Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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METHODS FOR INHIBITING MICROBIAL INFECTIONS ASSOCIATED WITH SANITARY PRODUCTS
CROSS REFERENCE TO RELATED APPLICATIONS
S This application is a continuation-in-part of copending International
Application Serial
No. PCT/US98/07307, filed April 10, 1998, which is a continuation-in-part of
United States
Provisional Application Serial No. 60/044,643, filed April 18, 1997, (now
abandoned), the
disclosures of which are hereby incorporated by reference herein.
TECHNICAL FIELD
The claimed invention relates to systems and methods to inhibit microbial
infections and
to promote epithelial probiosis when using sanitary health care products such
as disposable
diapers and other sanitary products. In particular, the claimed invention
describes use of
probiotic lactic acid bacteria in combination with sanitary health care
products to inhibit
microbial infections, promote dermal probiosis, and enhance biodegradatability
of disposed
sanitary products.
BACKGROUND OF THE INVENTION
Sanitary products are widely used in various formats for personal hygiene and
medical
necessity, and include sanitary napkins, diapers, incontinence guards, wound
dressings and the
like. By their use, a local tissue environment is produced which promotes
growth of microbial
pathogens, local infections, irritation, rashes, and related problems.
In addition, disposal of used sanitary products is a major environmental and
health care
concern. The volume of material and the type of material present in the used
sanitary product,
due to its absorbent character and purpose of collecting body fluids and waste
materials, provides
a biological and environmental hazard in disposal of used sanitary products.
There is a great
need for improvements in biodegradation of used sanitary products -
degradation of both the
product itself and the waste product it contains.
The claimed invention uses a bacterium that is probiotic and heterotrophic to
resolve both
of the above problems (i.e., to inhibit microbial infections associated with
use of sanitary
products, and to promote biodegradation of the sanitary product after use).
Probiotic microorganisms are those which confer a benefit when grow in a
particular
environment, often by inhibiting the growth of other biological organisms in
the same
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environment. Examples of probiotic organisms include bacteria and
bacteriophages which
possess the ability to grow within the gastrointestinal tract, at least
temporarily, to displace or
destroy pathogenic organisms, as well as providing other benefits to the host.
See e.g., Salminen
et al, 1996. Antonie Yan Leeuwenhoek 70: 347-358; Elmer et al, 1996. JAMA 275:
870-876;
Rafter, 1995. Scand. J. Gastroenterol. 30: 497-502; Perdigon et al, 1995. J.
Dairy Sci. 78: 1597-
1606; Gandi, Townsend Lett. Doctors & Patients, pp. 108-110, Jan. 1994;
Lidbeck et al, 1992.
Eur. J. Cancer Prev. l: 341-353.
The majority of previous studies on probiosis have been observational rather
than
mechanistic in nature, and thus the processes responsible for many probiotic
phenomena have yet
to be quantitatively elucidated. Some probiotics are members of the normal
colonic microflora
and are not viewed as being overtly pathogenic. However, these organisms have
occasionally
caused infections (e.g., bacteremia) in individuals who are, for example,
immunocompromised.
See e.g., Sussman, J. et al., 1986. Rev Infect. Dis. 8: 771-776; Hata, D. et
al., 1988. Pediatr.
Infect. Dis. 7: 669-671.
1 S For example, the probiotic bacteria found in sour milk, has been utilized
since ancient
times (i.e., long-before the discovery of bacteria) as a therapeutic treatment
for dysentery and
related gastrointestinal diseases. More recently, probiotic preparations were
systematically
evaluated for their effect on health and longevity in the early-1900's (see
e.g., Metchinikoff, E.,
Prolongation of Life, Willaim Heinermann, London 1910), although their
utilization has been
markedly limited since the advent of antibiotics in the 1950's to treat
pathological microbes. See
e.g., Winberg, et al, 1993. Pediatr. Nephrol. 7: 509-514; Malin et al, Ann.
Nutr. Metab. 40: 137-
145; and U.S. Patent No. 5,176,911. Similarly, lactic acid-producing bacteria
(e.g., Bacillus,
Lactobacillus and Streptococcus species) have been utilized as food additives
and there have
been some claims that they provide nutritional and/or therapeutic value. See
e.g., Gorbach,
1990. Ann. Med. 22: 37-41; Reid et al, 1990. Clin. Microbiol. Rev. 3: 335-344.
The nutritional use of probiotic bacteria, especially Lactobacillus and
Biffidobacterium
strains, that colonize the gut has been previously disclosed (Winberg, et al.,
Pediatr. Nephrol. 7:
509-S 14, 1993; Malin, et al., Ann. Nutr. Metab. 40:137-145, 1996; and U.S.
Patent. No.
4,176,911). Lactic acid producing bacteria (e.g., Bacillus, Lactobacillus and
Streptococcus
species) have been used as food additives and there have been some claims that
they provide
nutritional and therapeutic value (Gorbach, Ann. Med. 22: 37-41, 1990; Reid,
et al., Clin.
Microbiol. Rev., 3: 335-344, 1990). Heterotrophic bacteria play an important
role in the
biodegradation of animal waste and many natural and synthetic polymers.
Bacterial strains
including: Bacillus, Pseudomonas, Arthrobacter, Achromobacter, Micrococcus and
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Rhodococcus have been shown to participate in the breakdown of waste products,
cellulose
materials, petroleum hydrocarbons and their associated products, such as
plastics, synthetic
rubbers and other synthetic materials.
Bacillus coagulans is a non-pathogenic gram positive spore-forming bacteria
that
produces L(+) lactic acid (dextrorotatory) in homofermentation conditions. It
has been isolated
from natural sources, such as heat-treated soil samples inoculated into
nutrient medium
(Bergey's Manual off Systemic Bacteriology, Vol. 2, Sneath, P.H.A., et al.,
eds., Williams &
Wilkins, Baltimore, MD, 1986). Purified B. coagulans strains have served as a
source of
enzymes including endonucleases (e.g., U.S. Patent No. 5,200,336); amylase
(U.S. Patent No.
4,980,180); lactase (U.S. Patent No. 4,323,651); and cyclo-malto-dextrin
glucano-transferase
(U.S. Patent No. 5,102,800). B. coagulans has been used to produce lactic acid
(U.S. Patent No.
5,079,164). A strain of B. coagulans (referred to as L. sporogenes; Sakaguti &
Nakayama
(ATCC 31284)) has been combined with other lactic acid producing bacteria and
B. natto to
produce a fermented food product from steamed soybeans (U.S. Patent No.
4,110,477).
Use of a sanitary product produces frequent dermal mucoidal irntations and/or
infections
associated with the use of the product. Diaper rash is a common issue in both
adults and infants.
Rashes can become more serious irntations when opportunistic pathogens
introduced into the
sanitary product germinate and cause infections on these irntated sites. In
addition, vulva-
vaginal infections are common with the use of napkins and tampons and are
typically caused by
Candida or Gardnerella species (e.g., Candida albicans and C. tropicalis).
Toxic Shock
Syndrome and other dermal infections caused by Staphylococcal bacteria (e.g.,
Staphylococcus
aureus and S. epidermidis are also common). Other pathogens which can cause
infection after
brief periods of dermal irritation and/or use of sanitary products include
Trichophyton species
(e.g., T. mentagrophytes).
In addition, disposable diapers and other sanitary products present
environmental
problems in their disposal. Sanitary landfills are overused and accumulate
excessive amounts of
disposed products. Sanitary products such as diapers, sanitary napkins and
tampons biodegrade
slowly and occupy considerable space due to the bulk of these products,
particularly when
containing body excrements or fluids which expand due to their absorbent
polymer content.
SUMMARY OF THE INVENTION
It has now been discovered that probiotic acid-producing bacteria are
effective in
inhibiting, preventing and/or eliminating dermal/epithelial infections by
preventing the growth of
dermal pathogens which grow upon use of diapers and other sanitary products.
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It has also been discovered that bacterial enzymes and other metabolic
products of
probiotic acid-producing bacteria play an important role in the biodegradation
of many sanitary
products, including biodegradation of the waste biomaterials, such as when
disposed in landfills.
