover, after drying, the invention's compositions form a
very thin film on the surface (including cracks and open stomata)
of the treated products consisting of the composition's components.
This film is edible because of its low toxicity to higher life
forms, while continuing to exhibit antimicrobial activity, thus
eliminating the need, and environmental and monetary cost, for
rinsing off the invention's composition from the treated products
(e.g., agricultural products). Moreover, a film can be placed on
machines (or portions thereof such as cutting blades) that process
agricultural products.
[0062] In addition, in one embodiment, the invention's compositions
may be applied pre-harvest and post harvest of agricultural
products, thus advantageously extending the period of exposure to
antimicrobial activity, and resulting in increased shelf life.
[0063] Another advantage of the invention's compositions is that
their antimicrobial activity is resistant to particulate loads,
increased turbidity, and plant fluids. They also decrease turbidity
in the solution, allow for lower processing temperatures (one to
two of), and decrease the incidence of ice crystal formation on the
treated materials at refrigerated temperatures, thereby preserving
surface quality.
[0064] A further advantage is that the invention's compositions are
non-irritating when applied to skin as antiseptic even after 12
hours of contact, thus making them useful in medical applications.
In one embodiment, compositions of the present invention can be
applied directly to the skin. In another embodiment, the present
invention contemplates surgical dressings, bandages and surgical
tapes comprising one or more of the compositions set forth herein
(see Tables below). Illustrative examples of such dressings and
tapes include, but are not limited to, sheets of material, surgical
swabs, gauze pads, closure strips, compress bandages, surgical
tape, and the like. In a further embodiment, medical devices are
contacted (and preferably coated, e.g., coated with a film) with
compositions set forth herein (see Tables below). Such devices can
be used both outside and in the body, including implantable medical
devices (e.g., staples, shunts, stents, stitches, tubing, and the
like). In one embodiment, teeth or dental products are contacted
with compositions set forth herein. In yet another embodiment,
contact lenses and related eye care articles are contacted with
compositions set forth herein.
[0065] An additional advantage is that the invention's compositions
may be made as concentrates and used in situ and ex situ. Yet
another advantage is that they may be readily applied by standard
methods such as spraying, dipping, wiping, etc., thus avoiding the
need for costly modifications to existing commercial
applications.
[0066] Thus, the invention's compositions are useful for
antimicrobial applications in a variety of settings, such as before
harvesting and/or at the point of harvesting and/or after
harvesting of agricultural products (including food products,
ornamental plants, etc.), at food production facilities including
the disinfection of food processing equipment and harvesting
equipment, in hospitals, offices, residences, etc. For example, the
invention's compositions may be used pre-harvest for microbe
reduction on living plants. Also, ice made of the invention's
compositions may be used for cold storage of shellfish, fruits,
vegetables and meats. The compositions also may be used to reduce
undesirable odors from bacterial breakdown of food materials.
Further utilities include reduction of microbial infection of skin,
such as athlete's foot fungus "tinea pedis" (Trichophyton).
[0067] In addition, the invention's compositions may be used for
rendering any type of surface resistant to mold, for cleaning
and/or pre-treating such surfaces, and for rendering surfaces
contaminated with potentially or actually hazardous microbes such
as, but not limited to, mold spores, bacteria, viruses, protozoa
and/or any biological warfare agents, safe to handle and/or safe to
dispose of
[0068] Additional advantages, surprising results and attributes of
the invention are further described below under (A) Compositions
and (B) Methods.
[0069] Compositions
[0070] The invention provides antimicrobial compositions comprising
one or more acid and one or more organic diol. In one embodiment,
the invention's compositions have an acidic pH. The compositions
may optionally further contain one or more oxidizing agent
(including stabilized oxidizing agent and/or unstabilized oxidizing
agent), and/or one or more surfactant. In particular embodiments,
the acid lacks one or both of --NH group and --NH2 group.
[0071] The invention's compositions are exemplified by, but not
limited to, compositions AA, BB, CC, DD, A, B, C, D, E, F, G, H, I,
J, K, L, M, N and O in Table 1, which are illustrated by
compositions xx1, xx2, xx3, xx4, x1a, x1, x2, x3, x4, x5, x6, x7,
x8, x9, x10, x11, x12, x13, x14 and x15 in Table 2 and by
compositions a1 to a5 in Tables 5 and 8.
TABLE-US-00001 TABLE 1 Exemplary compositions of the
invention.sup.(1) Acid Diol Oxidizing Agent Surfactant Ref..sup.(2)
(wgt. %) (wgt. %) (wgt. %) (wgt. %) AA 0.001-0.9 0.001-2.0
ortho-Phosphoric Propylene acid glycol BB.sup.(7) 0.001-1.0
0.001-2.0 ortho-Phosphoric 1,3-Butanediol acid CC.sup.(8) 0.003-5.0
0.001-2.5 Hydrochloric acid Propylene glycol DD.sup.(7,8) 0.003-5.0
0.001-2.0 Hydrochloric acid 1,3-Butanediol A.sup.(3) 0.001-0.9
0.001-2.0 0.005-2.9 ortho-Phosphoric Propylene Hypochlorous acid
glycol acid B 0.001-0.9 0.001-3.0 0.005-2.9 ortho-Phosphoric
Propylene Hypochlorous acid glycol acid and 0.05-0.9 Bromine
C.sup.(4) 20-30 3 20 ortho-Phosphoric Propylene Hydrogen acid
glycol peroxide D.sup.(5) 30 3 20 0.01-1.9 ortho-Phosphoric
Propylene Hydrogen Sodium lauryl acid glycol peroxide sulfate E
0.001-0.9 0.01-3.0 0.05-0.9 0.01-1.9 ortho-Phosphoric Propylene
Hypochlorous Sodium lauryl acid glycol acid sulfate and 0.05-0.9
Bromine F 0.001-0.05 0.01-3.0 0.05-3.9 ortho-Phosphoric Propylene
Peroxyacetic acid acid glycol G 0.001-0.05 0.01-3.0 0.05-3.9
0.01-1.9 ortho-Phosphoric Propylene Peroxyacetic acid Octyl phenol
acid glycol ethoxylate H 0.001-0.05 0.01-3.0 0.0003-1.9
ortho-Phosphoric Propylene Ozone acid glycol I 0.001-0.05 0.01-3.0
0.0003-1.9 0.01-1.9 ortho-Phosphoric Propylene Ozone Octyl phenol
acid glycol ethoxylate J 1-6 0.1-0.6 0.7-4 ortho-Phosphoric
Propylene Hydrogen acid glycol peroxide K.sup.(6) 30 3 20 Less than
0.1 wgt. % ortho-Phosphoric Propylene Hydrogen of a acid glycol
peroxide surfactant to the concentrate when it is to be applied to
surfaces L 0.3-5.0 0.1-0.6 0.7-4 Hydrochloric acid Propylene
Hydrogen glycol peroxide M.sup.(6) 20 3 20 With or without
Hydrochloric acid Propylene Hydrogen 0.1 wgt. % of a glycol
peroxide surfactant N 0.9-8 0.1-0.6 0.7-4 Citric acid Propylene
Hydrogen glycol peroxide O.sup.(6) 35 3 20 Addition of less Citric
acid Propylene Hydrogen than 0.1 wgt. % of a glycol peroxide
surfactant to the concentrate when it is to be applied to surfaces.
.sup.(1)Water was used as solvent. .sup.(2)pH of compositions other
than concentrated compositions, is acidic. .sup.(3)Concentrated
compositions, such as compositions C, D, K, M, and O are diluted to
a final concentration having an acidic pH less than pH 7.0.
.sup.(4)Composition A is food-safe and does not need to be rinsed
off. .sup.(5)Composition C is a concentrated antimicrobial
composition that has antimicrobial effects when diluted to about
one hundred to one in potable water, is food-safe and does not need
to be rinsed off. .sup.(6)Composition D is a concentrated
antimicrobial composition that has antimicrobial effects when
diluted to about one hundred to one in potable water, is food-safe
and does not need to be rinsed off. .sup.(7)Compositions K, M and O
are concentrated antimicrobial compositions. .sup.(8)With higher
concentrations of 1,3-butanediol, the treated material had a slight
odor. (9) Compositions CC and DD killed Lactobacillus, and broad
leaf material bleached quicker than with compositions AA and
BB.
[0072] The exemplary compositions in Table 1 above are illustrated
by compositions xx1, xx2, xx3, xx4, x1a, x1, x2, x3, x4, x5, x6,
x7, x8, x9, x10, x11, x12, x13, x14 and x15 in Table 2 below (See
also Tables 5 and 8).
TABLE-US-00002 TABLE 2 Additional Exemplary compositions of the
invention. Oxidizing Sub Acid Diol Agent Surfactant Ref..sup.(1)
Ref..sup.(2) (wgt. %) (wgt. %) (wgt. %) (wgt. %) pH.sup.(9) AA xx1
0.035 0.02 ortho- Propylene Phosphoric acid glycol BB xx2 0.03
0.012 ortho- 1,3- Phosphoric acid Butanediol CC xx3 0.01 0.018
Hydrochloric Propylene acid glycol DD xx4 0.008 0.005 Hydrochloric
1,3- acid Butanediol A x1a 0.03 0.02 0.005 4 ortho- Propylene
Hypochlorous Phosphoric acid glycol acid A x1 0.08 0.09 0.05 3-4.1
ortho- Propylene Hypochlorous Phosphoric acid glycol acid B x2 0.08
0.09 0.01 4-5.5 ortho- Propylene Hypochlorous Phosphoric acid
glycol acid and 0.05 Bromine C.sup.(3) x3 21 3 20 0.5-1 ortho-
Propylene Hydrogen Phosphoric acid glycol peroxide D.sup.(4) x4 30
3 20 0.01 0.5-1.5 ortho- Propylene Hydrogen Sodium lauryl
Phosphoric acid glycol peroxide sulfate E.sup.(5) x5 0.07 0.1 0.007
0.03 3.3-4.5 ortho- Propylene Hypochlorous Sodium lauryl Phosphoric
acid glycol acid sulfate and 0.08 Bromine F x6 0.003 0.1 0.5 3-4
ortho- Propylene Peroxyacetic Phosphoric acid glycol acid G x7
0.003 0.14 0.5 0.015 3-4 ortho- Propylene Peroxyacetic octyl phenol
Phosphoric acid glycol acid ethoxylate H x8 0.02 0.1 0.001 5-6
ortho- Propylene Ozone Phosphoric acid glycol I x9 0.02 0.12 0.001
0.015 5.5-6.5 ortho- Propylene Ozone octyl phenol Phosphoric acid
glycol ethoxylate J x10 1 0.1 2.1 less ortho- Propylene Hydrogen
than Phosphoric acid glycol peroxide 2.0 K.sup.(6) x11 30 3 20
0.001 -- ortho- Propylene Hydrogen Cetylpyridinium Phosphoric acid
glycol peroxide chloride L x12 0.3 0.1 3 less Hydrochloric
Propylene Hydrogen than acid glycol peroxide 2.0 M.sup.(7) x13 20 3
20 -- Hydrochloric Propylene Hydrogen acid glycol peroxide N x14 1
0.3 2 less Citric acid Propylene Hydrogen than glycol peroxide 2.0
O.sup.(8) x15 35 3 20 0.001 -- Citric acid Propylene Hydrogen
sodium lauryl glycol peroxide sulfate .sup.(1)Composition reference
letter corresponds to the same reference letter in Table 1.
.sup.(2)Composition sub-reference lettering (e.g., x1) refers to an
exemplary composition within the range of concentrations of the
reference composition (e.g., composition A). .sup.(3)Composition
C-x3 is a concentrated antimicrobial composition.
.sup.(4)Composition D-x4 is a concentrated antimicrobial
composition, and is preferably prepared by first mixing sodium
lauryl sulfate with propylene glycol. .sup.(5)Composition F-x5 is
preferably prepared by first mixing sodium lauryl sulfate with
propylene glycol. .sup.(6)Composition K-x11 is a concentrated
antimicrobial composition. .sup.(7)Composition M-x13 is a
concentrated antimicrobial composition. .sup.(8)Composition O-x15
is a concentrated antimicrobial composition. .sup.(9)pH at
34.degree. F. Concentrated compositions are diluted to a working
concentration having an acidic pH less than 7.0.
[0073] The antimicrobial activity of the exemplary compositions x1
to x15 is shown in Table 6 on bacterial, bacteriophage, viral,
fungal and nematode microbes, using a wide variety of agricultural
products (including leafy vegetables, fruiting and flowering
vegetables, podded vegetables, bulb and stem vegetables, root and
tuberous vegetables, Rosaceae family fruits, Bramble fruits,
berries, Asian fruit, North America fruit, raw meat, raw poultry,
raw seafood, fresh seeds and nuts, eggs, fresh herbs, dried herbs,
animal hides, feathers) and inanimate objects (such as those
encountered in hospitals, food processing plants, residential
buildings, office buildings, etc.). In addition, the antimicrobial
activity of the exemplary compositions al to a5 is shown in Tables
5 and 8 on the highly pathogenic E. coli.
[0074] The components of the invention's compositions include those
that are approved and not approved by the Food and Drug
Administration (FDA) for one or more uses, including, but not
limited to, compounds that are generally recognized as safe (GRAS)
and/or food grade compounds (FCC) and/or registered with the
Environmental Protection Agency (EPA).
