Classifications

• • C—CHEMISTRY; METALLURGY
  • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F1/00—Treatment of water, waste water, or sewage
  • C02F1/50—Treatment of water, waste water, or sewage by addition or application of a germicide or by oligodynamic treatment
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/04—Antibacterial agents
• • C—CHEMISTRY; METALLURGY
  • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F1/00—Treatment of water, waste water, or sewage
  • C02F1/28—Treatment of water, waste water, or sewage by sorption
  • C02F1/286—Treatment of water, waste water, or sewage by sorption using natural organic sorbents or derivatives thereof
• • C—CHEMISTRY; METALLURGY
  • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
  • C02F2103/42—Nature of the water, waste water, sewage or sludge to be treated from bathing facilities, e.g. swimming pools

Definitions

• This invention relates to methods of inhibiting microorganism growth and to methods of water treatment using moss, particularly sphagnum moss.
• Various biological control agents are known in the art for the control of microorganism growth. For example, it is desirable to inhibit fungal growth in food products, such as when grain is stored, and to inhibit bacterial growth in packaged foods, such as raw chicken, hi other fluid absorbing products such as bandages or feminine hygiene products, it is desirable to include antimicrobial compositions so that pathological organisms cannot flourish.
• Water treatment is another area in which chemical, biological, and radiation treatments have been used to control or prevent bacterial growth.
• There are many types of water treatment systems such as filtration and cleaning systems for swimming pools and aquariums. Many of these systems filter the water to remove suspended matter and reduce the cloudy appearance of the water. Preventing bacterial growth in water and removing contaminants from water are significant industrial, as well as household, problems.
• industrial effluent should be cleaned to remove toxic compounds as well as to remove bacteria before it is dumped into lakes and rivers.
• Containers of water such as swimming pools, hot tubs, aquariums and the like must be kept clean to prevent the water from becoming cloudy and/or the container walls from becoming slimy.
• the water may be treated by active means such as a filter to remove particles and bacteria, and it may also be treated by passive means whereby a biocide is placed in a container and floated in the water. It is common to use chemical means to keep the water clean and reduce bacterial growth.
• Ultraviolet light, chlorination, bromination, treatment with ions of copper and silver as well as treatment with ozone can be used to treat and/or disinfect water.
• Sphagnum moss is a generic expression that designates a range of botanical species that co-exist in a sphagnous bog. It should be noted that “peat moss” refers generally to a decomposed or composted sphagnum moss.
• Sphagnum moss is commonly harvested for use in various products.
• the petals, and not the stems, of the moss preferably may be harvested.
• large pieces of plant material roots, twigs, etc.
• the moss may be processed further after harvesting by forming an aqueous slurry to extract very fine particles. Water is removed from the slurry and the moss is dried. The moss may be compressed prior to packaging or shipment.
• Various additives may be used to alter the absorption characteristics or mechanical properties of the moss. Because sphagnum moss is readily available and relatively inexpensive, it has been used in a variety of products, primarily for the absorption of fluids.
• the invention provides a method of inhibiting microorganism growth comprising contacting a substance susceptible to microorganism growth with an amount of a non-decomposed moss effective to inhibit microorganism growth, wherein the moss is selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof.
• the invention provides a method of inhibiting microorganism growth comprising placing an amount of a non-decomposed moss effective to inhibit microorganism growth in a carrier and contacting the carrier with a substance susceptible to microorganism growth, wherein the moss is selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof.
• the invention provides a method of preparing moss for use in inhibiting microorganism growth comprising: (i) drying non-decomposed moss; and (ii) sterilizing the moss.
• the invention provides a method of preparing moss for use in inhibiting bacterial growth comprising: (i) contacting non-decomposed moss with an acidic solution; and (ii) drying the moss.
• the invention provides a kit comprising sterilized, non-decomposed moss and a carrier.
• the invention provides a method of inhibiting microorganism growth comprising contacting water susceptible to microorganism growth with an amount of a non-decomposed moss effective to inhibit microorganism growth, wherein the moss is selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof, and periodically shocking the water with an appropriate chemical agent.
• the invention provides a method of treating water comprising contacting water with an amount of a non-decomposed moss effective to remove cations other than hydrogen ions from the water, wherein the moss is selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof.
• the invention also provides a method of treating water comprising placing in a carrier an amount of a non-decomposed moss effective to remove cations other than hydrogen ions from the water and contacting the carrier with water, wherein the moss is selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof.