The claimed invention discloses compositions and articles of manufacture
containing
non-pathogenic probiotic acid-producing bacteria, and their methods of use for
inhibiting
pathogen growth on skin in applications where sanitary products are used,
which also provides
for degradation of the sanitary products and body waste products contained
thereby. The
invention contemplates sanitary products as articles of manufacture which
contain effective
amounts of a probiotic bacterium in various parts of the product so as to
achieve the desired
result of inhibiting microbial infections on the tissues in contact with the
sanitary product and/or
enhance biodegradation of the sanitary product and waste products collected
upon the sanitary
product.
Typically, the probiotic acid-producing bacteria is introduced into or onto
portions of the
sanitary product by applying a composition containing viable bacteria to the
product during a
stage of the manufacture of the sanitary product.
In preferred embodiments, the invention contemplates using a lactic acid-
producing
bacteria, and more preferably using spore-forming Bacillus species,
particularly B. coagulans,
being a preferred embodiment, and B. coagulans Hammer being a particularly
preferred
embodiment.
In one embodiment of the composition, a Bacillus coagulans strain is included
in the
composition in the form of spores. In another embodiment, a Bacillus coagulans
strain is
included in the composition in the form of a dried cell mass. In another
embodiment, a Bacillus
coagulans strain is included in the composition in the form of a stabilized
paste. In another
embodiment, a Bacillus coagulans strain is included in the composition in the
form of stabilized
gel.
In one embodiment, the composition further includes an effective amount of a
bifidogenic oligosaccharide, such as a short or long chain
fructooligosaccharide (FOS), a
gluco-oligosaccharide or other long-chain oligosaccharide polymer not readily
digested by
pathogenic bacteria as described herein.
It should be understood that both the foregoing general description and the
following
detailed description are exemplary and explanatory only and are not
restrictive of the invention
as claimed.
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DETAILED DESCRIPTION OF THE INVENTION
The present invention is directed to the discovery that non-pathogenic lactic
acid-
producing bacteria (i.e., "lactic acid bacteria"), such as the exemplary
Bacillus coagulans, can be
used in compositions as a probiotic for inhibiting growth of microbial dermal
and epithelial
pathogens which can occur upon use of various sanitary products. Typically,
these bacteria can
be used as a preventative or ameliorative treatment of rashes and other dermal
infections that
manifest themselves as a result of use of the sanitary product, including
irritated/inflamed skin,
dermatitis, excema, or skin allergies.
In addition, the invention describes the use of one or more bacteria species
to enhance
degradation of the sanitary product, which are included in the compositions,
methods and articles
of manufacture of this invention and are referred to herein as "degradative
bacteria".
A sanitary product can be any of variety of materials used in contact with a
body tissue
which, upon use, is susceptible to dermal or epithelial rashes and/or
infections. Exemplary
sanitary products include an infant or adult diaper, sanitary napkin, tampon,
incontinence guard,
bed sheet or protector, wound or sore dressing, dermal patch, adhesive tape,
saliva absorbent,
and related disposable sanitary and hygiene products, although the invention
need not be viewed
as so limited.
The invention therefore describes various compositions, methods for using the
compositions and systems containing the compositions. In preferred
embodiments, the
composition further comprises an effective amount of a bifidogenic
oligosaccharide, such as
fructo-oligosaccharide (FOS), gluco-oligosaccharide (GOS) and the like, as
described herein.
A. Probiotic Acid-Producing Bacteria
A probiotic acid-producing bacteria suitable for use in the methods and
compositions of
the invention as defined for use in the present invention produces acid and is
non-pathogenic.
There are many suitable bacteria identified as described herein, although the
invention is not
limited to currently known bacterial species insofar as the purposes and
objectives of the bacteria
is described. The property of acid production is key to the effectiveness of
the probiotic lactic
acid-producing bacteria of this invention because the lactic acid production
increases acidity in
the local microfloral environment, which does not support growth of many
deleterious and
undesirable bacteria and fungi. By the mechanism of lactic acid production,
the probiotic
inhibits growth of competing and deleterious bacteria.
As used herein, "probiotic" refers to microorganisms that form at least a part
of the
transient or endogenous flora and thereby exhibit a beneficial prophylactic
and/or therapeutic
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effect on the host organism. Probiotics are generally known to be safe by
those skilled in the art
(i. e., non-pathogenic). Although not wishing to be bound by any particular
mechanism, the
prophylactic and/or therapeutic effect of an lactic acid-producing bacteria of
this invention
results in part from competitive inhibition of growth of pathogens due to
superior colonization,
parasitism of undesirable microorganisms, lactic acid production and/or other
extracellular
products having antimicrobial activity, or combinations thereof. These
products and activities of
an lactic acid-producing bacteria of this invention act synergistically to
produce the beneficial
probiotic effect.
Typical lactic acid-producing bacteria (i.e., a " lactic acid bacteria")
useful as a probiotic
of this invention are efficient acid producers which include non-pathogenic
members of the
Bacillus genus, all members of the Lactobacillus and Sporolactobacillus genus,
all members of
the Bifidobacterium genus, and Pseudomonas limbergii, although certain species
are particularly
preferred as described herein.
Preferred lactic acid-producing bacteria include the Bacillus laterosporus or
Bacillus
subtilis species described herein, including Bacillus laterosporus, Bacillus
laterosporus BOD,
Bacillus laterosporus laubach and Bacillus subtilis.
More preferably, the present invention contemplates the use of a lactic acid-
producing
bacteria ("lactic acid bacteria") which includes the above Lactobacillus,
Bifidobacterium and
certain Bacillus species. Particularly preferred are lactic acid-producing
bacteria, such as
L. sporogenes (aka B. coagulans), Sporolactobacillus P44, and Bacillus brevis
subsp. coagulans.
Exemplary lactic acid-producing, non-pathogenic Bacillus species are Bacillus
coagulans, Bacillus coagulans Hammer, Bacillus brevis subspecies coagulans and
Bacillus
laevolacticus.
Exemplary lactic acid-producing Lactobacillus species include Lactobacillus
acidophilus,
Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus cereale,
Lactobacillus DDS-l,
Lactobacillus delbrukeii, Lactobacillus fermentum, Lactobacillus gaserii,
Lactobacillus GG,
Lactobacillus jensenii, Lactobacillus rhamnosus, Lactobacillus plantarum,
Lactobacillus
salivarius, Lactobacillus sporogenes (aka B. coagulans) and Lactobacillus
thermophilus.
Exemplary lactic acid-producing Sporolactobacillus species include all
Sporolactobacillus species, including Sporolactobacillus P44.
Exemplary lactic acid-producing Bifidobacterium species include:
Bifidobacterium
adolescentis, Bifidobacterium animalis, Bifidobacterium bifidum,
Bifidobacterium bifidus,
Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium infantus and
Bifidobacterium
longum.
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There are several Bacillus species particularly useful as a probiotic
according to the
present invention, including the lactic acid-producers Bacillus coagulans, and
Bacillus
laevolacticus.
It should be noted that although exemplary of the present invention, Bacillus
coagulans is
only a model for the other lactic acid producing species of probiotic bacteria
useful in the
practice of the present invention, and therefore the invention is not to be
considered as limiting
and it is intended that any of the acid producing species of probiotic
bacteria can be used in the
compositions, therapeutic systems and methods of the present invention.
There are a variety of different Bacillus species useful in the present
invention, including,
but not limited to many different strains available through commercial and
public sources, such
as the American Type Culture Collection (ATCC). For example, Bacillus
coagulans strains are
available as ATCC Accession Numbers 15949, 8038, 35670, 11369, 23498, 51232,
11014,
31284, 12245, 10545 and 7050. Bacillus laevolacticus strains are available as
ATCC Accession
Numbers 23495, 23493, 23494, 23549 and 23492. A Bacillus species is
particularly suited for
1 S the present invention, particularly species having the ability to form
spores which are relatively
resistant to heat and other conditions, making them ideal for storage (shelf
life) in product
formulations, and ideal for survival and colonization of tissues under
conditions of pH, salinity,
and the like on tissues of the skin and epithelium. Additional useful
properties include being
non-pathogenic, aerobic, facultative and heterotrophic, rendering these
species safe, and able to
colonize skin and epithelium.