[0075] The terms "generally recognized as safe" and "GRAS" when in
reference to a compound refer to each of the substances in 21
C.F.R., .sctn.182 and .sctn.184, approved before 1958 by the FDA
and/or USDA, items published in the Federal Register, as well as
other compounds (even if not on the GRAS list) that are accepted in
the industry as safe for one or more uses by an animal, including
by a human. As used herein, "food grade" and "FCC" refer to the
Food Chemical Codex (FCC) that serves as a guide for food grade
quality consistent with good manufacturing practices and is
published by U.S. Pharmacopeia (USP). For example, and without
limitation, organic diols approved by the FDA and within the scope
of the invention include propylene glycol (1,2-Propanediol),
1,3-butanediol, 1,2-ocatnediol, 1,5-pentanediol, and polyethylene
glycol. Organic diols within the scope of the invention that are
not yet approved by the FDA include ethylene glycol and
1,7-heptanediol.
[0076] The invention's compositions may contain one or more acid.
An "acid" is a chemical compound that, when dissolved in water,
gives a solution with a hydrogen ion activity greater than in pure
water, i.e. has an acidic pH. Acids may be organic or inorganic.
Organic acids are acids that contain carbon. Inorganic acids, also
referred to as mineral acids, are acids that do not contain carbon.
The Chemical Abstracts Service ("CAS"), Columbus, Ohio, USA,
provides a comprehensive repository for data on organic and
inorganic acids. Organic acids include, but are not limited to,
hydroxyethanoic acid (CAS 79-14-1), citric acid (CAS 5949-29-1),
ascorbic acid (CAS 50-81-7), lactic acid (CAS 50-21-5), malic acid
(CAS 617-48-1), octenic acid (CAS 1871-67-6), oxalic acid (CAS
144-62-7), ursolic acid (CAS 77-52-1), and salts thereof. Inorganic
acids include, but are not limited to, ortho-phosphoric acid (CAS
7664-38-2), chromic acid (CAS 1308-38-9), hydrobromic acid (CAS
10035-10-6), hydrochloric acid (also known as muratic acid) (CAS
7647-01-0), nitric acid (CAS 7697-37-2), sulfuric acid (CAS
7664-93-9) and salts thereof. Other acids contemplated within the
scope of the invention include, without limitation, acetic acid,
adipic acid, benzoic acid, glutaric acid, isoascorbic acid,
mandelic acid, propionic acid, salicylic acid, sorbic acid,
succinic acid, tartaric acid (Koefod U.S. Pat. No. 7,090,882),
sodium acid pyrophosphate, acidic sodium hexametaphosphate,
ethylenediaminetetraacetic acid and salts thereof (Andrews et al.,
U.S. Pat. No. 5,490,992). Because of their low solubility in water,
octenic acid and ursolic acid may first be dissolved in an organic
solvent (e.g., glycol, diol, ethanol at a temperature equal to or
greater than 100.degree. F., etc.) before mixing with water. Nitric
acid at some concentrations may be toxic to humans, which makes its
use on foodstuffs less desirable than other acids. Oxalic acid and
chromic acid at some concentrations react with some materials, such
as steel, so their use in cleaning equipment may be less desirable
than other acids. In one embodiment, the acid lacks --NH groups
and/or --NH.sub.2 groups (e.g., is not a sulfamic acid).
[0077] In some embodiments, the amount of acid in the invention's
concentrated compositions may be from 2 to 50 wgt. %, preferably
from 10 to 35 wgt. %, and more preferably from 20 to 30 wgt. %. In
particular embodiments, the concentrate is typically diluted with
water to provide an acid concentration of from 0.001 to 2 wgt. %,
preferably from 0.003 to 1 wgt. %, and more preferably from 0.01 to
0.10 wgt. %.
[0078] The invention' s compositions may contain one or more
organic diol. A "diol" or "glycol" is a chemical compound
containing two hydroxyl (--OH) groups. A diol may be a vicinal diol
or a geminal diol, including an aliphatic diol. A vicinal diol has
hydroxyl groups bonded to adjacent atoms, e.g., ethylene glycol and
propylene glycol. A geminal diol has hydroxyl groups bonded to the
same atom, e.g., methanediol (H.sub.2C(OH).sub.2), 1,3-butanediol,
bisphenol A, etc. Organic diols may contain a branched chain or an
unbranched chain of carbon atoms. Organic diols are preferably
aliphatic diols (i.e., an organic diol composed of carbon and
hydrogn and does not contain an aromatic ring), and more preferably
simple aliphatic diols. A "simple aliphatic diol" is an aliphatic
diol in which all the carbon atoms are covalently bonded to only
hydrogen, and is exemplified by ethylene glycol, propylene glycol,
1,3-butanediol, etc. In contrast, a "complex aliphatic diol" is an
aliphatic diol in which at least one carbon atom is covalently
bonded to one or more atoms other than hydrogen. Thus, a complex
aliphatic diol may contain --COOH, --NH.sub.2, etc. In some
embodiments, organic diols may contain one or more ether group
(glycol ether) and/or one or more ester group (glycol ester),
including monoesters and diesters. In another embodiment, organic
diols may have one hydroxyl group esterified with a saturated
and/or unsaturated aliphatic acid other than benzoic acid, and the
other hydroxyl group esterified with the same or different
saturated and/or unsaturated aliphatic acid other than benzoic
acid. Exemplary organic diols include, without limitation,
propylene glycol (CAS 57-55-6), ethylene glycol (CAS 107-21-1),
1,3-butanediol (CAS 107-88-0), 1,7-heptanediol (CAS 629-30-1),
1,2-octanediol (CAS 1117-86-8), 1,5-pentanediol (CAS 111-29-5) and
polyethylene glycol (CAS 25322-68-3). In some embodiments, organic
diols contain from 2 to 20, such as from 2 to 10, and/or such as
from 2 to 5 carbon atoms. For example, the diol ethylene glycol
contains 2 carbon atoms, 1,3-butanediol contains 4 carbon atoms,
1,5-pentanediol contains 5 carbon atoms, 1,7-heptanediol contains 7
carbon atoms, and 1,2-octanediol contains 8 carbon atoms.
[0079] In some embodiments, the amount of organic diol in the
invention's concentrated compositions may be from 2 to 5 wgt. %,
preferably from 3 to 4 wgt. %, and more preferably 3 wgt. %. In
particular embodiments, the concentrate is typically diluted with
water to provide an organic diol concentration of from 0.01 to 1
wgt. %, preferably from 0.09 to 0.3 wgt. %, and more preferably
from 0.09 to 0.15 wgt. %.
[0080] The invention's compositions may contain one or more
oxidizing agent. "Oxidizing agent," "oxidant" and "oxidizer" are
interchangeably used to refer to a chemical compound that transfers
oxygen atoms or gains electrons in a redox chemical reaction. In
both cases, the oxidizing agent becomes reduced in the process.
Exemplary oxidizing agents include, without limitation, bromine
(CAS 7726-95-6), hypochlorous acid (CAS 7790-92-3, also referred to
as IUPAC name chloric(I) acid), hydrogen peroxide (CAS 7722-84-1),
peroxyacetic acid (PAA) (CAS 79-21-0), and ozone (CAS 1-28-15-6).
Hypochlorous acid cannot be isolated in pure form, but can be
derived from, without limitation, sodium hyporchlorite, calcium
hyporchlorite, chlorine dioxide and chlorine. Further oxidizing
agents include sodium bromide, peracetic acid, chlorine dioxide,
sodium hypochlorite, hydrogen peroxide (Bautista U.S. Pat. No.
6,045,846) and ozone. Ozone may be introduced into a fluid using
methods known in the art (Conners et al., U.S. Pat. No.
6,086,833).
[0081] In preferred embodiments, the invention's compositions
contain more than one oxidizer to produce a greater reduction in
antimicrobial activity and/or in exposure times when compared to
single oxidizers in order to obtain the same antimicrobial
activity.
[0082] In some embodiments, the amount of oxidizer in the
invention's concentrated compositions may be from 2 to 30 wgt. %,
preferably from 10 to 30 wgt. %, and more preferably from 10 to 20
wgt. %. In particular embodiments, the concentrate is typically
diluted with water to provide an oxidizer concentration of from
0.01 to 5 wgt. %, preferably from 0.001 to 3 wgt. %, and more
preferably from 0.01 to 1 wgt. %. For example, levels of
hypochlorous acid and hypochlorite are reported as C1.sub.2, using
the standard DPD (N,N-diethyl-p-phenylenediamine) colorimetric test
method (Hach Chemical Co., Loveland Colo.), described in Harp, U.S.
Pat. No. 5,362,650 and Kroll, U.S. Pat. No. 6,180,412, by employing
standard instruments, such as model number 5870000 pocket
colorimeter II "2."
[0083] In contrast to the art (e.g., Williams, U.S. Patent
Application 20060003023), in particular embodiments, the
invention's compositions contain an "unstabilized oxidizer"
("USOX"), i.e., contain an oxidizing agent in the absence of an
oxidizing agent stabilizer. A "compound that stabilizes an
oxidizing agent" and "oxidizing agent stabilizer" interchangeably
refer to a compound that reduces the rate of decrease in
concentration of an oxidizing agent, such as that resulting from
degradation of the oxidizer. Oxidizing agent stabilizers include,
for example, sulfamic acid, alkali metal sulfamate (Williams, U.S.
Patent Application 20060003023), amide derivatives of carbonic
acid, hydrogen cyanide, carboxylic acid, amino acid, sulfuric acid,
phosphoric acid, boric acid, urea, thiourea, creatinines, cyanuric
acids, alkyl hydantoins, mono-ethanolamine, di-ethanolamine,
organic sulfonamides, biuret, sulfamic acid and salts thereof,
organic sulfamic acid, melamine (Shim et al., U.S. Pat. Nos.
6,478,972 and 6,533,958).
[0084] Oxidizers are unstable, and their rate of oxidation may
change (i.e., increase or decrease) and may become uncontrolled,
resulting in depletion of the oxidizing agent in a short period of
time (such as less than 6 hours, Example 9). To avoid this problem,
the prior art, such as Self et al., U.S. Pat. No. 3,328,294,
discloses a stabilized oxidizer ("SOX") produced by reacting the
unstabilized oxidizer NaOBr with an equal molar ratio of the
stabilizer sulfamic acid. They disclose that the reaction does not
go un-controlled and the stabilized oxidizer may linger for days.
Because of these characteristics they caution against the use of
their compositions on foods and in potable water.
[0085] In contrast to the prior art, particular embodiments, the
invention uses unstabilized oxidizers in order to take advantage of
newer technology and increase the log microbial kill rate. It is
the inventor's view that unstabilized oxidizers provide an
instantaneous reaction in a process stream or on contacting a
surface, a reduction in toxicity to humans and animals, an almost
immediate kill of contacted microbes, shorter processing time,
instantaneous concentration control at product contact points, and
take advantage of new control systems for fast automatic control of
active ingredients.
[0086] The invention's compositions may contain one or more
surfactant. A "surfactant" is a chemical compound that lowers the
surface tension of a liquid, allowing easier spreading of the
liquid, and thereby acting as a wetting agent. In one embodiment, a
surfactant is an organic compound that is amphiphilic, meaning that
it contains one or more hydrophobic groups ("tails") and one or
more hydrophilic groups ("heads"). Surfactants may be anionic,
neutral or cationic. "Anionic surfactants" are surfactants that
dissolve in water to release an anion, and include, for example,
sodium lauryl ether sulfate, also referred to as sodium lauryl
sulfate (CAS 009004-82-4), ammonium lauryl sulfate (CAS 2235-54-3),
alkylbenzene sulfonic acid (CAS 27176-87-0), sodium 2-ethylhexyl
sulfate (CAS 126-92-1), and dioctyl sodium sulfosuccinate (Andrews
et al., U.S. Pat. No. 5,490,992). "Neutral surfactants" are
surfactants that dissolve in water without releasing an anion or
cation, and include, without limitation, octyl phenol ethoxylate
(CAS 9002-93-1), glyceryl monostearate (CAS 31566-31-1),
polyglyceryl-10 decaoleate (CAS 011094-60-3), and lauryl lactyl
lactate (CAS 910661-93-7). "Cationic surfactants" are surfactants
that dissolve in water to release a cation, and include the
exemplary cetrimonium bromide (CAS 57-09-0), cetylpyridinium
chloride (CAS 123-03-5), benzalkonium chloride (CAS 8001-54-5), and
cocamidopropyl betaine (CAS 86438-79-1). Additional examples of
surfactants include those that contain ethylene oxide moieties
and/or propylene oxide moieties. Yet more examples of surfactants
include linear alkylbenzene sulfonates, alcohol sulfates,
alpha-olefin sulfonates, alcohol ethoxylates, nonylphenyl
ethoxylates, alkylpolyglucosides, fatty alkanoamides, fatty amine
oxides, sodium dioctylsulfosuccinate, dodecylbenzene sulfonic acid
and salts thereof, the sodium salt of sulfonated oleic acid, sodium
dodecylbenzene sulfonate, dodecyidiphenyloxidedisulfonic acid and
salts thereof (Koefod et al., U.S. Pat. No. 7,090,882, Andrews et
al., U.S. Pat. No. 5,490,992).
[0087] In some embodiments, the amount of surfactant in the
invention's concentrated compositions may be from 0.05 to 0.5 wgt.
%, preferably from 0.01 to 0.03 wgt. %, and more preferably from
0.01 to 0.15 wgt. %. In particular embodiments, the concentrate is
typically diluted with water to provide a surfactant concentration
of from 0.01 to 0.05 wgt. %, preferably from 0.001 to 0.005 wgt. %,
and more preferably from 0.001 wgt. % to 0.03 wgt. %.