• the invention provides a device for use in water comprising: (i) a carrier having an interior cavity and one or more openings allowing ingress to and egress from the interior cavity; and (ii) a moss contained within the interior cavity, wherein the carrier completely encloses the moss.
• the invention provides a method of inhibiting microorganism growth comprising placing in water susceptible to bacterial growth a device for inhibiting microorganism growth in water, the device comprising: (i) a carrier having an interior cavity and one or more openings allowing ingress to and egress from the interior cavity; and (ii) a moss contained within the interior cavity, wherein the carrier completely encloses the moss, and wherein the device comprises an amount of the moss effective to inhibit microorganism growth in the water.
• the invention provides a kit comprising sterilized, non-decomposed moss and a device for use in water comprising a carrier having an interior cavity and one or more openings allowing ingress to and egress from the interior cavity, wherein the interior cavity can completely enclose the moss.
• the invention provides a method of inhibiting microorganism growth comprising placing in water susceptible to microorganism growth a device for inhibiting microorganism growth in water, the device comprising: (i) a carrier having an interior cavity and one or more openings allowing ingress to and egress from the interior cavity; and (ii) a moss contained within the interior cavity, wherein the carrier completely encloses the moss, wherein the device comprises an amount of the moss effective to inhibit microorganism growth in the water, and periodically shocking the water with an appropriate chemical agent.
• the invention provides a method of treating water comprising placing in water a device comprising: (i) a carrier having an interior cavity and one or more openings allowing ingress to and egress from the interior cavity; and (ii) a moss contained within the interior cavity, wherein the carrier completely encloses the moss, and wherein the device comprises an amount of the moss effective to remove cations other than hydrogen ions from the water.
• Figure 1A illustrates a perspective view of one embodiment of a device that can be used in the invention.
• Figure IB illustrates a side view
• FIG. 1C illustrates a cross-sectional view along line C — C of Figure IB.
• Figure 2A illustrates a perspective view of another embodiment of a device that can be used in the invention and Figure 2B shows a side view of the moss used within the device shown in Figure 2A.
• Figure 3 illustrates a perspective view of another embodiment of a device that can be used in the invention.
• sphagnum moss that can be used to inhibit microorganism growth.
• the moss can be used to confrol microorganism growth in water such as in a swimming pool, spa, aquarium, and the like. Spas are also known as whirlpools or hot tubs. It is believed that particular species of moss are particularly effective at inhibiting and/or preventing the growth of bacteria and other microorganisms.
• bacteriostatic refers to a material that inhibits the growth of bacteria
• antibacterial generally refers to a bacterial growth inhibitor. Both terms should be distinguished from "bactericidal” which refers to materials that kill bacteria upon contact.
• water treatment refers to a process by which water is kept clean, clear, and pleasant smelling in swimming pools, aquariums, whirl pool baths, hot tubs, and the like. When the water is agitated, less foaming is observed.
• the moss is believed to inhibit growth of bacterial and other microorganisms and it also may absorb compounds and substances that decrease water clarity.
• sphagnum papillosum S. papillosum
• S. cristatum sphagnum cristatum
• the moss is enclosed or encapsulated in a mesh material that prevents the moss from disintegrating in an aqueous environment.
• moss can be held in a desired place in a pool, hot tub, whirlpool bath, and the like.
• Preferred mesh materials include those comprising polymers such as nylon or polypropylene, with mesh sizes ranging from about 0.1 to 1 mm. Polymers are generally preferred because they are inexpensive and may be resistant to degradation. Suitable for use in this invention are S. papillosum, which can be harvested from bogs in northern Minnesota, U.S.A., and S. cristatum, which is commercially available as a compressed bale from Sutton's Moss of Dobson, Westland, New Zealand. These species of moss can be used by themselves or together in the devices and systems of this invention.
• the moss is cleaned to remove small particles, such as dirt, and larger debris, such as roots and leaves.
• Commercially available moss may be fumigated before it is packaged by a manufacturer in order to destroy seeds.
• the moss is cut by mechanical means into a desired size and shape.
• the moss preferably is then sterilized by autoclaving, exposure to ethylene oxide, or by other means known to one of skill in the art. Sterilization destroys living organisms in the moss and thus avoids any problems of undesirable or foreign bacteria being introduced into the environment where a device of this invention is used.
• the moss is then ready for use in a water treatment system or other applications.
• the water freatment devices described herein are preferably used in conjunction with materials that kill bacteria. This is because these environments may have large bacteria loads introduced at various times. Accordingly, standard practice is to filter the water, flush water lines, and test the water as necessary. The pH can be adjusted by using commercially available solutions.