Because Bacillus spores are heat-resistant and additionally can be stored as a
dry power,
they are particularly useful for formulation into and manufacture of dry
products such as the
various sanitary products and compositions described herein. Heat and pressure-
resistant spores
are also suitable for use in pressure-treated absorbent compositions described
herein.
Exemplary methods and compositions are described herein using Bacillus
coagulans as a
probiotic. Purified Bacillus coagulans is particularly useful as a probiotic
in the present
invention. Probiotic B. coagulans is non-pathogenic and is generally regarded
as safe (i.e.,
GRAS classification) by the U.S. Federal Drug Administration (FDA) and the
U.S. Department
of Agriculture (USDA), and by those skilled in the art. The Gram positive rods
of B. coagulans
have a cell diameter of greater than 1.0 micrometer ( m ) with variable
swelling of the
sporangium, without parasporal crystal production.
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Because B. coagulans forms heat-resistant spores, it is particularly useful
for making
pharmaceutical compositions' that require heat and pressure in their
manufacture. Formulations
that include viable B. coagulans spores in a pharmaceutically acceptable,
carrier are particularly
preferred for making and using compositions according to the present
invention.
The growth of these various Bacillus species to form cell cultures, cell
pastes and spore
preparations is generally well known in the art. Exemplary culture and
preparative methods are
described herein for Bacillus coagulans and can readily be used and/or
modified for growth of
the other lactic acid producing bacteria of this invention.
1. Sources of Bacillus coagulans
Purified Bacillus coagulans bacteria utilized in the present invention are
available from
the American Type Culture Collection (ATCC, Rockville, MD) using the following
accession
numbers: Bacillus coagulans Hammer NRS 727 (ATCC No. 11014); Bacillus
coagulans
Hammer strain C (ATCC No. 11369); Bacillus coagulans Hammer (ATCC No. 31284);
and
Bacillus coagulans Hammer NCA 4259 (ATCC No. 15949). Purified Bacillus
coagulans
bacteria are also available from the Deutsche Sarumlung von Mikroorganismen
and Zellkuturen
GmbH (Braunschweig, Germany) using the following accession numbers: Bacillus
coagulans
Hammer 1915 (DSM No. 2356); Bacillus coagulans Hammer 1915 (DSM No. 2383,
corresponds to ATCC No. 11014); Bacillus coagulans Hammer (DSM No. 2384,
corresponds to
ATCC No. 11369); and Bacillus coagulans Hammer (DSM No. 2385, corresponds to
ATCC No.
15949). Bacillus coagulans bacteria can also be obtained from commercial
suppliers such as
Sabinsa Corporation (Piscataway, NJ) or K.K. Fermentation (Kyoto, Japan).
Bacillus coagulans strains and their growth requirements have been described
previously
(see e.g., Baker, D. et al, 1960. Can. J. Microbiol. 6: 557-563; Nakamura, H.
et al, 1988. Int. J.
Svst. Bacteriol. 38: 63-73. In addition, various strains of Bacillus coagulans
can also be isolated
from natural sources (e.g., heat-treated soil samples) using well-known
procedures (see e.g.,
Bergey's Manual of Systemic Bacteriology, Vol. 2, p. 1117, Sneath, P.H.A. et
al., eds., Williams
& Wilkins, Baltimore, MD, 1986).
It should be noted that Bacillus coagulans had previously been mis-
characterized as a
Lactobacillus in view of the fact that, as originally described, this
bacterium was labeled as
Lactobacillus sporogenes (Nakamura, et al., Int. J. Syst. Bacteriol. 38: 63-
73, 1988). However,
initial classification was incorrect due to the fact that Bacillus coagulans
produces spores and
through metabolism excretes L(+)-lactic acid, both aspects which provide key
features to its
utility. Instead, these developmental and metabolic aspects required that the
bacterium be
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classified as a lactic acid bacillus, and therefore it was re-designated. In
addition, it is not
generally appreciated that classic Lactobacillus species are unsuitable for
colonization of the gut
due to their instability in the harsh (i.e., acidic) pH environment of the
bile, particularly human
bile. In contrast, Bacillus coagulans is able to survive and colonize the
gastrointestinal tract in
the bile environment and even grown in this low pH range. In particular, the
human bile
environment is different from the bile environment of animal models, and
heretofore there has
not been any accurate descriptions of Bacillus coagulans growth in human
gastrointestinal tract
models.
2. Growth of Bacillus coagulans
Bacillus coagulans is aerobic and facultative, grown typically in nutrient
broth, pH 5.7 to
6.8, containing up to 2% (by wt) NaCI, although neither NaCI nor KCl are
required for growth.
A pH of about 4 to about 7.5 is optimum for initiation of growth from spores.
It is optimally
grown at about 30°C to about 45°C, and the spores can withstand
pasteurization. It exhibits
facultative and heterotrophic growth by utilizing a nitrate or sulfate source.
Additional metabolic
characteristics of B. coagulans are summarized in Table 1.
TABLE 1
Characteristic Bacillus coa~ulahs Res onse
Catalase production Yes
Acid from D-Glucose Yes
Acid from L-Arabinose Variable
Acid from D-Xylose Variable
Acid from D-Mannitol Variable
Gas from Glucose Yes
Hydrolysis of Casein Variable
Hydrolysis of Gelatin No
Hydrolysis of Starch Yes
Utilization of Citrate Variable
Utilization of Propionate No
Degradation of Tyrosine No
Degradation of Phenylalanine No
Nitrate reduced to Nitrite Variable
Allatoin or Urate Required No
Bacillus coagulans can be grown in a variety of media, although it has been
found that
certain growth conditions produce a culture which yields a high level of
sporulation. For
example, sporulation is enhanced if the culture medium includes 10 milligrams
per liter of
manganese sulfate, yielding a ratio of spores to vegetative cells of about
80:20. In addition,
certain growth conditions produce a bacterial spore which contains a spectrum
of metabolic
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enzymes particularly suited for the present invention (i.e., production of
lactic acid and enzymes
for the enhanced probiotic activity and biodegradation). Although spores
produced by these
particular growth conditions are preferred, spores produced by any compatible
growth conditions
are suitable for producing a B. coagulans useful in the present invention.
Suitable media for growth of B. coagulans include Nutristart 701, 2DB (potato
dextrose
broth), TSB (tryptic soy broth) and NB (nutrient broth),~all well known and
available from a
variety of sources. Media supplements containing enzymatic digests of poultry
and fish tissue,
and containing food yeast are particularly preferred. A preferred supplement
produces a media
containing at least 60% protein, and about 20% complex carbohydrates and 6%
lipids. Media
can be obtained from a variety of commercial sources, notably DIFCO (Detroit,
MI), Oxoid
(Newark, NJ), BEL (Cockeyesville, MD), Advanced Microbial Systems, (Shakopee,
MN), and
Tray Biologicals (Tray, MI) A preferred procedure for preparation of B.
coagulans is described
in the Examples section, infra.
3. Probiotic Antimicrobial Activity of Bacillus coagulans
1 S One aspect of the utility of B. coagulans in the present invention is
based on the ability of
probiotic B. coagulans to inhibit growth of pathogenic enteric microorganisms
as described in
the Examples. Pathogenic bacteria inhibited by B. coagulans activity include:
Staphylococcus
aureus, S. epidermidis, Streptococcus pyogenes, Pseudomonas aeruginosa,
Escherichia coli
(enterohemorragic species), Clostridium species including C. perfingens, C.
difficile,
C. botulinum, C. tributrycum, and C. sporogenes, Gardnerella vaginalis,
Propionibacterium
acnes, Aeromonas hydrophilia, Candida species, Proteus species, Klebsiella
species, fungal
dermatophytes or other mycotic pathogens. These pathogens can cause a variety
of pathologies
including disruption of normal tissue function, and the like conditions as are
well known in the
art. Therefore, use of compositions containing a probiotic that inhibits these
pathogens are
useful in preventing or treating conditions associated with infection by these
pathogens.
Although B. coagulans is exemplary, by virtue of the common properties of a
lactic acid-
producing bacteria, a therapeutic composition comprising an acid bacterium of
this invention can
be used in a method or composition of this invention.