[0088] The invention additionally provides compositions comprising
a concentrated solution of any of the compositions disclosed
herein. A "concentrate" of a first composition refers to a
compositions that contains the same components and the same ratio
of these components (relative to each other) as in the first
composition, wherein the amount of each component in the first
composition is multiplied by the same fold factor to yield the
amount of that component in the concentrated composition. The fold
factor may be from 2 to 100,000, from 5 to 10,000, from 10 to
1,000, and from 100 to 500. For example, for a composition
containing 4 wgt. % component A and 6 wgt. % component B, a 2-fold
concentrate of this composition contains 8 wgt. % component A and
12 wgt. % component B, and a 3-fold concentrate of this composition
contains 12 wgt. % component A and 18 wgt. % component B.
Concentrates may be used directly or diluted. They provide ease of
handling and shipping (because of smaller volumes). Exemplary
concentrates are compositions C, D, J, K, M, O in Table 1, as
exemplified by compositions x3, x10, x11, x13 and x15 of Table
2.
[0089] The invention also contemplates compositions comprising
diluted solutions of any of the compositions disclosed herein. A
"diluted" solution of a first composition refers to a compositions
that contains the same components and the same ratio of these
components (relative to each other) as in the first composition,
wherein the amount of each component in the first composition is
divided by the same fold factor to yield the amount of that
component in the concentrated composition. The fold factor may be
from 2 to 100,000, from 5 to 10,000, from 10 to 1,000, and from 100
to 500. For example, for a composition containing 4 wgt. %
component A and 6 wgt. % component B, a 2-fold dilution of this
composition contains 2 wgt. % component A and 3 wgt. % component B.
Diluted solutions may be used directly or further diluted.
Exemplary diluted solutions are compositions A, B,E, F, G, H, I, L,
N, O in Table 1, as exemplified by compositions xx1, xx2, xx3, xx4,
x1a, x1, x2, x4, x5, x6, x7, x8, x9, x12, x14 and in Table 2 and by
compositions a1 to a5 of Tables 5 and 8.
[0090] The invention's compositions may be prepared by adding and
mixing the acid alone into water, followed by adding and mixing in
the diol, and then adding and mixing in the oxidizing agent.
Surfactant may be subsequently added and mixed in. The compositions
may be compounded at temperatures between the freezing point and
the boiling point of the solvent (e.g., water) used.
[0091] When making concentrates, the diol and acid may be mixed
together in concentrated quantities, followed by the surfactant, if
desired, for delivery to a mixture of oxidizer and solvent (e.g.,
water). Alternatively, the diol and oxidizer may be mixed together
in concentrated quantities for delivery to a mixture of acid and
solvent (e.g., water). Concentrates may be stored for more than 1
week.
[0092] The invention's compositions may be used in a variety of
ways as further described below.
[0093] B. Methods
[0094] In one embodiment, the invention's compositions have
antimicrobial activity and may be used in methods for reducing the
number of microbes on a surface, comprising (a) providing (i) a
first surface comprising microbes, (ii) an antimicrobially
effective amount of any of the invention's compositions, and b)
contacting the surface with the composition under conditions that
produce a contacted surface comprising a reduced number of the
microbes compared to the number of the microbes on the first
surface in the absence of the contacting step.
[0095] The term "antimicrobial" and "antimicrobial activity" when
in reference to a compound refers to a compound that reduces the
number of and/or rate of growth of a microbe compared to the number
and/or rate of growth of the microbe in the absence of the
compound. In one embodiment, the number of and/or rate of growth of
a microbe in the presence of an antimicrobial compound is at least
10% lower than, at least 25% lower than, at least 50% lower than,
at least 75% lower than, at least 90% lower than, at least 99%,
and/or at least 100% lower than the number of and/or rate of growth
of the microbe in the absence of the antimicrobial compound. In
another embodiment, a 9-log number of microbe is reduced by at
least 3-log, at least 5-log, at least 7-log, and/or at least 9-log
in the presence of the antimicrobial compound.
[0096] An "antimicrobially effective amount" of a compound or
composition refers to an amount of the compound or composition that
has antimicrobial activity, including microbistatic amounts and
microbicidal amounts.
[0097] "MIC" and "minimum inhibitory concentration" refer to the
lowest concentration that is microbistatic under a specific set of
conditions.
[0098] "MBC" and "minimum biocidal concentration" refer to the
lowest concentration that is microbicidal under a specific set of
conditions.
[0099] An antimicrobial can be antibacterial, antifungal, antiviral
and/or antinematode. An antimicrobial can be microbistatic,
microbicidal, or both. An antimicrobial is "microbistatic" (e.g.,
bacteriostatic, fungistatic, etc.) if it reduces cell division by
an amount less than 100%, without or without reducing cell
viability. An antimicrobial is "microbicidal" (e.g., bactericidal,
fungicidal, etc.) if it reduces cell viability by 100%, i.e.,
causes 100% cell death. Cell death is commonly detected by the
absence of cell growth in liquid growth medium (e.g., absence of
turbidity) or on a solid surface (e.g., absence of colony formation
on agar). A "sterilizer," "sanitizer" and "disinfectant" are
microbicidal. In contrast, a "preservative" is microbistatic.
Certain microbistatic compositions are not bactericidal at any
concentration.
[0100] Those of skill in the art know that a composition may have
microbistatic or microbicidal activity by altering the
concentration of its components, temperature, and contact time with
a surface. For example, a composition that is microbistatic at a
given concentration may be microbicidal at a higher concentration.
Also, a composition that is microbistatic at a given temperature
may be microbicidal at a higher temperature. Similarly, a
composition that is microbistatic at a particular contact time may
be microbicidal at a longer contact time with a surface.
[0101] Methods for determining antimicrobial activity are known in
the art, e.g., Andrews et al., U.S. Pat. No. 5,490,992, Bailey et
al., U.S. Pat. No. 4,107,192and disclosed herein. For example, one
assay involves exposing a bacterial strain (e.g., Escherichia coli)
to a test composition on a particular substrate (e.g., an
agricultural product) at a predetermined bacterial level in a
culture media at an appropriate temperature. After a sufficient
amount of contact time, an aliquot of a sample containing the
exposed bacteria is collected, diluted and plated on agar. The
plated sample of bacteria is then incubated for about forty eight
(48) hours and the number of viable bacterial colonies growing on
agar is counted. Once colonies have been counted the reduction in
the number of bacteria caused by the test composition is
determined. Reduction in bacteria is typically reported as the
difference between the log.sub.10 of the initial inoculant's count
and the log.sub.10 of the inoculant's count after exposure to the
test composition. Assays may also be subjective, such as by
visually observing a difference in the amount of spoilage of
agricultural products.
[0102] The invention's compositions exhibit surprising synergistic
activity with respect to, for example, antimicrobial activity of
their components. "Synergistic" activity of a combination of two or
more components means that the activity of the combination of two
or more components is greater than the activity of each component
alone. Without intending to limit the invention to any particular
method of calculation, in one embodiment, synergy may be determined
using the industrially accepted method described by Kull et al.,
"Mixtures of Quaternary Ammonium Compounds and Long-chain Fatty
Acids as Antifungal Agents," Applied Microbiology 9:538-541 (Apr.
17, 1961) (see also Williams, U.S. Patent Application 20060003023),
using the ratio determined by the formula:
Q.sub.a/Q.sub.A+Q.sub.b/Q.sub.B=Synergy Index ("SI") wherein:
[0103] Q.sub.A=concentration of compound A (first component) in
ppm, acting alone, which produced an end point (MIC or MBC of
Compound A).
[0104] Q.sub.a=concentration of compound A in ppm, in the mixture,
which produced an end point.
[0105] Q.sub.B=concentration of compound B (second component) in
ppm, acting alone, which produced an end point (MIC or MBC of
Compound B).
[0106] Q.sub.b=concentration of compound B in ppm, in the mixture,
which produced an end point.
[0107] When the sum of Q.sub.a/Q.sub.A and Q.sub.b/Q.sub.B is
greater than one, antagonism is indicated. When the sum is equal to
one, additivity is indicated, and when less than one, synergism is
demonstrated. The lower the SI, the greater the synergy shown by
that particular mixture.
[0108] Thus, in one embodiment, synergy of the combination of acid
and organic diol is obtained where the amount of each of the acid
alone and of the organic diol alone has lower antimicrobial
activity compared to the antimicrobial activity of a combination of
the acid and the organic diol. In another embodiment, synergy of
the combination of acid, organic diol and oxidizing agent is
obtained where the amount of each of (a) the acid alone, (b) the
organic diol alone, (c) the oxidizing agent alone, (d) combination
of the acid and the organic diol, (e) combination of the acid and
the oxidizing agent, and (f) combination of the organic diol and
the oxidizing agent, has lower antimicrobial activity compared to
the antimicrobial activity of a combination of the acid, the
organic diol and the oxidizing agent.
[0109] For example, Table 3A shows antimicrobial synergy of
propylene glycol and phosphoric acid, synergy of propylene glycol,
phosphoric acid and hydrogen peroxide, and synergy of propylene
glycol, phosphoric acid, hydrogen peroxide and sodium lauryl
sulfate.
[0110] Also, Table 4 shows antimicrobial synergy of two components
when compared to single components of Table 3A.
[0111] In another example, data herein shows synergistic
antimicrobial activity of the components of composition "a1" that
contains hypochlorous acid 0.01 wgt. %, phosphoric acid 1 wgt. %,
and propylene glycol 2 wgt. %.
[0112] In particular, Table 3B shows that 180 minute treatment of
spinach with each of hypochlorous acid 0.01 wgt. %, phosphoric acid
1 wgt. % and propylene glycol 2 wgt. % alone in water causes a
5.01-log, 4.13-log, and 3.98-log reduction, respectively, in a
6.87-log E. coli inoculum. Table 4 shows similarly low
antimicrobial activity by a combination of two components of
hypochlorous acid 0.01 wgt. % and phosphoric acid 1 wgt. %, which
resulted in a 4.9-log reduction in a 6.87-log E. coli inoculum. In
contrast, Table 5 shows that 180 minute treatment of spinach with
the combination "a1" that contained hypochlorous acid 0.01wgt. %,
phosphoric acid 1 wgt. %, and propylene glycol 2 wgt. % resulted in
a 6.57-log reduction in a 6.57-log E. coli inoculum.
[0113] Similarly, Table 3B shows that 180 minute treatment of
spinach with each of bromine 0.08 wgt. %, phosphoric acid 1 wgt. %
and propylene glycol 2 wgt. % alone caused a 4.44-log, 4.13-log,
and 3.98-log reduction, respectively, in a 6.87-log E. coli
inoculum. Table 4 shows similarly low antimicrobial activity by a
combination of two components of bromine 0.08 wgt. % and phosphoric
acid 1 wgt. % which resulted in a 4.88-log reduction in a 6.87-log
E. coli inoculum. In contrast, Table 5 shows that 180 minute
treatment of spinach with the combination "a2" that contained
bromine 0.08 wgt. %, phosphoric acid 1 wgt. % and propylene glycol
2 wgt. % resulted in a 6.49-log reduction in a 6.57-log E. coli
inoculum.
[0114] In yet another example, Table 3B shows that 180 minute
treatment of spinach with each of hypochlorous acid 0.01 wgt. %,
bromine 0.05 wgt. %, phosphoric acid 1 wgt. %, and propylene glycol
2 wgt. % alone in water caused a 5.01-log, 4.44-log, 4.13-log and
3.98-log reduction, respectively, in a 6.87-log E. coli inoculum.
Table 4 shows similarly low antimicrobial activity by a combination
of two components of bromine 0.08 wgt. % and phosphoric acid 1 wgt.
% which resulted in a 4.88-log reduction in a 6.87-log E. coli
inoculum. In contrast, Table 5 shows that 180 minute treatment of
spinach with the combination "a5" that contained hypochlorous acid
0.01 wgt. %, bromine 0.05 wgt. % , phosphoric acid 1 wgt. %, and
propylene glycol 2 wgt. %, resulted in a 6.57-log reduction in a
6.57-log E. coli inoculum.
[0115] The invention's compositions showed very effective
antimicrobial activity (Tables 5, 6, 8 and 9). For example, Table 6
shows the antimicrobial activity of the invention's compositions on
bacterial, bacteriophage, viral, fungal and nematode microbes,
using a wide variety of agricultural products (including leafy
vegetables, fruiting and flowering vegetables, podded vegetables,
bulb and stem vegetables, root and tuberous vegetables, Rosaceae
family fruits, Bramble fruits, berries, Asian fruit, North America
fruit, raw meat, raw poultry, raw seafood, fresh seeds, edible
sprouts, and nuts, eggs, fresh herbs, dried herbs, spices, animal
hides, feathers) and inanimate objects (such as those encountered
in hospitals, food processing plants, residential buildings, office
buildings, etc.). Tables 3-5 and 6 also show that the invention's
compositions produced at least a 3-log reduction in microbial
inoculums that contained microbes ranging from 3-log to 8.78-log,
after treatment for 2 minutes or longer at a temperature of at
least 34.degree. F. In addition, Tables 5 and 8 show 100% reduction
in E. coli on spinach and strawberry after 180-minute treatment at
about 33.degree. F. with several of the invention's compositions.
Additional exemplary compositions of the invention in Table 6 show
100% reduction in microbes when treating inanimate objects.