• the water treatment devices of this invention are most desirably used in conjunction with an oxidizer, such as potassium monopersulfate, referred to as "chlorine free shock".
• Potassium monopersulfate is known to increase the efficiency of chlorine purification products, but we have found that it is also particularly effective when used with the sphagnum moss devices described above.
• the sphagnum moss of this invention can be used in any composition, material, device, or method where the inhibition of microorganisms is desirable. Uses include the inhibition of microorganism growth, the reduction and/or prevention of odors, water freatment, and confrol of mold and fungal growth; and control of fermentation.
• Such devices and materials include absorbent products, such as diaper liners, feminine hygiene products, bandages, and wound dressings. In such products, the moss can be enclosed between membranes of differing liquid transmission characteristics.
• the moss can be incorporated into polymers and used as face masks.
• the moss can be encapsulated in membranes and used in food preservation products such as packaging wraps and liners to absorb liquid and odors.
• the moss can be used in water treatment products to keep water clean in storage tanks, aquariums, swimming pools, whirlpool baths, spas, and the like, as well as in water filfration devices.
• the moss can be used for waste water and sewage freatment.
• the moss can be shaped into, for example, discs or pellets, and used to absorb water from grain and other food products.
• the moss also can be used for fermentation control (such as in liquids or grains).
• the moss can be used for the control of fungal or microorganism diseases in lawns and gardens.
• the moss can be used for mold control products such as in storage containers or ductwork linings.
• the invention provides a method of inhibiting microorganism growth comprising contacting a substance susceptible to microorganism growth with an amount of a non-decomposed moss effective to inhibit microorganism growth, wherein the moss is selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof.
• the moss can be compressed and can be in the form of strips.
• the moss can be sterilized by autoclaving, sterilized by chemical treatment, or sterilized by freatment with ethylene oxide.
• the moss can be washed with an acidic solution, especially a solution of acetic acid.
• the moss can be washed with an acidic solution and then washed with a salt solution.
• the substance susceptible to microorganism growth can be water.
• the water can be in a spa, pool, or aquarium.
• the substance susceptible to microorganism growth can be a portion of the human body such as skin, a surface wound, an internal body cavity, or the site of an internal injury.
• the substance susceptible to microorganism growth can be grain.
• the invention provides a method of inhibiting microorganism growth comprising placing an amount of a non-decomposed moss effective to inhibit microorganism growth in a carrier and contacting the carrier with a substance susceptible to microorganism growth, wherein the moss is selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof.
• the carrier can be a device that is implanted into the human body, a polymer matrix, a biomafrix, or one or more membranes.
• the invention provides a method of preparing moss for use in inhibiting microorganism growth comprising: (i) drying non-decomposed moss; and (ii) sterilizing the moss.
• the method can further comprising compressing the moss, compressing the moss and cutting the moss into strips, sterilizing the moss by autoclaving, chemical treatment, or treatment with ethylene oxide.
• the moss can be sphagnum moss.
• the moss can be selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof.
• the invention provides a method of preparing moss for use in inhibiting bacterial growth comprising: (i) contacting non-decomposed moss with an acidic solution; and (ii) drying the moss.
• the method can comprise contacting the non- decomposed moss with a salt solution after step (i).
• the acidic solution is a solution of acetic acid.
• the moss can be sphagnum moss.
• the moss can be selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof.
• the invention provides a kit comprising sterilized, non-decomposed moss and a carrier.
• the moss can be compressed and can be in the form of strips.
• the invention provides a method of inhibiting microorganism growth comprising contacting water susceptible to microorganism growth with an amount of a non-decomposed moss effective to inhibit microorganism growth, wherein the moss is selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof, and periodically shocking the water with an appropriate chemical agent.
• the chemical agent can be potassium monopersulfate.
• the invention provides a method of treating water comprising contacting water with an amount of a non-decomposed moss effective to remove cations other than hydrogen ions from the water, wherein the moss is selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof.
• the cations can be calcium or iron ions, and substantially all of the calcium or iron ions can be removed from the water.
• the moss can be compressed and can be in the form of strips.
• the moss can be sterilized by autoclaving, sterilized by chemical treatment, or sterilized by freatment with ethylene oxide.
• the moss can be washed with an acidic solution, especially a solution of acetic acid.
• the moss can be washed with an acidic solution and then washed with a salt solution.
• the water can be in a spa, pool, or aquarium.
• the invention provides a method of treating water comprising placing in a carrier an amount of a non-decomposed moss effective to remove cations other than hydrogen ions from the water and contacting the carrier with water, wherein the moss is selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof.