4. Extracellular Products Having Antimicrobial Activity
B. coagulans cultures contain secreted products which have antimicrobial
activity. These
secreted products are useful in therapeutic compositions according to the
present invention. Cell
cultures are harvested as described herein, and the culture supernatants are
collected, by filtration
or centrifugation, or both, and the resulting supernatant contains
antimicrobial activity useful in a
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therapeutic composition. The preparation of a B. coagulans extracellular
product is described in
the Examples section, infra.
Extracellular products of B. coagulans may be included in compositions for use
in the
invention. In particular, an effective amount of an extracellular product can
be applied to a
structural component part of a sanitary product of this invention, such as a
diaper, bandage,
sanitary napkin, tampon and the like product.
B. Degradative Bacteria
In one embodiment, the claimed invention contemplates that the bacteria used
in a
sanitary product, system or related method according to the present invention
have the ability to
support biodegradation of the product as an additional feature of the
invention. To that end the
product, system or method comprises a degradative bacteria as described
herein.
A degradative bacteria, or "degradation-enhancing" non-pathogenic bacteria,
can be any
of the bacteria previously recited which are defined as a probiotic, lactic
acid- producing bacteria
herein which have the property of being a degradative bacteria, or can be a
different bacteria,
such that two different species of bacteria are used in practicing the claimed
invention. That is,
is one embodiment both a probiotic acid-producing bacteria and a degradative
bacteria are
included in a composition, product, system or method according to the present
invention.
Typical bacterial are any non-pathogenic bacteria which promotes degradation
of human
waste products, and preferably also can degrade the absorbent materials of the
sanitary product
of the present invention. A preferred bacteria is any non-pathogenic member of
the Bacillus
genus, Lactobacillus genus, Sporolactobacillus genus, Bifidobacterium genus,
Pseudomonas
genus, and the like bacteria.
Particularly preferred members of the Bacillus genus include: Bacillus
acidocaldarius,
Bacillus alcalophilus, Bacillus azotoformans, Bacillus badius, Bacillus
brevis, Bacillus brevis
subsp. coagulans, Bacillus cereus, Bacillus chitinosporus, Bacillus circulans,
Bacillus
coagulans, Bacillus dextrolacticus, Bacillus firmus, Bacillus globisporus,
Bacillus hydrophilus,
Bacillus laevolacticus, Bacillus laterosporus, Bacillus laterosporus BOD,
Bacillus laterosporus
Laubach, Bacillus lentus, Bacillus licheniformis, Bacillus macerans, Bacillus
marinus, Bacillus
megaterium, Bacillus modestus, Bacillus mycoides, Bacillus pantothenticus,
Bacillus pumilus,
Bacillus polymyxa, Bacillus smithii, Bacillus stereothermophilus, Bacillus
subtilis, Bacillus
thermoacidurans, Bacillus thuringiensis, Bacillus uniflagellatus, and the
like.
Particularly preferred members of the Pseudomonas genus include Pseudomonas
alcaligenes, Pseudomonas limbergii, Pseudomonas pseudoalcaligenes, and
Pseudomonas 679-2.
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Particularly preferred members of the Bifidobacterium genus include
Bifidobacterium
adolescentis, Bifidobacterium animalis, Bifidobacterium bifidum,
Bifidobacterium bifidus,
Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium infantus,
Bifidobacterium
longum, and the like.
Particularly preferred members of the Lactobacillus genus include
Lactobacillus
acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus
cereale, Lactobacillus
delbrukeii, Lactobacillus DDS l, Lactobacillus fermentum, Lactobacillus
gaserii, Lactobacillus
GG, Lactobacillus jensenii, Lactobacillus plantarum, Lactobacillus rhamnosus,
Lactobacillus
salivarius, Lactobacillus sporogenes, Lactobacillus thermophilus, and the
like.
A preferred member of the Sporolactobacillus genus include Sporolactobacillus
P44.
Degradation of waste products or sanitary products can be readily assessed by
any of a
variety of processes, and therefore the term "degradation-enhancing" is not to
be construed as so
limited to any particular degree or rate of enhancement of degradation.
The above bacteria are well known in the bacterial arts, and can be obtained
from well
known sources and propagated by well known methods. An exemplary source is the
American
Type Culture Collection (ATCC), although other culture banks are also
available. The culturing
of bacteria is also well known, and particularly the preparation of spores for
the sporulating
varieties of bacteria, which are particularly preferred.
C. Bifidogenic Oligosaccharides
Bifidogenic oligosaccharides, as used in the context of the present invention,
are a class
of sugars particularly useful for preferentially promoting the growth of a
lactic acid bacteria of
this invention. These oligosaccharides include fructo-oligosaccharides (FOS),
gluco-
oligosaccharides (GOS), other long-chain oligosaccharide polymers of fructose
and/or glucose,
and the trisaccharide raffinose, all of which are not readily digested by
pathogenic bacteria. The
preferential growth is promoted due to the nutrient requirements of this class
of lactic acid
bacterium as compared to pathogenic bacteria. Bifidogenic oligosaccharides are
polymers that
are utilized almost exclusively by the indigenous Bifidobacteria and
Lactobacillus and can be
similarly utilized by Bacillus. Deleterious microorganisms such as
Clostridium, Candida,
Campylobacter, Klebsiella, Pseudomonas, Staphylococcus, Salmonella and E. coli
cannot
metabolize FOS or other bifidogenic oligosaccharides, and therefor use of
these bifidogenic
oligosaccharides in combination with a lactic acid bacteria of this invention,
particularly
Bacillus, allows the beneficial and probiotic bacteria to grow and to replace
any undesirable or
pathogenic microorganisms.
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The use of bifidogenic oligosaccharides in compositions of the present
invention provides
a synergistic effect thereby increasing the effectiveness of the probiotic-
containing compositions
of this invention. This synergy is manifest at least by selectively increasing
the ability of the
probiotic bacterium to grow by increasing the food supplement for probiotic
bacteria which
preferentially selects for growth of the probiotic bacteria over many other
bacterial species in the
infected tissue. In addition, it is understood that Bifidobacteria and
Lactobacillus are also
producers of lactic acid. Bifidogenic oligosaccharides enable these probiotic
organisms to
proliferate preferentially over the undesirable bacteria that may be present
in the tissue to be
treated by the present invention, thereby furthering the probiotic state of
the body. Thus, the
presence of the bifidogenic oligosaccharides in the formulation allows for
more effective
inhibition of undesirable microbes by increasing the ability of all varieties
of beneficial probiotic
bacteria to grow and therefore provide benefit.
The bifidogenic oligosaccharide can be used either alone or in combination
with a lactic
acid bacterium in a composition. That is, due to the growth promoting activity
of bifidogenic
oligosaccharides, the invention also contemplates a composition comprising a
bifidogenic
oligosaccharide of this invention in a lactic acid bacterium growth-promoting
amount. As shown
herein, these amounts can vary widely since a probiotic lactic acid bacterium
will respond to any
metabolic amount of nutrient oligosaccharide, and therefore the invention need
not be so limited.
A preferred and exemplary bifidogenic oligosaccharide is FOS, although the
other sugars
can also be utilized, either alone or in combination. FOS can be obtained from
a variety of
natural sources, including commercial suppliers. As a product isolated from
natural sources, the
components can vary widely and still provide the beneficial agent, namely FOS.
FOS typically
has a polymer chain length of from about 4 to 100 sugar units, with the longer
lengths being
preferred. For example, the degree of purity can vary widely so long as
functional FOS is
present in the formulation. Preferred FOS formulations contain at least 50% by
weight of fructo-
oligosaccharides compared to simple (mono or disaccharide) sugars such as
glucose, fructose or
sucrose, preferably at least 30% fructo-oligosaccharides, more preferably at
least 90% and most
preferably at least 95% fructo-oligosaccharides. Sugar content and composition
can be
determined by any of a variety of complex carbohydrate analytical detection
methods as is well
known.
Preferred sources of FOS include: inulin, Frutafit IQTM from Imperial Suiker
Unie (Sugar
Land, Texas), NutraFIoraTM from Americal Ingredients, Inc., (Anaheim, CA),
Fabrchem, Inc.,
(Fairfield, CT), and Fruittrimfat Replacers and Sweeteners (Emeryville, CA).