[0116] One surprising property of the invention's compositions is
that they retain their antimicrobial activity over a broad range of
temperatures, including from the freezing temperature of the
composition to the temperature of autoclaving (about 120.degree.
C.). Thus, the invention's compositions may be used as
antimicrobials at temperatures from the freezing temperature of the
composition to 20.degree. C., to 34.degree. C., to 40.degree. C.,
to 60.degree. C., to 80.degree. C., to 100.degree. C. and/or to
120.degree. C. For example, Table 6 shows that contacting shrimp,
that was contaminated with E. coli, for 30 minutes with the
invention's compositions that had previously been frozen into ice
cubes (about 29.degree. F.) resulted in a 6.5-log reduction in E.
coli from 6.87-log.
[0117] Thus, the invention's compositions advantageously retain
their antimicrobial (and other) activities at temperatures equal to
or below room temperature. "Room temperature" and "ambient
temperature" are used interchangeably to refer to 21 degrees
celcius (.degree. C.), equivalent to 70 degrees Fahrneheit
(.degree. F.). "Chilling temperature" refers to a temperature less
than room temperature, i.e., less than 21.degree. C. (equivalent to
less than 70.degree. F.), including, but are not limited to,
temperatures from 30.degree. F. to 69.degree. F., from 34.degree.
F. to 45.degree. F., from 34.degree. F. to 69.degree. F., from
40.degree. F. to 42.degree. F., 40.degree. F. to 69.degree. F. and
from 50.degree. F. to 69.degree. F. Temperatures greater than room
temperature include, but are not limited to, temperatures from
71.degree. F. to 220.degree. F., from 100.degree. F. to 220.degree.
F. and from 150.degree. F. to 220.degree. F. Thus, the invention's
compositions are particularly useful at 30.degree. F. to 40.degree.
F., preferably at 34.degree. F., which are the temperatures at
which many food products are preferably maintained prior to human
consumption, such as in the field, during transport from the field,
and before, during and/or after processing for human consumption.
For agricultural products that may be ingested raw, in several
embodiments, compositions brought in contact with these products
are preferably formulated to contain one or more GRAS listed
component.
[0118] Thus, the invention's compositions may be applied to fresh
produce and ornamental plants at refrigerated temperatures of about
34.degree. F. In another embodiment, the invention's compositions
may be used when frozen, such as ice cubes or crushed ice. The
temperature at which the compositions of the invention freeze will
vary depending on the type and concentration of components in the
composition. The invention's compositions are also useful
antimicrobials at room temperature. For example in strawberries and
tomatoes, a reduction in slime and mold was observed over a
two-week period of storage at 75.degree. F. following washing with
the invention's compositions.
[0119] The invention's compositions are also useful for reducing
odor associated with microbial breakdown of organic material, e.g.,
food.
[0120] In addition, the invention's compositions surprisingly
reduced discoloration (e.g., pinking and browning) of agricultural
products. Thus, in addition to the compositions' preservative
effects during a two-week storage at 40.degree. F. on varieties of
lettuce and cabbage, a reduction in pinking and browning was also
observed when applying anti-discoloration effective amounts of the
invention's compositions. An "anti-discoloration effective amount"
of a compound refers to an amount of the compound that reduces the
level of discoloration compared to the level in the absence of the
compound.
[0121] "Discoloration" refers to a change in color and/or hue. For
example, discoloration of agricultural products includes pinking
and browning that are generally observed after cutting produce
and/or during storage. "Pinking" refers to a de novo development
and/or increase in pink, red and/or rust color and/or hue such as
that observed after storage at the cut edges of lettuce, especially
iceberg lettuce, cabbage, celery, and Bok Choy. "Browning" refers
to de novo development and/or increase in brown, grey and/or green
color and/or hue such as that observed after storage at the cut
edges of lettuce, cabbage, celery, Bok Choy, potatoes, parsnips,
avocado, apples, strawberries, spinach, as well as after storage of
meat (such as hamburger), poultry and seafood.
[0122] The level of discoloration may be subjectively determined
using known methods (Schwank, U.S. Pat. No. 5,087,467) as well as
using blind tests as previously described for other attributes such
as crispness, color, and appearance of freshness (Estrada, U.S.
Pat. No. 5,599,571). For example, at 3, 6 and 14 days after
treatment with the invention's compositions, the agricultural
produce is rated as having excellent, good, fair or poor color in a
blind test by four individuals, and is compared with produce that
is treated under the same conditions with the exception of omission
of the invention's composition.
[0123] The invention's compositions may be applied by routine
methods for application of antimicrobial solutions such as
immersing, dipping, spraying, fogging, aerosoling, washing and
scrubbing. "Fogging" refers to production of liquid droplets of 0.2
.mu.m, or smaller, diameter, e.g., by applying differential
pressure to the liquid. "Aerosoling" refers to production of liquid
droplets of greater than 0.2 .mu.m diameter, e.g., by applying high
pressure to the liquid.
[0124] For large scale processing of produce (e.g., fruits and
vegetables), a suitable volume of a working solution of the
invention's compositions may be contained in a tank or flume,
having agitation. The produce is deposited into this vessel for a
period sufficient to reduce microbial contamination. The treated
produce is then removed and transported down the line for further
processing. Final rinsing is not necessary as the amount of residue
is very low or non-toxic (Table 9), thus processing may include a
step that leaves only a very thin, adherent film on the product's
surfaces. This film furnishes an extended period of antimicrobial
activity without adversely altering the color, texture and/or odor
of the product surfaces during an extended shelf life.
[0125] In addition to the tank or flume, other methods are
available by which the invention's compositions and their
concentrates may be brought to contact the target microorganisms on
any surface, including, without limitation, closed pipes,
low-pressure spray, high-pressure spray, fog, vaporization, sonic
energy, aerosols, wiping, immersion, scrubbing, swabbing, mopping,
brushing, and gels. Contacting the invention's compositions with
target microorganisms on any surface may be accomplished at
atmospheric pressure and/or under a vacuum, i.e., at a pressure
less than atmospheric pressure.
[0126] The invention's compositions may also be used during the
processing of meat and poultry as previously described (Andrews et
al., U.S. Pat. No. 5,490,992; Shane et al., U.S. Patent Application
US 2002/0134317) and further described herein. For example, live
animals or poultry are completely contacted with one or more of the
invention's dilute compositions using best practices method. The
treatment time is determined by whether subsequent processing is
desired with a wet animal or whether it should appear to be dry.
Without rinsing after this application, the invention's composition
exhibits residual activity and substantially reduces the total
microbial load. After the slaughter step, an animal carcass may be
contacted with a more concentrated composition of the invention,
and most preferably one that contains a surfactant, whereas one or
more of the invention's dilute compositions may be used in
subsequent steps according to best practices. The cutting and
trimming workstations ideally should be cleaned and completely
contacted with one or more of the invention's concentrated
compositions between each carcass. In the case of poultry, where
cross contamination is highly possible due to common wash and
chiller tanks, one or more of the invention's dilute compositions
may be used in these tanks. The last drainage step may be performed
without the need for rinsing and the carcasses can be packaged
normally. The antimicrobial composition is still effective at the
low chilling temperatures and very low amounts.
[0127] Seeds and grains used for raw sprouts and/or other ingestion
purposes are normally difficult to treat without harm and
deterioration. However, contacting these products with one or more
of the invention's dilute compositions and drying without the need
for rinsing can substantially improve the product quality and
safety.
[0128] Additional exemplary uses of the invention's methods are
described herein in the Examples and Table 7 for treating fruits
and vegetables, raw eggs, poultry with and without feathers,
poultry feathers, meat, hides, seeds and nuts, seafood, dried
goods, inanimate objects, skin, reverse osmosis membranes, etc.
[0129] After contact with the invention's composition, the treated
products may be rinsed. Alternatively, the treated products may be
allowed to dry (e.g., at ambient or chilling temperatures) or dried
by spinning, heating, toweling, etc., without rinsing. One
advantage of the invention's compositions is that, after drying,
they form a very thin film on the surface of the treated .products.
This film is edible because of its low toxicity to higher life
forms, while continuing to exhibit antimicrobial activity, thus
eliminating the need for rinsing off the invention's composition
from the treated agricultural (or other) products.
[0130] The invention's compositions show antibacterial activity
against a wide spectrum of microbes, including bacterial, viral,
fungal and nematode microbes. For example, data herein (Tables 3-6
and 8) show the antimicrobial activity of the invention's exemplary
compositions on E. Coli (ATCC 25922) and the highly pathogenic E.
coli O157:H7 strain (ATCC 43888) and Salmonella enterica (ATCC
10708).
[0131] The invention's compositions also show antibacterial
activity against Gram-positive bacteria. Data herein (Table 6) show
the antimicrobial activity of the invention's exemplary
compositions on Staphylococcus aureus (ATCC 25923) and to a lesser
extent on Lactobacillus sp. (ATCC 55326).
[0132] In addition, the invention's compositions show antifungal
activity. Data herein (Table 6) show the antimicrobial activity of
the invention's exemplary compositions on the fungi Aspergillus
flavus (ATCC 15517) and Septoria lycopersici (ATCC Q99324) and
toxic mold Stachybotrys chartarum (ATCC 9182).
[0133] The invention's compositions also show antiviral activity.
Data herein (Table 6) show the antimicrobial activity of the
invention's exemplary compositions on rhinovirus (ATCC vr1110).
[0134] Moreover, the invention's compositions show
antibacteriophage activity. Data herein (Table 6) show the
antimicrobial activity of the invention's exemplary compositions on
bacteriophage T4 (ATCC 35060-B4).
[0135] Furthermore, the invention's compositions show antinematode
activity. Data herein (Table 6) show the antimicrobial activity of
the invention's exemplary compositions on Aphelenchoides fragariae
(ATCC 12974), which is a nematode that destroys plant crops,
necessitating burning of crops to eradicate the nematode.
[0136] One of the surprising properties of the invention's
compositions is that they show a differential antimicrobial
activity on pathogenic and non-pathogenic microbes. For example, a
differential antimicrobial effect was observed between
Gram-positive pathogenic and non-pathogenic bacteria. In
particular, Table 6 shows the surprising result that exemplary
composition "x4" of the invention caused greater than a 4-log
reduction in a 4.21-log inoculum of Staphylococcus aureus (ATCC
25923) while the exemplary composition "x3" caused less than 2-log
reduction in a 8.78-log inoculum of Lactobacillus sp (ATCC
55326).
[0137] Also, a differential antimicrobial activity was observed
between pathogenic Gram-negative bacteria and non-pathogenic
Gram-positive bacteria. In particular, Table 6 shows the surprising
result that exemplary compositions x1, x2, x4, x5, x7, x9, x10,
x11, x12, x13 and x15 of the invention caused greater than a 6-log
reduction in about 6.87-log inoculum of E. coli (ATCC 25922 or ATCC
43888) while the exemplary composition "x3" caused less than 2-log
reduction in a 8.78-log inoculum of Lactobacillus sp (ATCC 55326).
Similarly, Table 6 shows the surprising result that exemplary
compositions x4, x7, x10 and x11 of the invention caused greater
than 5-log reduction in a 5.45-log inoculum of S. enterica (ATCC
25922) while the exemplary composition x3 caused less than 2-log
reduction in a 8.78-log inoculum of Lactobacillus sp (ATCC
55326).
[0138] Not only was the differential antimicrobial activity
observed between pathogenic and non-pathogenic bacteria, but it was
also observed between pathogenic bacteria and non-pathogenic
bacteriophage. For example, Table 6 shows the surprising result
that exemplary compositions "x1" of the invention caused greater
than a 6.8-log reduction in a 6.88-log inoculum of E. coli (ATCC
25922) while the same exemplary composition "x1" caused less than
1-log reduction in a 8.78-log inoculum of bacteriophage T4 (ATCC
35060-B4).
[0139] Additional differential antimicrobial activity was observed
between pathogenic and non-pathogenic viruses. For example, Table 6
shows the surprising result that exemplary compositions "x13" of
the invention caused a 2.4-log100% 2.4-log (i.e., 100%) reduction
in a 2.4-log inoculum of pathogenic Rhinovirus (ATCC vr1110) while
composition "x1" caused less than 1-log reduction in a 8.78-log
inoculum of non-pathogenic bacteriophage T4 (ATCC 35060-B4).
[0140] The invention's compositions are useful as antimicrobials in
applications to a varievariety of surfaces, including agricultural
products and inanimate objects.
[0141] In one embodiment the agricultural product is a leafy
vegetable, as exemplified by, but not limited to, Amaranth
(Amaranthus cruentus), Beet greens (Beta vulgaris subsp. vulgaris),
Bitterleaf (Vernonia calvoana), Bok choy (Brassica rapa Chinensis
group), Brussels sprout (Brassica oleracea Gemmifera group),
Cabbage (Brassica oleracea Capitata group), Ceylon spinach (Basella
alba), Chicory (Cichorium intybus), Chrysanthemum leaves
(Chrysanthemum coronarium), Corn salad (Valerianella locusta),
Cress (Lepidium sativum), Dandelion (Taraxacum officinale), Lettuce
(Lactuca sativa), Mizuna greens (Brassica rapa Nipposinica group),
Mustard (Sinapis alba), Napa/Chinese Cabbage (Brassica rapa
Pekinensis group), New Zealand Spinach (Tetragonia tetragonioides),
Orache (Atriplex hortensis), Pea sprouts/leaves (Pisum sativum),
Sea beet (Beta vulgaris subsp. maritima), Seakale (Crambe
maritima), Soko (Celosia argentea), Spinach (Spinacia oleracea),
Swiss chard (Beta vulgaris subsp. cicla var. flavescens), Turnip
greens (Brassica rapa Rapifera group), Watercress (Nasturtium
officinale), Water spinach (Ipomoea aquatica), and Yau choy
(Brassica napus).
[0142] In another embodiment, the agricultural product is a
fruiting and/or flowering vegetable, as exemplified by, but not
limited to, Armenian cucumber (Cucumis melo Flexuosus group),
Eggplant or Aubergine (Solanum melongena), Avocado (Persea
americana), Bell pepper (Capsicum annuum), Bitter melon (Momordica
charantia), Cayenne pepper (Capsicum frutescens), Chayote (Sechium
edule), Chili pepper (Capsicum annuum Longum group), Cucumber
(Cucumis sativus), Globe Artichoke (Cynara scolymus), Perennial
cucumber (Coccinia grandis), Pumpkin (Cucurbita maxima, Cucurbita
pepo), Pattypan squash, Squash (aka marrow) (Cucurbita pepo), Corn
(Zea mays), Sweet pepper (Capsicum annuum Grossum group), Tomato
(Solanum lycopersicum), Tomatillo (Physalis philadelphica), Winter
melon (Benincasa hispida), and Zucchini or Courgette (Cucurbita
pepo).
[0143] In a further embodiment, the agricultural product is a
podded vegetable, as exemplified by, but not limited to, American
groundnut (Apios americana), Azuki bean (Vigna angularis),
Black-eyed pea (Vigna unguiculata subsp. unguiculata), Chickpea
(Cicer arietinum), Fava bean (Vicia faba), Indian pea (Lathyrus
sativus), Lentil (Lens culinaris), Mung bean sprouts (Vigna
radiata), Okra (Abelmoschus esculentus), Pea (Pisum sativum),
Peanut (Arachis hypogaea), Pigeon pea (Cajanus cajan), Rice bean
(Vigna umbellatta), Soybean (Glycine max), Winged bean
(Psophocarpus tetragonolobus), and Yardlong bean (Vigna unguiculata
subsp. sesquipedalis).
[0144] In yet another embodiment, the agricultural product is a
bulb vegetable and/or stem vegetable, as exemplified by, but not
limited to, Asparagus (Asparagus officinalis), Celery (Apium
graveolens), Elephant Garlic (Allium ampeloprasum var.
ampeloprasum), Florence fennel (Foeniculum vulgare var. dulce),
Garlic (Allium sativum), Kohlrabi (Brassica oleracea Gongylodes
group), Leek (Allium porrum), Onion (Allium cepa), Prussian
asparagus (Ornithogalum pyrenaicum), Shallot (Allium cepa
Aggregatum group), Welsh onion (Allium fistulosum), and Wild leek
(Allium tricoccum).
[0145] In a further embodiment, the agricultural product is a root
vegetable and/or tuberous vegetable, as exemplified by, but not
limited to, Bamboo shoot, Beetroot (Beta vulgaris subsp. vulgaris),
Black cumin (Bunium persicum), Broadleaf arrowhead (Sagittaria
latifolia), Carrot (Daucus carota), Cassava (Manihot esculenta),
Chinese artichoke (Stachys affinis), Daikon (Raphanus sativus
Longipinnatus group), Earthnut pea (Lathyrus tuberosus), Ginger
(Zingiber officinale), Hamburg parsley (Petroselinum crispum var.
tuberosum), Jerusalem artichoke (Helianthus tuberosus), Jicama
(Pachyrhizus erosus), Parsnip (Pastinaca sativa), Potato (Solanum
tuberosum), Prairie turnip (Psoralea esculenta), Radish (Raphanus
sativus), Rutabaga (Brassica napus Napobrassica group), Sweet
Potato (Kumara), Taro (Colocasia esculenta), Turnip (Brassica rapa
Rapifera group), Wasabi (Wasabia japonica), Water chestnut
(Eleocharis dulcis), and Yam (Dioscorea spp.).
[0146] In another embodiment, the agricultural product is a sea
vegetable, as exemplified by, but not limited to, Dabberlocks or
badderlocks (Alaria esculenta), Dulse (Palmaria palmata), Hijiki
(Hizikia fusiformis), Kombu (Laminaria japonica), Mozuku
(Cladosiphon okamuranus), Sea grape (Caulerpa spp.), and Sea
lettuce (Ulva lactuca).
[0147] In other embodiments, the agricultural product is a member
of the Rosaceae family, as exemplified by, but not limited to,
Apple and crabapple (Malus), Hawthorn (Crataegus and Rhaphiolepis),
Loquat (Eryobotrya japonica), Pear, European and Asian species
(Pyrus), Quince (Cydonia oblonga and Chaenomeles), Apricot (Prunus
armeniaca or Armeniaca vulgaris), Cherry (Prunus avium), Peach
(Persica, vulgaris), Plum (Prunus salicina), and Strawberry
(Fragaria ananassa).
[0148] In another embodiment, the agricultural product is a bramble
fruit, as exemplified by, but not limited to, Blackberry (genus
Rubus) and Raspberry (genus Rubus), or is a berry, as exemplified
by, but not limited to, Bilberry or whortleberry (Vaccinium spp.),
Blueberry (Vaccinium spp.), Cranberry (Vaccinium spp.), Huckleberry
(Vaccinium spp.), Barberry (Berberis; Berberidaceae), Currant
(Ribes spp.; Grossulariaceae) including red, black, and white
types, Elderberry (Sambucus; Caprifoliaceae), Gooseberry (Ribes
spp.; Grossulariaceae), Hackberry (Celtis spp.; Cannabaceae),
Honeysuckle, (Lonicera spp.; Caprifoliaceae), and Mulberry (Morus
spp.; Moraceae).
[0149] In a further embodiment, the agricultural product is an
Asian fruit, as exemplified by, but not limited to, Coconut (Cocos
spp.; Arecaceae), Kiwifruit or Chinese gooseberry (Actinidia spp.;
Actinidiaceae), Persimmon (aka Sharon Fruit) (Diospyros kaki;
Ebenaceae), Rhubarb (Rheum rhaponticum; Polygonaceae), and
Pineapple (Ananas comosus).
[0150] In another embodiment, the agricultural product is a North
American fruit, as exemplified by, but not limited to, American
grape, (Vitis labrusca; Vitaceae), American persimmon (Diospyros
virginiana; Ebenaceae), Beach Plum (Prunus maritima; Rosaceae),
Blueberry (Vaccinium, sect. Cyanococcus; Ericaceae), Persimmon
(Diospyros virginiana; Ebenaceae), and Saw Palmetto (Serenoa
repens; Ericaceae).
[0151] In yet another embodiment, the agricultural product is a
citrus fruit, as exemplified by, but not limited to, Blood Orange,
Citron (Citrus medica), Clementine (Citrus reticulata var.
Clementine), Grapefruit (Citrus paradisi), Kumquat (Fortunella),
Lemon (Citrus limon), Key Lime (Citrus aurantifolia), Mandarin
(Citrus reticulata), Orange, (Citrus sinensis), Pomelo (also known
as the shaddock) (Citrus maxima), and Sweet Lemon (Citrus
limetta).
[0152] In further embodiments, the agricultural product is meat,
such as raw and/or cooked beef, pork, and lamb, exemplified by beef
steak, beef hamburger, pork chop, pork sausage, lamb shank, and
bacon.
[0153] In other embodiments, the agricultural product is poultry,
such as raw and/or cooked turkey, chicken, and duck, including
whole poultry and poultry parts, e.g., breast, thigh, drumstick,
and poultry products e.g., poultry eggs, and egg shells.
[0154] In yet other embodiments, the agricultural product is
seafood such as raw (e.g., sushi) and/or cooked fish (e.g., salmon,
bass, tuna), including whole fish and fish parts, e.g., steak,
fillet, as well as shellfish (e.g., mussels, clams, oysters, and
shrimp).
[0155] In further embodiments, the agricultural product is a fresh
or cooked seed or nut, as exemplified by, but not limited to,
almonds (Prunus communis), walnuts (Juglans nigra), sunflower
(Helianthus, annuus), and pumpkin (Cucurbita pepo).
[0156] Other embodiments of agricultural products include fresh
and/or dried herb such as parsley (Carum petroselinum), cumin
(Cuminum cyminum), sweet basil (Ocimum basilicum), rosemary
(Rosmarinus officinalis), and mint (Mentha spp).
[0157] Further embodiments of agricultural product include animal
hide (e.g., cow hide and pig hide), and poultry feathers (e.g.,
chicken feathers, turkey feathers and goose feathers).
[0158] Other surfaces that may be treated with the invention's
compositions include skin (e.g., human skin, cow udders, cow teats,
etc.) and inanimate objects (e.g., door handles, door knobs, door
frames, table surfaces, stair railings).
[0159] Experimental
[0160] The following examples serve to illustrate certain preferred
embodiments and aspects of the present invention and are not to be
construed as limiting the scope thereof.
EXAMPLE 1
[0161] Synergistic Antimicrobial Activity
[0162] This example provides the results for antimicrobial activity
of components of exemplary invention's compositions when the
components were used alone (Tables 3A and 3B), or as a combination
of two components (Tables 3A and 4), or of at least three
components (Tables 3A and 5).
[0163] Produce was rinsed with sterile water before refrigerating.
Produce was dip-inoculated and let stand for 24 hrs prior to
component testing. 25-gram samples were pureed with 475 grams of
sterile water (20 fold dilution). Pureed samples were spread-plated
on pre-made Mac Conkey with mug agar plates #221172 from Becton,
Dickinson Biosciences and incubated at about 44.degree. F. for
24-30 hours. Colonies were counted using colony counter Bantex 920A
with UV light 366 nM. The results are shown in Table 3A.
TABLE-US-00003 TABLE 3A Antimicrobial activity (log reduction) of
components used singly, in combination of two components, in
combination of three components, and in combination of four
components. E. Coli (ATCC 29522) inoculum 5.45 log Strawberry 25
grams Spinach leaf 25 grams Component.sup.(a) 2 min..sup.(b) 30
min..sup.(b) 180 min..sup.(b) 2 min..sup.(b) 30 min..sup.(b) 180
min..sup.(b) Single Components Propylene Glycol 1.83 2.24 2.55 1.71
2.1 2.4 (0.03 wgt. %) Phosphoric Acid 2.15 2.17 2.62 2.00 2.1 2.55
(0.3 wgt. %) Hydrogen 2.0 2.66 2.99 1.8 2.44 2.77 Peroxide (0.2
wgt. %) Sodium Lauryl 1.2 1.5 1.89 1.0 1.6 1.78 Sulfate (0.0005
wgt. %) Potable Water 0.91 0.98 1.05 0.78 0.82 0.88 alone
(33.degree. F., 100 wgt. %) Two Components Propylene Glycol 2.99
4.01 5.45 2.78 3.89 5.45 (0.03 wgt. %) and Phosphoric Acid (0.3
wgt. %) Three Components Propylene Glycol 3.34 4.45 5.45 3.12 4.12
5.45 (0.03 wgt. %) and Phosphoric Acid (0.3 wgt. %) and Hydrogen
Peroxide (0.2 wgt. %) Four Components Propylene Glycol 4.12 5.11
5.45 3.98 4.99 5.45 (0.03 wgt. %) and Phosphoric Acid (0.3 wgt. %)
and Hydrogen Peroxide (0.2 wgt. %) and Sodium Lauryl Sulfate
(0.0005 wgt. %).sup.(c) .sup.(a)Component was dissolved in potable
water to make a total of 100 g. The component and sterile water
were chilled to 34.degree. F. .sup.(b)Fruit and vegetable were
chilled to 40.degree. F. and washed in the solution containing the
listed component for the period indicated. Testing was done at
ambient temperature of about 68.degree. F. .sup.(c)The solution
containing Propylene Glycol 0.03 wgt. %, ortho-Phosphoric Acid 0.3
wgt. %, Hydrogen Peroxide 0.2 wgt. % and Sodium Lauryl Sulfate
0.0005 wgt. % is an exemplary composition of solution D of Table 1.
It was made by using a concentrated solution of Propylene Glycol 3
wgt. %, ortho-Phosphoric Acid 30 wgt. %, Hydrogen Peroxide 20 wgt.
% and Sodium Lauryl Sulfate 0.05 wgt. % diluted 100 to 1.
TABLE-US-00004 TABLE 3B Antimicrobial activity (log reduction) of
single components E. Coli (ATCC 43888) inoculum 6.87-log Strawberry
25 grams Spinach leaf 25 grams Component.sup.(a) 2 min..sup.(b) 30
min..sup.(b) 180 min..sup.(b) 2 min..sup.(b) 30 min..sup.(b) 180
min..sup.(b) Propylene Glycol (2 wgt. 2.23 2.69 4.47 2.44 2.54 3.98
%) Acetic Acid (1 wgt. 2.52 2.73 4.45 2.5 2.61 4.01 %) Citric Acid
(1 wgt. 2.25 2.68 4.43 2.11 2.34 4.09 %) Phosphoric Acid (1 wgt.
2.58 2.87 4.22 2.5 2.71 4.13 %) Hypochlorous Acid 2.45 3.45 5.69
2.32 3.4 5.01 (0.01 wgt. %) Bromine (0.08 wgt. 2.32 3.32 4.89 2.11
3.28 4.44 %) Hydrogen Peroxide 2.11 2.69 4.65 2.01 2.6 3.89 (2 wgt.