Bifidogenic
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oligosaccharides such as GOS, and other long chain oligosaccharides are also
available from
commercial vendors.
D. Compositions
The present invention is directed to the discovery that lactic acid bacteria,
particularly
Bacillus species, can be used in compositions as a probiotic in combination
with sanitary
products for inhibiting dermal and epithelial microbial infections associated
with the use of
sanitary products. As discussed further, the compositions can be formulated in
many
configurations because the bacterium can be presented as a viable organism
(e.g., as a vegetative
cell or as a spore depending on the species and form of probiotic organism)
and colonize tissues
associated with use of a sanitary product. The cells/spores can be presented
in a variety of
compositions suited for use in a sanitary product.
The active ingredients (i.e., live bacteria or extracellular components,
comprise about
0.1 % to about 50% by weight of the final composition, preferably 1 % to 10%
by weight in the
composition. A typical composition will contain in a one gram dosage
formulation a
concentration of from 103 to 10'4 colony forming units (CFU) of viable acid
bacterium (i.e.,
vegetative cell) or bacterial spore, preferably 105 to 10' 2 colony forming
units/g, whereas in other
preferred embodiments the concentrations are 109 to 10'3 colony forming
units/g; 105 to 10'
colony forming units/g; or 108 to 109 colony forming units/g.
In one embodiment, a composition for use in a sanitary product according to
the present
invention can further comprise a degradation-enhancing bacteria as described
herein. A preferred
amount of this bacteria is an amount sufficient to promote degradation, which
can be from about
104 to 10'4 CFU of viable bacteria for use per unit of sanitary product,
preferably about 10' to
10'° CFU per unit, and more preferably about 108 to 109 CFU per unit.
The actual amount in a
composition will vary depending upon the amounts of composition to be
dispersed into the
absorbent structure or other portions of the sanitary product, and upon routes
of dispersal.
In addition, it is contemplated that combinations of bacteria may be utilized
to afford
optimized formulations depending upon the circumstances. Thus, various
combinations of
species of bacteria may be used, and in varying amounts, so long as the
primary objective is to
provide a probiotic acid bacteria, and the secondary objective is to provide a
degradation-
enhancing bacteria.
In a preferred embodiment, the invention contemplates certain preferred
combinations.
In particular, a mixture of bacterial spores is ideally suited for
manufacturing and shelf storage.
A preferred embodiment utilizes a dispersion of about one billion CFU of B.
coagulans spores
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mixed with about one billion CFU of degradation enhancing bacterial spores,
preferably an equal
mix of B. licheniformis, B. subtilis, B. pumilis and B. megaterium.
In one preferred embodiment a composition may further include from about 10
milligrams (mg) to one gram of a bifidogenic oligosaccharide, preferably a
fructo-
oligosaccharide. The composition typically contains a lactic acid bacterium
growth-promoting
amount of the bifidogenic oligosaccharide, which growth-promoting amount can
vary widely and
be readily measured by growth assays as described herein. The composition will
typically
contain 10 mg to 1 gm of bifidogenic oligosaccharide per gram of composition
depending on the
dosage, route of administration and intended usage.
The formulation for a composition of this invention may include other
probiotic agents or
nutrients for promoting spore germination and/or bacterial growth. A
particularly preferred
material is a bifidogenic factor which promotes growth of beneficial probiotic
bacteria as
described herein.
The compositions may also include known antimicrobial agents, known antiviral
agents,
known antifungal agents, all of which must be compatible with maintaining
viability of the
Bacillus active agent when Bacillus organisms or spores are the active agent.
The other agents in
the compositions can be either synergists or active agents. Preferably, the
known antimicrobial,
antiviral and/or antifungal agents are probiotic agents compatible with
Bacillus. The
compositions may also include known antioxidants, buffering agents, and other
agents such as
coloring agents, flavorings, vitamins or minerals. Thickening agents may be
added to the
compositions such as polyvinylpyrrolidone, polyethylene glycol or
carboxymethylcellulose.
Chemicals used in the present compositions can be obtained from a variety of
commercial sources, including Spectrum Quality Products, Inc (Gardena, CA),
Seltzer
Chemicals, Inc., (Carlsbad, CA) and Jarchem Industries, Inc., (Newark, NJ).
The active agents are combined with a carrier that is physiologically
compatible with the
dermal or epithelial tissue of a human or animal to which it is administered.
That is, the carrier
is preferably substantially inactive except for surfactant properties used in
making a suspension
of the active ingredients. The compositions may include other physiologically
active
constituents that do not interfere with the efficacy of the active agents in
the composition.
A formulated composition of this invention may be completed in weight using
any of a
variety of carriers and/or binders. A preferred carrier is micro-crystalline
cellulose (MCC) added
in an amount sufficient to complete the one gram dosage total weight.
Particularly preferred
formulations for a composition of this invention are described in the Examples
section, infra.
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Carriers can be solid-based dry materials for formulations in tablet, granule
or powdered
form, and can be liquid or gel-based materials for formulations in liquid or
gel forms, which
forms depend, in part, upon the manner of use or the manner of manufacturing a
sanitary
product. Typical carriers for dry formulations include trehalose, malto-
dextrin, rice flour, micro-
s crystalline cellulose (MCC) magnesium stearate, inositol, FOS, gluco-
oligosaccharides (GOS),
dextrose, sucrose, and the like carriers.
Where the composition is dry and includes evaporated oils that produce a
tendency for
the composition to cake (adherence of the component spores, salts, powders and
oils), it is
preferred to include dry fillers which distribute the components and prevent
caking. Exemplary
anti-caking agents include MCC, talc, diatomaceous earth, amorphous silica and
the like,
typically added in an amount of from about 1 to 95 % by weight.
The carrier is preferably a formulation in which, for example, B. coagulans
can be
suspended for hydration by the user before it is administered to the sanitary
product or tissue.
Suitable liquid or gel-based Garners are well known in the art, such as water
and physiological
salt solutions, urea, alcohols and glycols such as methanol, ethanol,
propanol, butanol, ethylene
glycol and propylene glycol, and the like. Preferably, water-based Garners are
about neutral pH.
In a related embodiment, the invention describes an aqueous liquid absorbent
composition comprising an aqueous liquid absorbing medium (i.e., an
"absorbent") and a
microbe-inhibiting amount of an extracellular product isolated from B.
coagulans as described
herein. he absorbent composition is similar to an absorbent structure
described herein for a
sanitary product insofar as the composition in the absorbent portion of the
sanitary product. The
composition is formulated to be added to or dispersed into/onto a sanitary
product for use of the
product according to the methods described for a sanitary product of the
present invention, that is
to inhibit microbial growth upon use of the product.
The composition containing absorbent and the extracellular product is
typically produced
by admixing the extracellular product with a pre-selected amount of absorbent,
and drying or
desiccating the admixture to coat the absorbent medium with the microbe-
inhibiting extracellular
product. The resulting dry composition can be applied directly onto a
conventional sanitary
product.
The manufacture of a composition comprising the absorbent and the
extracellular product
involves admixing a microbe-inhibiting amount extracellular product prepared
as described
herein, typically in the ratio of 0.1 to 1 ml of supernatant per gram of
absorbent, and thereafter
drying the absorbent medium to form the microbe-inhibiting absorbent
composition.
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The absorbent composition may further contain a degradation-enhancing bacteria
as
described herein in amounts similar to the amounts used in a sanitary product
on a weight and
volume basis. The absorbent composition may also contain a bifidogenic
oligosaccharide as
described herein.
S A preferred composition according to the present invention is an aqueous
liquid
absorbent composition that comprises an aqueous liquid~absorbing medium and an
effective
amount of a viable lactic acid-producing bacteria according to the present
invention, also
referred to as an anti-microbial absorbent composition. This composition is
useful for
application directly onto a sanitary product and has both the absorbent and
microbe inhibiting
properties described herein.
The lactic acid-producing bacteria can be any of the various bacteria
described herein,
with lactic acid bacteria preferred, and B. coagulans being particularly
preferred. The bacteria is
typically provided in the composition in the form of dried cells, a dried cell
mass or as spores in
powder, and can also be formulated into a liquid, paste, powder, granule or
pellet formulation.