%) Ozone (0.001 wgt. 2.89 3.12 4.12 2.34 2.58 3.63 %) Sodium Lauryl
1.89 2.69 3.56 1.77 2.29 3.26 Sulfate (0.5 wgt. %) (CAS
009004-82-4) Octyl Phenol 1.05 2.78 3.9 1.15 2.47 3.11 Ethoxylate
(0.5 wgt. %) Potable Water alone 0.98 1.04 1.2 0.68 1 1.1
.sup.(a)Component was dissolved in potable water to make a total of
100 g and tested at about 33.degree. F. .sup.(b)Fruit and vegetable
were washed in the solution containing the listed component for the
period indicated.
TABLE-US-00005 TABLE 4 Antimicrobial activity (log reduction) of a
combination of two E. Coli (ATCC 43888) inoculum 6.87-log
Strawberry 25 grams Spinach leaf 25 grams Component.sup.(a) 2
min..sup.(b) 30 min..sup.(b) 180 min..sup.(b) 2 min..sup.(b) 30
min..sup.(b) 180 min..sup.(b) Hypochlorous 2.68 3.74 4.98 2.48 3.54
4.78 Acid 0.01 wgt. % & Acetic Acid 1 wgt. % Hypochlorous 2.55
3.51 4.66 2.25 3.3 4.48 Acid 0.01 wgt. % & Citric Acid 1 wgt. %
Hypochlorous 2.81 3.94 5.13 2.61 3.74 4.9 Acid 0.01 wgt. % &
Phosphoric Acid 1 wgt. % Bromine 0.08 wgt. 2.71 3.39 4.91 2.51 3.1
4.7 % & Acetic Acid 1 wgt. % Bromine 0.08 wgt. 2.44 3.64 4.71
2.52 3.54 4.71 % & Citric Acid 1 wgt. % Bromine 0.08 wgt. 2.82
3.93 4.74 2.42 3.63 4.88 % & Phosphoric Acid 1 wgt. % Hydrogen
2.51 3.56 4.89 2.43 3.5 4.77 Peroxide 2 wgt. % & Acetic Acid 1
wgt. % Hydrogen 2.54 3.57 4.85 2.4 3.68 4.85 Peroxide 2 wgt. %
& Citric Acid 1 wgt. % Hydrogen 2.72 3.81 4.99 2.69 3.71 4.99
Peroxide 2 wgt. % & Phosphoric Acid 1 wgt. % Ozone 0.001 wgt.
2.49 3.79 4.96 2.65 3.5 4.9 % & Acetic Acid 1 wgt. % Ozone
0.001 wgt. 2.48 3.77 4.96 2.48 3.57 4.7 % & Citric Acid 1 wgt.
% Ozone 0.001 wgt. 2.47 3.78 4.97 2.5 3.7 4.78 % & Phosphoric
Acid 1 wgt. % Hypochlorous 2.41 3.31 4 2.61 3.41 4.1 Acid 0.01 wgt.
% & Sodium Lauryl Sulfate 0.5 wgt. % Hypochlorous 2.56 3.41
4.11 2.4 3.21 4 Acid 0.01 wgt. % & Octyl Phenol Ethoxylate 0.5
wgt. % Bromine 0.08 wgt. 2.31 2.98 3.98 2.3 2.88 3.99 % &
Sodium Lauryl Sulfate 0.5 wgt. % Bromine 0.08 wgt. 2.45 2.99 4 2.4
2.9 3.78 % & Octyl Phenol Ethoxylate 0.5 wgt. % Hydrogen 2.34
2.89 3.57 2.35 2.88 3.55 Peroxide 2 wgt. % & Sodium Lauryl
Sulfate 0.5 wgt. % Hydrogen 2.46 2.93 3.91 2.33 2.78 3.81 Peroxide
2 wgt. % & Octyl Phenol Ethoxylate 0.5 wgt. % Ozone 0.001 wgt.
2.56 3.51 3.98 2.56 3.31 3.78 % & Sodium Lauryl Sulfate 0.5
wgt. % Ozone 0.001 wgt. 2.54 3.41 3.9 2.45 3.22 3.39 % & Octyl
Phenol Ethoxylate 0.5 wgt. % .sup.(a)Component was dissolved in
potable water to make a total of 100 g and tested at about
33.degree. F. .sup.(b)Fruit and vegetable were washed in the
solution containing the listed component for the period
indicated.
TABLE-US-00006 TABLE 5 Antimicrobial activity (log reduction) of a
combination of at least three components. E. Coli (ATCC 43888)
inoculum 6.57-log Strawberry 25 grams Spinach leaf 25 grams 2 30
180 2 30 180 Component .sup.(a) min..sup.(b) min..sup.(b)
min..sup.(b) min..sup.(b) min..sup.(b) min..sup.(b) (a1) 4 5.1 6.57
4.5 5.2 6.57 Hypochlorous Acid 0.01 wgt. % & Phosphoric Acid 1
wgt. % & Propylene Glycol 2 wgt. % (a2) 3.9 4.48 6.49 4 4.58
6.49 Bromine 0.08 wgt. % & Phosphoric Acid 1 wgt. % &
Propylene Glycol 2 wgt. % (a3) 3.98 4.22 6.5 3.99 4.32 6.5 Hydrogen
Peroxide 2 wgt. % & Phosphoric Acid 1 wgt. % & Propylene
Glycol 2 wgt. % (a4) 3.99 4.77 6.49 3.99 4.76 6.48 Ozone 0.001 wgt.
% & Phosphoric Acid 1 wgt. % & Propylene Glycol 2 wgt. %
(a5) 4.12 5.44 6.57 4.22 5.3 6.57 Hypochlorous Acid 0.01 wgt. %
& Bromine 0.05 wgt. % & Phosphoric Acid 1 wgt. % &
Propylene Glycol 2 wgt. % .sup.(a) Components were dissolved in
potable water to make a total of 100 g and tested at about
33.degree. F. .sup.(b)Fruit and vegetable were washed in the
solution containing the listed components for the period
indicated.
[0164] In the above Tables 3-5, the antimicrobial activity is
reported as a log reduction which was determined by calculating the
difference between the log10 of the initial inoculum count and the
log10 of the inoculum's count after exposure to the listed
components for about 2 minutes, about 30 minutes, or about 180
minutes, at about 33.degree. F.
[0165] To determine bacterial kill-rate or log10 reduction, a 0.1
ml aliquot of a bacterial culture suspension incubated for about 24
hours in tryptic soy broth having an initial inoculum count of
between about 106 to about 108 cells/ml was added to a test sample
of either fruits (strawberry 25 grams) or vegetables (spinach 25
grams) at about 33.degree. F. In this example, culture suspensions
were prepared from E. coli (ATCC 43888). After about two minutes,
thirty minutes or one hundred eighty minutes of this treatment,
using the USDA recommended procedure, the inoculated material was
placed into a 225 ml solution of EC Medium, with mug, in a
stomacher bag and thoroughly mixed for a minimum of two minutes and
incubated at 35.degree. C. for twenty four hours. Next, the
cultures were diluted 10-fold in Butterfield's Phosphate Diluent
and 0.1 ml of the dilutions were inoculated using the spread plate
technique onto MacConkey Sorbitol Agar (MSA). Then the plates were
incubated for twenty four hours at 42.degree. C. Plates were then
removed and examined for the number of colonies.
[0166] The data demonstrate synergy of the invention's components.
For example, Table 3B shows that 180 minute treatment of spinach
with each of hypochlorous acid 0.01 wgt. %, phosphoric acid 1 wgt.
% and propylene glycol 2 wgt. % alone in water caused a 5.01-log,
4.13-log, and 3.98-log reduction, respectively, in a 6.87-log E.
coli inoculum. Table 4 shows similarly low antimicrobial activity
by a combination of two components of hypochlorous acid 0.01 wgt. %
and phosphoric acid 1 wgt. % which resulted in a 4.9-log reduction
in a 6.87-log E. coli inoculum. In contrast, Table 5 shows that 180
minute treatment of spinach with the combination "al" that
contained hypochlorous acid 0.01 wgt. %, phosphoric acid 1 wgt. %,
and propylene glycol 2 wgt. % resulted in a 100% reduction in a
6.57-log E. coli inoculum.
[0167] Similarly, Table 3B shows that 180 minute treatment of
spinach with each of bromine 0.08 wgt. %, phosphoric acid 1 wgt. %
and propylene glycol 2 wgt. % alone caused a 4.44-log, 4.13-log,
and 3.98-log reduction, respectively, in a 6.87-log E. coli
inoculum. Table 4 shows similarly low antimicrobial activity by a
combination of two components of bromine 0.08 wgt. % and phosphoric
acid 1 wgt. % which resulted in a 4.88-log reduction in a 6.87-log
E. coli inoculum. In contrast, Table 5 shows that 180 minute
treatment of spinach with the combination "a2" that contained
bromine 0.08 wgt. %, phosphoric acid 1 wgt. % and propylene glycol
2 wgt. % resulted in a 6.49-log reduction in a 6.57-log E. coli
inoculum.
[0168] In yet another example, Table 3B shows that 180 minute
treatment of spinach with each of hypochlorous acid 0.01 wgt. %,
bromine 0.05 wgt. %, phosphoric acid 1 wgt. %, and propylene glycol
2 wgt. % alone caused a 5.01-log, 4.44-log, 4.13-log and 3.98-log
reduction, respectively, in a 6.87-log E. coli inoculum. Table 4
shows similarly low antimicrobial activity by a combination of two
components of bromine 0.08 wgt. % and phosphoric acid 1 wgt. %
which resulted in a 4.88-log reduction in a 6.87-log E. coli
inoculum. In contrast, Table 5 shows that 180 minute treatment of
spinach with the combination "a5" that contained hypochlorous acid
0.01 wgt. %, bromine 0.05 wgt. %, phosphoric acid 1 wgt. %, and
propylene glycol 2 wgt. % resulted in a 100% reduction in a
6.57-log E. coli inoculum.
EXAMPLE 2
[0169] Antimicrobial Activity Using Agricultural Products and
Inanimate Objects
[0170] This example shows the results of treating a variety of
agricultural products, skin, and inanimate objects with the
invention's exemplary compositions "x1" to "x15" that are described
above in Table 2.
TABLE-US-00007 TABLE 6 Antimicrobial Activity Of Exemplary
Compositions Of The Invention Reference Treatment Log Number of
Method.sup.(a) Log Reduction Exemplary (see Table 7 Microbe Microbe
in Material Composition for details) Tested.sup.(b) Inoculum
Microbe.sup.(c) Notes Botanical Leafy vegetables Bok choy x1 t1 b2
6.88 >6.7 (Brassica rapa (E. coli Chinensis generic) group) x3
t3 b4 8.78 <2.3 Composition (Lactobacillus x3 diluted sp.) 100
to 1 Cabbage x1 t1 b2 6.88 >6.8 (Brassica (E. coli oleracea
generic) Capitata group) x3 t3 b4 8.78 <1 Composition
(Lactobacillus x3 diluted sp.) 100 to 1 x15 t2 b1 6.87 >6.7
Composition (E. coli) x15 diluted 10 to 1 Dandelion x1 t1 b2 6.88
>6.8 (Taraxacum (E. coli officinale) generic) x3 t3 b4 8.78
<2 Composition (Lactobacillus x3 diluted sp.) 100 to 1 Lettuce
x1 t1 b2 6.88 >6.8 (Lactuca sativa) (E. coli generic) x3 t3 b4
8.78 <3 Composition (Lactobacillus x3 diluted sp.) 100 to 1 x4
t2 b3 4.21 >4 (Staphylococcus aureus) x4 t2 b5 5.45 >5
(Salmonella enterica) Mustard x1 t1 b2 6.88 >6.8 (Sinapis alba)
(E. coli generic) x3 t3 b4 8.78 <1 Composition (Lactobacillus x3
diluted sp.) 100 to 1 Napa/Chinese x1 t1 b2 6.88 >6.6 Cabbage
(E. coli (Brassica rapa generic) Pekinensis group) x3 t3 b4 8.78
<1 Composition (Lactobacillus x3 diluted sp.) 100 to 1 Spinach
x1, t1 b2 6.88 >6.7 (Spinacia (E. coli oleracea) generic) x3 t3
b4 8.78 <2 Composition (Lactobacillus x3 diluted sp.) 100 to 1
x11 t3 b1 6.87 >6.5 Composition (E. coli) x11 diluted 200 to 1
x7 t2 b5 5.45 >5 (Salmonella enterica) Swiss chard x1 t1 b2 6.88
>6.8 (Beta vulgaris (E. coli subsp. cicla var. generic)
flavescens) x3 t3 b4 8.78 <1 Composition (Lactobacillus x3
diluted sp.) 100 to 1 x9 t1 b2 6.88 >6.6 (E. coli generic) x1 t1
p1 3.1 <1 (bacteriophage T4) Botanical Fruiting and flowering
vegetables Avocado x1 t3 b1 6.87 >6.7 (Persea (E. coli)
americana) x2 t1 b1 6.87 >6 (E. coli) Bell pepper x4 t3 b1 6.87
>6.8 Composition (Capsicum (E. coli) x4 diluted annuum) 100 to 1
x9 t1 b1 6.87 >6.7 (E. coli) Globe x5 t3 b1 6.87 >6.6
Artichoke (E. coli) (Cynara scolymus) x7 t2 b1 6.87 >6.8 (E.
coli) Tomato x1 t1 b2 6.88 >6.8 (Solanum (E. coli lycopersicum)
generic) x2 t2 b5 5.45 <2 (Salmonella enterica) x11 t3 m1 5.5
>4 Composition (Stachybotrys x11 diluted chartarum) 100 to 1 x1
t2 b2 6.88 >6.6 (E. coli generic) x13 t3 ml 5.5 >5
Composition (Stachybotrys x13 diluted chartarum) 100 to 1 Botanical
Podded vegetables Mung bean x1 t1 b1 6.87 >6.5 sprouts (Vigna
(E. coli) radiata) x4 t3 b1 6.87 >6.1 Composition (E. coli) x4
diluted 100 to 1 Botanical Bulb and stem vegetables Asparagus x10
t3 b1 6.87 >6.5 (Asparagus (E. coli) officinalis) Botanical Root
and tuberous vegetables Carrot (Daucus x10 t3 b1 6.87 >6.6
carota) (E. coli) Botanical Rosaceae family Apple and x10 t3 b1
6.87 >6.6 crabapple (E. coli) (Malus) Peach x10 t3 b1 6.87
>6.5 (Persica, (E. coli) vulgaris) Strawberry x1 t3 b1 6.87
>6.7 (Fragaria (E. coli) ananassa) x4 t1 b2 6.88 >6.6
Composition (E. coli x4 diluted generic) 100 to 1 x11 t2 m1 5.5
>4 Composition (Stachybotrys x11 diluted chartarum) 50 to 1
Botanical Bramble fruits Blackberry, x11 t3 b1 6.87 >6.8
Composition (genus Rubus) (E. coli) x11 diluted 100 to 1 Botanical
Berries Cranberry x11 t3 b1 6.87 >6.7 Composition (Vaccinium (E.
coli) x11 diluted spp.) 100 to 1 Botanical Fruit Asian Pineapple
cut x11 t3 b1 6.87 >5 Composition into (E. coli) x11 diluted
slices(Ananas 500 to 1 comosus) Botanical Fruit North American
Blueberry x11 t3 b1 6.87 >6 Composition (Vaccinium, (E. coli)
x11 diluted sect. 100 to 1 Cyanococcus; Ericaceae) Meat Raw Beef x1
t8 b1 6.87 >5 Hamburger (E. coli) Pork Chop x11 t9 b1 6.87 >6
Composition (E. coli) x11 diluted 100 to 1 Bacon x1 t8 b1 6.87
>6.5 (E. coli) Poultry Raw Turkey breast x11 t6 b1 6.87 >6
Composition (E. coli) x11 diluted 100 to 1 Chicken breast x1 t6 b1
6.87 >6.8 (E. coli) Fish Raw Salmon steak x1 t13 b1 6.87 >6.8
(E. coli) Bass steak x12 t13 b1 6.87 >6.8 (E. coli) tuna
processed x12 t13 b1 6.87 >6 raw (E. coli) Shellfish Clams raw
x1 t14 b1 6.87 >6 (E. coli) shrimp raw not x1 t14 b1 6.87
>6.5 frozen (E. coli) Seeds and Nuts fresh (not dried) Almonds
x12 t11 m1 5.5 >4 (Prunus (Stachybotrys communis) chartarum) x12
t11 f1 2.3 >1.8 (Aspergillus flavus) Eggs (topical Shells)
Chicken fresh x15 t4 b1 6.87 >6 eggs (E. coli) Herbs fresh
parsley (Carum x1 t1 b1 6.87 >6 petroselinum) (E. coli) Herbs
Dried parsley (Carum x13 t15 b1 6.87 >6 Composition
petroselinum) (E. coli) x13 diluted 200 to 1 Animal Hides Cow Hide
x11 t10 b1 6.87 >6 Composition (E. coli) x11 diluted 50 to 1
Animal feathers Chicken x11 t7 b5 5.45 >5 feathers (Salmonella
enterica) Human skin Male age 46, x10 t19 skin test -- forearm Male
age 78, x10 t19 b3 4.21 -- top left hand (Staphylococcus aureus)
Structures Pine wood x5 t18 m1 5.5 >3.5 sanded (Stachybotrys 6''
.times. 6'' chartarum) x9 t18 b1 6.87 >5 (E. coli) x14 t18 b2
6.88 >5
(E. coli generic) Teflon sheet x5 t18 m1 5.5 >5 6'' .times. 6''
(Stachybotrys chartarum) x9 t18 b1 6.87 >6 (E. coli) x14 t18 b2
6.88 >6 (E. coli generic) Ultra high x5 t18 m1 5.5 >5
molecular (Stachybotrys weight chartarum) (UHMW) polyethylene sheet
6'' .times. 6'' x9 t18 b1 6.87 >6 (E. coli) x14 t18 b2 6.88
>6 (E. coli generic) ceramic tile x13 t17 m1 5.5 5.5 Composition
3'' .times. 3'' (Stachybotrys x13 diluted chartarum) 10 to 1 x13
t11 v1 2.4 2.4 Composition (Rhinovirus) x13 diluted 10 to 1
stainless steel x13 t11 m1 5.5 5.5 Composition 6'' .times. 6''
(Stachybotrys x13 diluted chartarum) 10 to 1 x13 t17 v1 2.4 2.4
Composition (Rhinovirus) x13 diluted 10 to 1 cement block x13 t11
m1 5.5 5.5 Composition 6'' .times. 6'' (Stachybotrys x13 diluted
chartarum) 10 to 1 x13 t11 v1 2.4 2.4 Composition (Rhinovirus) x13
diluted 10 to 1 Glass sheet x13 t11 m1 5.5 5.5 Composition 6''
.times. 6'' (Stachybotrys x13 diluted chartarum) 10 to 1 x13 t11 v1
2.4 2.4 Composition (Rhinovirus) x13 diluted 10 to 1 Painted
(acrylic x13 t18 m1 5.5 >4 Composition paint) wood (Stachybotrys
x13 diluted 6'' .times. 6'' chartarum) 10 to 1 x13 t18 v1 2.4 >2
Composition (Rhinovirus) x13 diluted 10 to 1 ICE with x1 t16 b1
6.87 >6 shrimp (E. coli) x2 t16 b1 6.87 >6.5 (E. coli) Plant
specific Tomato leaf x11 t1 f2 3.2 >2 Composition (Solanum
(Septoria x11 diluted lycopersicum) lycopersici) 50 to 1 x11 t1 n1
2.3 >1.8 Composition (Aphelenchoides x11 diluted fragariae) 50
to 1 Poultry x10 t7 b5 5.45 >5 feathers (Salmonella enterica)
Reverse x3 t21 b2 6.88 6.88 Composition Osmosis (RO) (E. coli x3
diluted membrane generic) 200 to 1 .sup.(a)All tests were performed
according to FDA specifications. .sup.(b)Microbes: E. coli (ATCC
43888), E. coli generic (ATCC 25922), Staphylococcus aureus (ATCC
25923), Salmonella enterica (ATCC 10708), Lactobacillus sp. (ATCC
55326). Aspergillus flavus (ATCC 15517) is an almond fungus;
Septoria lycopersici (ATCC Q99324) is a tomato leaf fungus;
Stachybotrys chartarum (ATCC 9182) is black mold (toxic);
bacteriophage T4 (ATCC 35060-B4); Aphelenchoides fragariae (ATCC
12974) is a nematode that destroys plant crops, necessitating
burning of crops to eradicate the nematode; rhinovirus (ATCC
vr1110) causes head cold in humans. .sup.(c)< means less than;
> means more than.
TABLE-US-00008 TABLE 7 Exemplary Treatment Methods Used To Obtain
Some Of The Data In Table 6 Treatment Reference Treatment
Method.sup.(a) t1 Fruits and vegetables may be dipped, sprayed,
fogged or aerosoled at temperatures equal to or above 40 degrees F.
as a pretreatment before the final processing wash. Contact time
with the invention's composition may be 45 minutes or longer. t2
Fruits and vegetables may be dipped, sprayed, fogged or aerosoled
at temperatures equal to or below 40 degrees F. as a pretreatment
before the final processing wash. Contact time with the invention's
composition may be 100 minutes or longer. t3 Fruits and vegetables
may be liquid conveyed in composition at temperatures equal to or
below 40 degrees F. as a final processing wash. Contact time with
the invention's composition may be 100 minutes or longer. t4 Raw
eggs may be dipped, sprayed, fogged or aerosoled at temperatures
equal to or above 34 degrees F. as a final treatment before
packaging. Contact time with the invention's composition may be 45
minutes or longer. t5 Poultry with feathers may be dipped, sprayed,
fogged or aerosoled at temperatures equal to or above 70 degrees F.
as a pretreatment before de- feathering. Contact time with the
invention's composition may be 15 minutes or longer. Feathers will
have reduced numbers of microbes and can be dried and ready for
processing. for use as fertilizer. t5 Poultry with feathers may be
dipped, sprayed, fogged or aerosoled at temperatures equal to or
above 70 degrees F. as a pretreatment before de- feathering.
Contact time with the invention's composition may be 15 minutes or
longer. Feathers will have reduced numbers of microbes and can be
dried and ready for processing. t6 Poultry that has been
de-feathered may be dipped, sprayed, fogged or aerosoled at
temperatures equal to or above 34 degrees F. as a final treatment
before packaging. Contact time with the invention's composition may
be 45 minutes or longer. t6 Poultry that has been de-feathered may
be dipped, sprayed, fogged or aerosoled at temperatures equal to or
above 34 degrees F. Contact time with the invention's composition
may be 45 minutes or longer. t7 Detached Poultry feathers may be
dipped, sprayed, fogged or aerosoled at temperatures equal to or
above 70 degrees F. as a final treatment before palletizing further
use. Contact time with the invention's composition may be 5 minutes
or longer. Feathers will have reduced numbers of microbes and
residual activity even after can be dried and are ready drying. for
processing for use as fertilizer. t7 Detached poultry feathers may
be dipped, sprayed, fogged or aerosoled at temperatures equal to or
above 70 degrees F. as a final treatment before further use.
Contact time with the invention's composition may be 5 minutes or
longer. Feathers will have reduced numbers of microbes and residual
activity even after drying. t8 Meats other than poultry, may be
dipped, sprayed, fogged or aerosoled at temperatures equal to or
above 34 degrees F. as a final wash. Contact time with the
invention's composition may be 45 minutes or longer. t9 Meats other
than poultry, maybe dipped, sprayed, fogged or aerosoled at
temperatures equal to or above 34 degrees F. as a final wash.
Contact time with the invention's composition may be 45 minutes or
longer. The composition is preferably washedrinsed off. t9 Meats
other than poultry, maybe dipped, sprayed, fogged or aerosoled at
temperatures equal to or above 34 degrees F. as a final wash.
Contact time with the invention's composition may be 45 minutes or
longer. The composition is preferably rinsed off. t10 Hides (animal
skins) may be dipped, sprayed, fogged or aerosoled at temperatures
equal to or above 34 degrees F. as a pretreatment. Contact time
with the invention's composition may be 5 minutes or longer.
Animals may be alive or dead for this process. The composition may
be left on to dry before the remainder of the processing steps. t11
Seeds and nuts may be dipped, sprayed, fogged or aerosoled at
temperatures equal to or above 34 degrees F. as a final wash.
Contact time with the invention's composition may be 45 minutes or
longer. t12 Seeds and nuts may be dipped, sprayed, fogged or
aerosoled at temperatures equal to or above 34 degrees F. as a
final wash. Contact time with the invention's composition may be 15
minutes or longer. The composition is preferably washedrinsed off.
t12 Seeds and nuts may be dipped, sprayed, fogged or aerosoled at
temperatures equal to or above 34 degrees F. as a final wash.
Contact time with the invention's composition may be 15 minutes or
longer. The composition is preferably rinsed off. t13 Fish may be
dipped, sprayed, fogged or aerosoled at temperatures equal to or
above 34 degrees F. as a final wash. Contact time with the
invention's composition may be 15 minutes or longer. t14 Shellfish
may be dipped, sprayed, fogged or aerosoled at temperatures equal
to or above 34 degrees F. as a final wash. Contact time with the
invention's composition may be 15 minutes or longer. t15 Dried
goods may be sprayed, fogged or aerosoled at temperatures equal to
or above 34 degrees F. as a final wash. Contact time with the
invention's composition may be Contact time with the invention's
composition may be 115 minutes or longer. t15 Dried goods may be
sprayed, fogged or aerosoled at temperatures equal to or above 34
degrees F. as a final wash. Contact time with the invention's
composition may be 15 minutes or longer. t16 Ice may be formed with
the composition that is to be used as an aid for cold storage
transport and microbial reduction. The shrimp were dipped into a
solution containing a bacterial inoculum, and then surrounded for
30 minutes at ambient temperature (about 62.degree. F.) by the
invention's compositions that had previously been frozen into ice
cubes (about 29.degree. F.). Materials surrounded by the ice could
be rinsed off, or left without washing off to allow formation of a
bio-thin, adherent film that provides continued antimicrobial
activity. t16 Ice may be formed with the composition that is to be
used as an aid for cold storage transport and microbial reduction.
The shrimp were dipped into a solution containing a bacterial
inoculum, and then surrounded for 30 minutes at ambient temperature
(about 62.degree. F.) by the invention's compositions that had
previously been frozen into ice cubes (about 29.degree. F.).