1 S An absorbent anti-microbial composition typically contains about 1 OZ to
1014 CFU viable
probiotic acid bacteria per cubic meter of composition, preferably contain
about 103 to 10'° CFU,
more preferably contain 103 to 106 CFU or 106 to 109 CFU, and in preferred
embodiments
contain 108 to 109 CFU per cubic meter of absorbent composition.
The aqueous liquid absorbing medium can be any of the materials described for
an
absorbent structure herein, and need not be limited. The absorbent composition
can further
comprise a bifidogenic oligosaccharide as described herein for an absorbent
product, and
typically is present in amounts of about 10 mg to 1 gm of oligosaccharide per
cc of composition,
and preferably about 100 to 500 mg oligosaccharide per cc of composition.
E. Methods for Inhibiting Microbial Infections
The claimed invention is directed at methods for increasing dermal and
mucoidal health
and inhibiting microbial infections and microbial growth associated with use
of sanitary
products. The method comprises the use of a sanitary product comprising a
viable non-
pathogenic lactic acid bacteria, which bacterial promote dermal probiosis. In
a related
embodiment, one can administer a composition of the present invention to a pre-
existing sanitary
product, and use the product. In either case, the use of the sanitary product
containing bacteria
provides contact between the tissue in which the probiotic effect is targeted
and the sanitary
product, and thereby contacts the target tissue with an effective amount of
the active probiotic
ingredients in the composition.
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The claimed invention describes methods for inhibiting dermal or epithelial
infections
comprising the steps of contacting the surface of a sanitary product of this
invention with the
skin or mucous membrane of a mammal and maintaining the contact for a time
period sufficient
to allow initiation of probiotic activity of the lactic acid bacteria or
spores in the sanitary product,
thereby inhibiting microbial growth adjacent to or on the skin or mucous
membrane contacted by
the sanitary product.
Typically, the surface of the sanitary product used in the present method is
present on a
flexible article selected from the group consisting of a diaper, pliable
material for wiping skin or
a mucous membrane, dermal patch, adhesive tape, absorbent pad, clothing,
tampon, panty
protector, incontinence guard, sanitary napkin or the like product.
The method can be practiced to inhibit any of a variety of infections and/or
irritations
known to arise upon use of a sanitary product, including but not limited to
diaper rash, eczema,
incontinence, menstruation, fluid discharges from wounds and other
dermal/epithelial infections
or inflammations caused by opportunistic microbial pathogens that overgrow as
a result of the
irritated or inflamed skin/epithelial tissue, dermatitis, eczema, skin
allergies, and the like due to
use of the sanitary product.
Typically, the mammal is a human, although the methods and compositions of the
invention can be applied to any mammal which would require the use of a
sanitary product
where dermal infections/inflammations could be problematic.
Generally, the non-pathogenic lactic acid bacteria is used according to the
present
invention by applying a sanitary product to a tissue, which product already
contains an effective
amount of the bacteria incorporated into portions of the sanitary product.
Alternatively, a
composition of the bacterial may be applied to the sanitary product prior to
use. Administration
of a composition of this invention to a sanitary product is preferably made
using a gel,
suspension, spray, powder or semi-solid formulation containing viable
bacteria, bacterial spores
and/or probiotic extracellular product, all formulated using "good
manufacturing practice"
(GMP) methods well known in the art. Administration comprises use of typically
0.001 (i. e.,
10,000 CFU) to 10 billion colony forming units (CFU) of viable bacteria or
spore applied to a
unit of sanitary product, although lesser or greater amounts may also be used.
Application is
preferably by way of spray-drying a spore/bacteria/extracellular product
liquid suspension onto
the sanitary product's absorbent structure, preferably onto the region of the
sanitary product
which directly contacts the dermal/epithelial tissue upon use.
Upon use of the sanitary product, the absorbent structure acquires a body
fluid, such as
urine, excess fluid of fecal matter, blood, tissue exudate, pus, and the like
depending upon the
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type of sanitary product, whereupon the fluid comes into contact with the
lactic acid bacteria
present in the sanitary product. The bacteria germinates or is motivated to
grow as a result of the
contact with the body fluid, and using the electrolytes and substrates present
in the body fluid
proliferates. Upon growth, the bacterial produces metabolites that are
effective in mitigating
mycotic growth and other microbial pathogens. In addition, the growth of the
bacterial feeds on
the body fluid waste, and produces enzymes with facilitate degradation of the
sanitary product.
After the sanitary product is removed and disposed into a landfill, the
bacterial continue to grow
and degrade the waste and materials present in the construction of the
sanitary product.
In preferred embodiments where a bifidogenic oligosaccharide is included in a
composition of this invention, such as FOS, there is a synergy provided in the
form of a selective
food supply, as described herein, resulting in selective growth of acid
bacteria over food supply-
driven growth of pathogenic bacterial.
The method comprises administration of a composition of this invention
containing the
active ingredients to a human or animal in various dosage regimens as
described herein to
achieve the nutritional result. The method is typically practiced on any
animal where inhibiting
microbial infection is desired. Typically, a human in the preferred user of a
sanitary product or
composition according to the present invention, although the invention can be
practiced on any
mammal. The mammal can be any livestock or zoological specimen where such
inhibition of
parasites/pathogens provides economic and health benefits. Any animal can
benefit by the
claimed methods, including horses, cows, sheep, goats, pigs, and the like
domesticated animals.
Other purposes are readily apparent to one skilled in the arts of sanitary
products.
In carrying out the methods of the invention, it is appreciated that there are
multiple
benefits and advantages. In particular, it is noted that the presence of the
lactic acid-producing
bacteria will promote degradation of used sanitary products, when used either
alone or in
combination with other bacteria. In addition, as shown herein, viable lactic
acid bacterial growth
will exhibit a beneficial probiotic effect onto the skin or epithelial tissue
adjacent to the sanitary
product in use by out-competing dermal/epithelial pathogens, and by production
of extracellular
metabolites that inhibit dermal/epithelial pathogens.
In a related embodiment, the invention contemplates a method for enhancing
biodegradation of a sanitary product comprising the step of providing an
inoculum of viable
non-pathogenic lactic acid-producing bacteria according to this invention into
the sanitary
product, and contacting the sanitary product to the body tissue for use as
prescribed/intended for
the sanitary product. The presence of the provided inoculum in the used
sanitary product
together with the collected body fluids/exudates provides an environment for
bacterial growth,
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which facilitates the breakdown of the various components of the sanitary
product, including the
waste material containing in the disposed sanitary product and the various
components making
up the sanitary product, such as cellulose, petroleum hydrocarbon polymers,
natural and
synthetic fibers and adhesives, and the like materials described herein for
constructing a sanitary
product.
As noted above, enhancement of degradation of a sanitary product upon disposal
can be
facilitated by use of a lactic acid bacteria. In a particular embodiment, the
method comprises the
use of a non-pathogenic lactic acid bacteria and comprises the use of one or
more additional
species of bacteria designed to enhance degradation. These additional bacteria
are non-
pathogenic insofar as they are included in the sanitary product, but are not
necessarily probiotic
in terms of inhibiting dermal or epithelial microbial infections. Rather,
these additional bacteria
are included solely for the ability to enhance degradation, and are referred
collectively as
"degradation-enhancing" bacteria.
F. Articles of Manufacture
The invention also contemplates various articles of manufacture which utilize
the
beneficial aspects of the present invention by combination of a composition
with various medical
or personal hygiene devices so as to reduce or prevent microbial infections
associated with the
use of these devices. The invention comprises compositions of a probiotic
lactic acid bacteria,
preferably a lactic acid bacteria, and more preferably a Bacillus species
and/or isolated
B. coagulans active agent, applied to a solid surface or impregnated into a
solid matrix of any
device or article of manufacture that is intended to be in contact with skin
or a mucous
membrane. Preferably the solid surface is a flexible article than can be worn
on or wiped on the
skin or mucous membrane. More preferably, when the flexible item carrying the
acid bacteria is
to be worn on the skin it includes a means for attaching the article to the
skin such as, for
example, an adhesive layer, interengaging hook and pile (i.e., Velcro~)
connectors, or other well
known means of attachment such as ties, snap closures, elastic, buttons, and
the like.