Materials surrounded by the ice could be rinsed off, or left
without rinsing to allow formation of a thin, adherent film that
provides continued antimicrobial activity. t17 Structures may be
dipped, sprayed, fogged, aerosoled, autoclaved or scrubbed in at
temperatures equal to or greater than 200 degrees F. as a final
wash. Contact time with the invention's composition may be 2
minutes or longer. The composition is preferably left on the
surface without washing off to allow formation of a bio-thin,
adherent film that provides continued antimicrobial activity t17
Structures may be dipped, sprayed, fogged, aerosoled, autoclaved or
scrubbed at temperatures equal to or greater than 200 degrees F. as
a final wash. Contact time with the invention's composition may be
2 minutes or longer. The composition is preferably left on the
surface without rinsing to allow formation of a thin, adherent film
that provides continued antimicrobial activity t18 Structures may
be dipped, sprayed, fogged, aerosoled or scrubbed in at
temperatures equal to or greater than 34 degrees F. as a final
wash. Contact time with the invention's composition may be 10
minutes or longer. t18 Structures may be dipped, sprayed, fogged,
aerosoled or scrubbed at temperatures equal to or greater than 34
degrees F. as a final wash. Contact time with the invention's
composition may be 10 minutes or longer. t19 Human skin may be
washed at temperatures equal to or above 34 degrees F. as an
antimicrobial wash. Contact time with the invention's composition
may be 2 minutes or longer. Contact time of 4-12 hours did not
result in skin irritation. The moist composition may be lightly
toweled off for a dry appearance. t19 Human skin may be washed at
temperatures equal to or above 34 degrees F. as an antimicrobial
wash. Contact time with the invention's composition may be 2
minutes or longer. Contact time of 4-12 hours did not result in
skin irritation. The moist composition may be lightly toweled for a
dry appearance. t20 Human skin may be washed at temperatures equal
to or above 34 degrees F. as an antimicrobial wash. Contact time
with the invention's composition may be 15 minutes or longer. The
composition is preferably washedrinsed off t20 Human skin may be
washed at temperatures equal to or above 34 degrees F. as an
antimicrobial wash. Contact time with the invention's composition
may be 15 minutes or longer. The composition is preferably rinsed
off t21 The invention's composition was circulated at temperatures
equal to or above 34 degrees F. through the RO membrane for 1
minute using a high pressure multiple centrifuge pump. t21 The
invention's composition was circulated at temperatures equal to or
above 34 degrees F. through a RO membrane for 1 minute using a high
pressure multiple centrifuge pump. .sup.(a)Contact time with the
invention's compositions refers to the period of time the
composition is in contact with a substance before rinsing, drying,
or toweling.
[0171] The above Table 6 shows the antimicrobial activity of the
invention's compositions on bacterial, bacteriophage, viral, fungal
and nematode microbes, using a wide variety of agricultural
products (including leafy vegetables, fruiting and flowering
vegetables, podded vegetables, bulb and stem vegetables, root and
tuberous vegetables, Rosaceae family fruits, Bramble fruits,
berries, Asian fruit, North America fruit, raw meat, raw poultry,
raw seafood, fresh seeds sprouts, nuts, eggs, fresh herbs, dried
herbs, spices, animal hides, feathers) and inanimate objects (such
as those encountered in hospitals, food processing plants,
residential buildings, office buildings, etc.).
[0172] Furthermore, the data demonstrate that, surprisingly, the
invention's compositions are effective antimicrobials even at their
freezing temperatures, as shown by the reduction of E. coli on
shrimp that has been in contact with ice cubes containing the
invention's compositions.
[0173] Moreover, Table 6 demonstrates the surprising differential
effect of the invention's compositions on pathogenic bacteria on
the one hand, and on non-pathogenic bacteria and bacteriophage on
the other hand. For example, the data show higher antimicrobial
activity against both the pathogenic bacteria E. coli
(Gram-negative) and Staphylococcus aureus (Gram-positive) compared
to the non-pathogenic Lactobacillus (Gram-positive) and
bacteriophage T4.
[0174] Table 6 also demonstrates that the invention's compositions
are suitable antimicrobials for application to human skin.
EXAMPLE 3
[0175] Larger Scale Testing of Antimicrobial Activity
[0176] This example was used to demonstrate large batch processing
before cutting or chopping of produce. Testing procedures were done
in accordance to Example 1. Two hundred pounds of each of spinach,
spring mix and iceberg lettuce were inoculated with E. coli. ATCC
Number 43888 and the vegetables were tested using immersion and a
bubbler tank The results are shown in Table 8.
TABLE-US-00009 TABLE 8 Antimicrobial activity (log reduction) of a
combination of three components E. Coli (ATCC 43888) inoculum
5.51-log Spinach leaf Spring Mix.sup.(b) Iceberg Lettuce (200
pounds) (200 pounds) (200 pounds) 2 30 180 2 30 180 2 30 180
Component.sup.(a) min..sup.(c) min..sup.(c) min..sup.(c)
min..sup.(c) min..sup.(c) min..sup.(c) min..sup.(c) min..sup.(c)
min..sup.(c) (a1) 4.1 5 5.51 4.4 5.1 5.51 4.3 5.22 5.51
Hypochlorous Acid 0.01 wgt. % & Phosphoric Acid 1 wgt. % &
Propylene Glycol 2 wgt. % (a2) 3.5 4.58 5.51 3.6 5 5.51 3.44 5 5.51
Hydrogen Peroxide 2 wgt. % & Phosphoric Acid 1 wgt. % &
Propylene Glycol 2 wgt. % .sup.(a)Components were dissolved in 350
liters of potable water and tested at about 33.degree. F.
.sup.(b)Spring mix may contain any combination of two or more of
baby lettuce, greens, endive and radicchio. .sup.(c)The vegetables
were washed in the solution containing the listed components for
the period indicated.
[0177] The results demonstrate that the invention's compositions
"a1" and "a2" successfully resulted in a 100% reduction of a
5.51-log E. coli inoculum after 180 minutes of contact in a large
scale setting without altering the color, texture and/or odor of
the treated produce.
EXAMPLE 4
[0178] Residual Composition After Drying and/or Washing
[0179] This Example demonstrates that residual amounts of the
invention's compositions remain on the treated surface, including
after rinsing off, to provide continued antimicrobial activity.
TABLE-US-00010 TABLE 9 Residual amounts in parts per million (ppm)
of component Component Spinach leaf Spring Mix Iceberg Lettuce
Propylene Glycol 25 13 18 Phosphoric Acid 8 9 5 Hydrogen Peroxide 0
0 0 Hypochlorous Acid 0.001 0.002 0.001 "CL2"
[0180] Table 9 shows the results of HPLC analysis of residual
amounts of components on 5 pounds of each of spinach leaf, spring
mix (containing baby lettuces), and iceberg lettuce following
treatment with the invention's compositions in accordance with
Table 8 above, And above. A flow through wash with 1 liter of
distilled water for 60 seconds was used for HPLC residual
tests.
EXAMPLE 5
[0181] Applying the Compositions Pre-Harvest
[0182] Prior to harvesting fruits or vegetables, compositions (A),
(B), (C) or (F) of Table 1 may be used as a pre-harvest treatment.
Early in the morning or late in the evening a harvester, tractor,
truck, ATV, utility vehicle or by hand could use a pre-mixed
composition in a tank used for spraying, fogging, sprinkling or
aerosoling. The fruit or vegetable would be covered by the
composition and allowed to dry to form a film. During this drying
time the antimicrobial action would continue. After the composition
has dried, the edible film left after the water has evaporated,
contains amounts of diol, acid and/or oxidizer that provide
continued antimicrobial activity and preservative functions.
EXAMPLE 6
[0183] Applying the Compositions During Harvesting
[0184] During harvesting of the agricultural products, compositions
(A), (B), (C), (D), (E), (F) or (G) of Table 1 may be used as a
harvesting treatment. A harvester, tractor, truck, ATV, utility
vehicle or by hand could use a pre-mixed composition in a tank used
for spraying, fogging, sprinkling or aero soling during picking
procedures. If a tow behind or driven harvester is used then a
dunking or immersion system may be implemented. Where the tubs,
crates, bins, boxes or container with fruits or vegetables may be
placed into a tank, basin or trough. This is done to thoroughly
cover the fruits or vegetables in the composition for a reasonable
amount of time to ensure complete contact. After the harvesting is
completed and the load of fruits or vegetables is shipped, the
composition dries to form a film. During this drying time the
antimicrobial activity would continue. After the composition has
dried, the edible film left after the water has evaporated,
contains amounts of diol, acid and/or oxidizer that provide
continued antimicrobial activity and preservative functions.
EXAMPLE 7
[0185] Applying the Compositions Before Washing and Bagging or
Crating
[0186] When fruits or vegetables are picked or harvested, they are
generally taken to a cold storage facility for chilling before they
are transported for additional processing or directly to food
services or stores. Before transporting, the fruits or vegetables
are generally packed into plastic containers that have openings
around all four sides for air circulation. The containers are
easily handled by forklift or other mechanical means. As the
forklift removes the containers and places them into a storage
(such as cold storage) area, the forklift could carry a tank with a
premixed composition of (A), (B), (C), (D), (E), (F) or (G) of
Table 1 and a spray apparatus to spray the composition on the
fruits or vegetables as the container is being moved. Once the
container has been moved to the cooling station facility, a
separate dipping or dousing station could be used to immerse the
container into composition (A), (B), (C), (D), (E), (F), (G), (H)
or (I) of Table 1 to cover the fruits or vegetables completely. The
immersion tank could use a chemical handling system that utilizes a
Microprocessor (analog or digital), pH electrode, Orp electrode,
Dissolved Oxygen electrode, Free Chlorine electrode or DPD system.
The system would use a signal representing the available
composition to turn on chemical pumps, valves or auger to add
additional chemicals and or water to make compositions for use in
the tank.
[0187] After the container has been immersed it would be removed to
drain and dry. During this drying time the antimicrobial activity
would continue. After the composition has dried, the edible film
left after the water has evaporated, contains amounts of diol, acid
and/or oxidizer that provide continued antimicrobial activity and
preservative functions.
EXAMPLE 8
[0188] Applying the Compositions During Washing and Bagging or
Crating.
[0189] Once the fruits or vegetables have been brought into the
washing and packaging facility they are typically conveyed either
by a belt, flume or motorized table. During this transit the fruits
or vegetables are washed with either aqueous sprays or aqueous
submersion. The composition (A), (B), (C), (D), (E), (F),(G), (H)
or (I) of Table 1 may be part of the aqueous solution used to clean
the fruits or vegetables. This solution may also contain various
oxidizers, defoamers, surfactants, oils, pesticides, dirt, and or
bugs. The composition could either mix into the added makeup water
or the recycled water. In either case the composition could be
inserted as combined ingredients or as independent ingredients. The
composition, either combined or independent, could be pump fed or
gravity fed into a suction side of a pump, tank inlet, vessel
inlet, trough inlet, flume inlet that is used for recycle or makeup
water, or into the main supply line for the sprays or for the
transport submersion. A microprocessor, analog or digital, pH
electrode, Orp electrode, Dissolved Oxygen electrode, Free Chlorine
electrode or DPD system may be used for automatic control. The
system would use a signal representing the available composition to
turn on chemical pumps, valves or auger to add additional
chemicals. Alternatively, a pre-mixture of the composition could be
added based on time, flow, level, turbidity, bio-load, microbial
load, etc.
[0190] After the fruits or vegetables have been washed they are
usually rinsed off. The composition could be in the rinse water.
After the fruits or vegetables are rinsed they are usually dried,
then metered to a packaging station. Alternately, prior to the
packaging station, the composition could be sprayed, fogged or
aerosoled, and the composition is allowed to dry into a very thin
film. The edible film left after the water has evaporated contains
amounts of diol, acid and/or oxidizer to provide continued
antimicrobial activity and preservative functions.
EXAMPLE 9
[0191] Shelf Life of Oxidizer.
[0192] This Example was carried out to determine the stability of
the exemplary oxidizer hypochlorous acid in the presence and
absence of diol and at different pH. The level of hypochlorous acid
was determined using a standard DPD assay.
TABLE-US-00011 TABLE 10 Hypochlorous acid stability and shelf life
Chlorine (ppm) Minutes Composition "x1a".sup.(a) Chlorine
Solution.sup.(b) 1 50 50 15 50 50 20 50 30 40 50 45 38 60 35 50 80
33 90 27 50 100 25 115 22 125 20 135 18 150 13 170 10 190 7 50 220
3 240 0 400 50 800 50 1600 48 2000 45 2300 43 2700 41 3000 40 3500
38 4300 36 .sup.(a)Composition "x1a" is referred to in Table 2, and
contains ortho-Phosphoric acid 0.03 wgt. %, propylene glycol 0.02
wgt. %, hypochlorous acid 0.005 wgt. % at pH 4 (FIG. 1A).
.sup.(b)"Chlorine solution" contains ortho-phosphoric acid 0.001
wgt. % and hypochlorous acid 0.005 wgt. % at pH 7 (FIG. 1B).
[0193] The data in Table 10 and FIG. 1 show that hypochlorous acid
in solution was less stable in the presence of diol at pH 4,
reaching 0% at about 4 hours, compared to 100% in the absence of
diol at pH 7.
[0194] Each and every publication and patent mentioned in the above
specification is herein incorporated by reference in its entirety
for all purposes. Various modifications and variations of the
described methods and system of the invention will be apparent to
those skilled in the art without departing from the scope and
spirit of the invention. Although the invention has been described
in connection with specific embodiments, the invention as claimed
should not be unduly limited to such specific embodiments. Indeed,
various modifications of the described modes for carrying out the
invention which are obvious to those skilled in the art and in
fields related thereto are intended to be within the scope of the
following claims.
* * * * *