Many different types of absorbent products having absorbent structures are
well known in
the art, and can include diapers, towelettes (e.g., baby wipes or feminine
hygiene towelettes),
sanitary napkins, tampons, panty protectors, dermal patches, adhesive tape,
bandages, wound or
sore dressings, absorbent pads, incontinence guards, bed sheets or protectors,
saliva absorbent,
articles of clothing (e.g., underclothes, sleeping apparel), bath towels, wash
cloths, and the like.
Thus, the claimed invention describes an absorbent product comprising an
aqueous liquid
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absorbent structure and an effective amount of viable non-pathogenic acid
bacteria according to
said invention.
Absorbent structures in sanitary products (absorbent products) are typically
produced by
fluffing cellulosic or other fibrous pulp into a roll, bale or sheet for
instance, to form a pulp mat,
sometimes admixed with so-called superabsorbent materials in the pulp mat. The
superabsorbent
materials are typically polymeric formulations capable of absorbing many times
their own
weight'of water of body fluid, and are well known in the art. The pulp mat is
typically
compressed so as to enhance its fluid-wicking ability and also in order to
reduce pulp body bulk,
and therewith obtain an article which is as compact as possible to achieve the
absorbent
properties desired in the particular sanitary product.
The absorbent structure may also include other constituents, for example,
components
which will improve fluid acquisition properties, fluid-wicking properties,
fluid retention
properties, and the like well known in the art. Other included constituents
include components
which increase coherent strength (i.e., the ability to withstand deformation
during use). The
absorbent structure may contain fibrous woven, knitted or non woven materials,
occlusive or
non-occlusive films or membranes, granules, pellets or aggregates of absorbent
material,
synthetic polymer fibers, films, membranes and foams (e.g., nylon,
polytetrafluoroethylene
(PTFE, such as Teflon or Gor-TeX ), polystyrene, polycarbonate,
polyvinylchloride and
polysulphone). All of these forms are well known in the art and include, for
example, knitted or
woven fabrics, non-woven fabrics such as felt and batting, fiber balls of
cotton, rayon, cellulose
or synthetic fibers, and the like materials.
The fibers can be natural fibers, including but not limited to: wool, silk,
cotton, cellulosic
fiber, and the like natural fibers. Natural polymers based on polysaccharide
can also be used,
including, but not limited to: modified cellulose and cellulose derivatives
(e.g., alkyl-,
hydroxyalkyl-, carboxymethylcellulose); gum resins (e.g., guar gum, locust
bean gum, tragacanth
gum, gum arabic, pectin, etc.); starch and starch derivatives (e.g., corn
starch, grain starch, potato
starch, amylose, amylopectin, dextrin, dextran, modified starch, hydroxy-ethyl
starch, cationic
starch, starch graft polymers, and the like polymers). The fibers can be
synthetic fibers,
including, but not limited to: polyester, polyolefin, polyamide, polyvinyl
alcohol, polyvinyl
acetate, polyvinyl chloride, polyvinyl urea, polyurethane, polyurea,
polyacrylonitrile, as well as
copolymers of these polymers, and the like synthetic fibers.
The absorbent product can be formatted into a mufti layer configuration,
having an
absorbent structure layer, a fluid permeable top layer which allows wicking of
fluid but is itself
non-wettable due to its structural composition (e.g., synthetic fiber
construction), and a fluid-
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impermeable bottom layer (i.e., back sheet) which prevents absorbed fluid to
pass from the
absorbent structure layer to the adjacent tissues of the user when contacted
by the absorbent
product during use. Such layered configurations are well known in the diaper
and panty liner
arts, and need not be described in detail.
The absorbent product typically contains about 102 to 1014 CFU of viable
probiotic acid
bacteria per cubic meter (M3) of absorbent product, and is typically dispersed
in the form of
cells, dried cell mass or spores, with spores being the particularly preferred
format. Preferably,
the product will contain about 103 to 101° CFU per M3 of absorbent
product, and may have 103 to
106 CFU per M3 or 106 to 109 CFU per M3, and preferably will have 108 to 109
CFU per M2 of
absorbent product, although these amounts can vary depending upon the specific
application,
product formulation and intended use.
The absorbent product may further contain a bifidogenic oligosaccharide
dispersed
therein as described for a composition according to the present invention,
typically in amounts of
from about 10 milligrams (mg) to 1 gram per cubic centimeter (cc) of absorbent
structure, and
preferably about 100 to S00 mg of oligosaccharide per cc of structure.
The absorbent product may further contain a degradation-enhancing bacteria
according to
the present invention, as described herein. Where the degradation-enhancing
bacteria is different
from the probiotic lactic acid-producing bacteria, the degradation enhancing
bacteria can be
incorporated into the absorbent product is regions of the product designed to
keep the bacteria
away from the skin or tissues of the mammal during use, and designed for
release or access to
the waste products. Typically, an absorbent product will contain from 104 to
1014 CFU of viable
degradation-enhancing bacteria per unit of sanitary product, preferably about
from 10' to lOlo
CFU per unit, and more preferably about from 108 to 10~ CFU per unit.
A composition containing a lactic acid bacteria of this invention can be
applied to any of
a variety of regions of an absorbent product of the present invention
including the moisture
barner (i.e., the "stay-dry lining"), the absorbent structure (e.g., moisture
absorbing polymer),
coated onto the external surface that contacts the skin or epithelial tissue,
in capsules which are
sealed-off until wetted for slow release of bacteria/spores, or combinations
thereof. The bacteria
can be presented as a spore, a dry or lyophilized cell mass, a stabilized gel
or paste, a dry
powder, or as a component of the gel polymer that comprises the moisture
barrier system.
In addition, insofar as the Bacillus coagulans extracellular product (i.e.,
the "isolated
agent") can be used to inhibit microbial pathogens, the invention contemplates
the use of the
extracellular product in place of or in combination with a viable acid
bacteria in any of the
sanitary products described herein.
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The lactic acid bacteria and/or a B. coagulans-isolated active agent can be
applied to the
solid surface using any of a variety of known methods including, for example,
applying a
powder, spray drying the probiotic onto the material or soaking the material
in a solution
containing the probiotic and then using the wetted material or drying the
material before use.
Porous material may contain the Bacillus and/or the isolated active agent in
the pores or
interstices of the solid material. The Bacillus andlor the isolated active
agent can be attached by
adhesion, such as by attachment to an adhesive layer that is then applied to
the skin (e.g., in a
bandage or dermal patch). The Bacillus and/or the isolated active agent can be
impregnated into
the solid material during the manufacturing process of the flexible article
(e.g., added to a
synthetic composition before or during the polymerization process). The
pressure and heat
resistance of Bacillus spores makes them particularly suitable for
incorporation into the material
during manufacturing.
Any of the solid materials carrying Bacillus and/or the isolated active agent
can be
packaged individually or in groups, suitable for holding the treated material
using standard
packaging materials (e.g., in a shrink wrapper, sealed packet, protective
wrapper or dispensing
container suitable for holding dry or wet materials) The article of
manufacture can have applied
thereon any of the additional/optional components of a composition of this
invention, including
earners, disinfectants, antibacterial agents, salts, FOS, and the like. In
particular, the absorbent
product can include as a component part of the absorbent structure inert
ingredients, neutral
filling agents ,and the like. Exemplary neutral filling agents include peat,
sand, clay, garden
mold, ground shells of nuts or pomaceous fruit, wood flour, chitin-containing
flour, and the like
well known materials.
Any of a variety of methods for placing the composition onto a subject article
can be
used, and therefor the invention need not be so limited. However, preferred
methods include a
"spray-dry" method in which the material is exposed in a low humidity chamber
to an atomized
mix containing a liquid composition, where the chamber is subsequently exposed
to about
80-110°F to dry the liquid, thereby impregnating the material of the
article with the components
of the composition. A typical load is from 105 to 109 CFU of bacteria/spores
per ml of atomizing
mix, to place that same amount on about one square inch of external surface of
fibrous
carner/article material. The dry article is then ready for storage in a
sterile package for use.
In one embodiment, the invention contemplates the use of viable probiotic acid
bacteria
in absorbent structures which structures are incorporated into an absorbent
product as described
herein. Thus, the invention describes the use of an effective amount of
probiotic lactic acid
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WO 00/61201 PCT/US00/10222
bacteria where the bacteria is dispersed or incorporated in the form of dried
cells or cell mass or
as spores into the absorbent structure of an absorbent product according to
the invention.
G. Therapeutic Systems for Inhibiting Infections Associate With
Use of Sanitary Products
The claimed invention further contemplates a system for inhibiting dermal or
epithelial
infection associated with use of a sanitary product comprising a container
comprising label and
an absorbent composition according to the present invention, wherein said
label comprises
instructions for application of the absorbent composition for inhibiting
microbial infections
during use of the sanitary product.
Typically, the system is present in the form of a package containing a
composition or
absorbent product of this invention, or in combination with packaging
material. The packaging
material includes a label or instructions for use of the components of the
package. The
instructions indicate the contemplated use of the package component as
described herein for the
methods or compositions of the invention.
For example, a system can comprise one or more unit dosages of a composition
according to the invention. Alternatively, the system can contain bulk
quantities of a
composition. The label contains instructions for using the composition or
absorbent product as
appropriate, and may include information regarding storage of the composition
or absorbent
product, application methods, health indications, methods for disposal, and
the like information.
Unless defined otherwise, all scientific and technical terms used herein have
the same
meaning as commonly understood by those skilled in the relevant art. Unless
mentioned
otherwise, the techniques employed or contemplated herein are standard
methodologies well
known to one of ordinary skill in the art. The Examples of embodiments are for
illustration only.
EXAMPLES
The following Examples relating to this invention are illustrative and should
not, of
course, be construed as specifically limiting the invention. Moreover, such
variations of the
invention, now known or later developed, which would be within the purview of
one skilled in
the art are to be considered to fall within the scope of the present invention
hereinafter claimed.
Example 1: Preparation of Bacillus coagulans Cultures
Bacillus coagulans Hammer bacteria (ATCC Accession No. 31284) was inoculated
and
grown to a cell density of about 108 to 109 cells/ml in nutrient broth
containing 5 g Peptone, 3 g
Meat extract,10-30 mg MnS04, and 1,000 ml distilled water, adjusted to pH 7.0,
using a standard
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airlift fermentation vessel at 30°C. The range of MnS04 acceptable for
sporulation is 1 mg/1 to
1 g/1. The vegetative cells can actively reproduce up to 45°C, and the
spores are stable up to
90°C. After fermentation, the B. coagulans bacterial cells or spores
are collected using standard
methods (e.g., filtration, centrifugation) and the collected cells and spores
can be lyophilized,
S spray dried, air dried or frozen: As described herein, the supernatant from
the cell culture can be
collected and used as an extracellular agent secreted by B. coagulans which
has antimicrobial
activity useful in a formulation of this invention.
A typical yield from the above culture is in the range of about 109 to
101° viable spores
and more typically about 100 to 150 billion cells/spores per gram before
drying. Spores maintain
at least 90% viability after drying when stored at room temperature for up to
ten years, and thus
the effective shelf life of a composition containing B. coagulans Hammer
spores at room
temperature is about 10 years.
Example 2: Preparation of Bacillus coagulans Spores
A culture of dried B. coagulans spores was alternately prepared as follows.
Ten million
1 S spores were inoculated into a one liter culture containing 24 g potato
dextrose broth, 10 g of
enzymic-digest of poultry and fish tissue, 5 g of FOS and 10 g MnS04. The
culture was
maintained for 72 hours under a high oxygen environment at 37°C to
produce culture having
about 150 billion cells per gram of culture. Thereafter, the culture was
filtered to remove culture
medium liquid, and the bacterial pellet was resuspended in water and freeze-
dried. The freeze-
dried powder is then ground to a fine powder using standard good manufacturing
practice
(GMP).
Example 3: Preparation of Bacillus coagulans Extracellular Products
A one liter culture of B. coagulans was prepared as described in Example 1.
The culture
was maintained for 5 days as described, at which time FOS was added at 5
g/liter, and the culture
was continued. 20 ml of carrot pulp was then added at day 7, and the culture
was harvested when
the culture became saturated (i.e., no substantial cell division). The culture
was first autoclaved
for 30 minutes at 250°F, and then centrifuged at 4000 rpm for 15 mm.
The resulting supernatant
was collected and filtered in a Buchner funnel through a 0.8 micron (gym)
filter, and the filtrate
(i.e., the "pass-through") was collected and further filtered through a 0.2 ~m
Nalge vacuum
filter. The resulting pass-through was collected (about 900 ml) to form a
liquid containing an
extracellular product, and used in inhibition studies.
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WO 00/61201 PCT/US00/10222
Following the assay described in Example 5, except using Candida albicans, 1
ml of the
above-produced extracellular product was added to the test plate in place of
live B. coagulans.
After the same culturing time, a zone of inhibition of about 10 to 25
millimeters was observed,
indicating a potent antimicrobial activity of "excellent" quality, using the
terminology set forth in
S Example 5.
Example 4: Formulations
Formulation 1: (Powder for Application to Sanitary Product)
B. coagulans 250,000,000 spores (approximately 17.5 mg)
Fructo-oligosaccharides (FOS) 100 mg
Micro-crystalline cellulose (MCC) 372.5 mg
Formulation 2: (Diaper-Sanitary Product)
Lined diaper containing a composition of
B. coagulans 1 billion spores (approximately 70 mg)
Fructo-oligosaccharides (FOS) 500 mg
Micro-crystalline cellulose (MCC) 1 g
dispersed uniformly in the absorbent fibers of the diaper.
Formulation 3: (Diaper - Sanitary Product)
Lined diaper containing a composition of:
B. coagulans 1 billion spores (approximately 70 mg)
B. licheniformis 250 million spores
B. subtilis 250 million spores
B. pumilis 250 million spores
B. megaterium 250 million spores
Fructo-oligosaccharides (FOS) 500 mg
Micro-crystalline cellulose (MCC) 1 g
dispersed uniformly in the absorbent fibers of the diaper.
Example 5: Antimicrobial Activity of Bacillus coagulaus
The ability of B. coagulans to inhibit bacterial pathogens was demonstrated
using an
in vitro assay. The assay is part of a standard bacterial pathogen screen
(U.S. Food and Drug
Administration) and is commercially available on solid support disks (DIFCO~
BACTROL~ disk
set). In the assay, potato-dextrose plates (DIFCO~) were prepared using
standard procedures and
were inoculated individually with a confluent bed 1.5 x 106 of each species of
bacteria tested.
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Inhibition by B. coagulans was tested by placing on the plate about 1.5 x 106
CFU in 10 pl of
broth or buffer, plated directly in the center of the potato-dextrose plate
with one test locus of
about 8 mm in diameter per plate. A minimum of three test loci were used for
each assay. The
negative control was comprised of a-10 ~l drop of a sterile saline solution,
whereas the positive
S control was comprised of a 10 p,1 volume of glutaraldehyde. The plates were
then incubated for
about 18 hr at 30°C when the zone of inhibition was measured.
As used herein, "excellent inhibition" means the zone was 10 mm or greater in
diameter;
and "good inhibition" means the zone was greater than 2 mm in diameter, but
less than 10 mm in
diameter.
No inhibition was seen with the negative control and excellent inhibition
(about 16.2 mm
diameter, average of three tests) was seen with the positive control. For the
enteric organisms
tested, Clostridium species and E. coli, excellent inhibition by B. coagulans
was seen. For the
Clostridium species, C. perfringens, C. difficile, C. botulinum, C.
tributrycum and C. sporogenes,
the zone of inhibition was consistently greater than 15 mm in diameter.
Similarly, excellent
inhibition was also seen for the opportunistic pathogens Pseudomonas
aeruginosa,
Staphylococcus aureus, and Candida species.
The present invention has been described in the above Examples using a variety
of
formulations, although it should be apparent that various other carrier agents
that are compatible
with the probiotic compositions may be substituted in the examples to give
similar results.
Accordingly, the present invention may be embodied in other specific forms
without departing
from it in spirit. The Examples are to be considered in all respects only as
illustrative and not as
restrictive, and the scope of the invention is indicated by the claims that
follow. All
modifications which come within the meaning and range of the lawful
equivalency of the claims
are to be embraced within their scope.
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