• the invention provides a device for use in water comprising: (i) a carrier having an interior cavity and one or more openings allowing ingress to and egress from the interior cavity; and (ii) a moss contained within the interior cavity, wherein the carrier completely encloses the moss.
• the carrier can comprise a float.
• the carrier can comprise a float and a cylindrical portion beneath the float, the cylindrical portion having an interior cavity and one or more openings allowing ingress to and egress from the interior cavity.
• the moss can be enclosed within a mesh bag.
• the carrier can comprise one or more weights.
• the moss can be non-decomposed moss.
• the moss can be sphagnum moss.
• the moss can be selected from the group consisting of sphagnum papillosum, sphagnum cristatum, and mixtures thereof.
• the moss can be compressed and can be in the form of strips.
• the moss can be sterilized by autoclaving, sterilized by chemical treatment, or sterilized by treatment with ethylene oxide.
• the moss can be washed with an acidic solution, especially a solution of acetic acid.
• the moss can be washed with an acidic solution and then washed with a salt solution.
• the invention provides a method of inhibiting microorganism growth comprising placing in water susceptible to microorganism growth a device for inhibiting microorganism growth in water, the device comprising: (i) a carrier having an interior cavity and one or more openings allowing ingress to and egress from the interior cavity; and (ii) a moss contained within the interior cavity, wherein the carrier completely encloses the moss, and wherein the device comprises an amount of the moss effective to inhibit microorganism growth in the water.
• the water can be in a spa, pool, or aquarium.
• the water can be pumped through the device.
• the invention provides a kit comprising sterilized, non-decomposed moss and a device for use in water comprising a carrier having an interior cavity and one or more openings allowing ingress to and egress from the interior cavity, wherein the interior cavity can completely enclose the moss.
• the kit can comprise one or more pH test strips and/or potassium monopersulfate.
• the invention provides a method of inhibiting microorganism growth comprising placing in water susceptible to microorganism growth a device for inhibiting microorganism growth in water, the device comprising: (i) a carrier having an interior cavity and one or more openings allowing ingress to and egress from the interior cavity; and (ii) a moss contained within the interior cavity, wherein the carrier completely encloses the moss, wherein the device comprises an amount of the moss effective to inhibit microorganism growth in the water, and periodically shocking the water with an appropriate chemical agent.
• the chemical agent can be potassium monopersulfate.
• the invention provides a method of treating water comprising placing in water a device comprising: (i) a carrier having an interior cavity and one or more openings allowing ingress to and egress from the interior cavity; and (ii) a moss contained within the interior cavity, wherein the carrier completely encloses the moss, and wherein the device comprises an amount of the moss effective to remove cations other than hydrogen ions from the water.
• the cations can be calcium or iron ions, and substantially all of the calcium or iron ions can be removed from the water.
• the moss can be compressed and can be in the form of strips.
• the moss can be sterilized by autoclaving, sterilized by chemical freatment, or sterilized by treatment with ethylene oxide.
• the moss can be washed with an acidic solution, especially a solution of acetic acid.
• the moss can be washed with an acidic solution and then washed with a salt solution.
• the water can be in a spa, pool, or aquarium.
• Figures 1A to 1C illustrate a suitable device of this invention.
• Figure 1A shows device 10 floating in water and
• Figures IB and 1C shows side and cross sectional views, respectively.
• Device 10 is adapted to receive a segment of compressed sphagnum moss 15 that has been cut into a desired dimension.
• the moss is shown in phantom in Figure 1A and IB.
• a convenient dimension for the - moss used in device 10 is about 6 x % x Vi inches (15.2 x 0.63 x 0.63 cm). A piece of moss this size weighs about 5 grams. Moss 15 is enclosed in nylon mesh 16, sized to permit the compressed moss to expand. The mesh size is such that it will retain even small particles of moss and prevent it from breaking apart and floating away.
• Device 10 comprises a plastic material that is impact resistant, does not dissolve in water, and can be shaped into a desired shape.
• Device 10 is commercially available as a "floater" from MP Industries of Huntington Beach, CA. It should be noted that floaters of this type are commonly used with pellets or discs of pool cleaning agents, such as those containing chlorine.
• Device 10 has been adapted for use with sphagnum moss by adding holes to facilitate passage of water into the device.
• Device 10 comprises float portion 20 and flow through portion 30.
• Float portion 20 is cylindrical, and may be any desired dimension, though typically it is larger in diameter than flow-through portion 30.
• a useful dimension for the float portion is about 5 inches (12.7 cm) in diameter.
• Flow-through portion 30 is a two-part elongated cylinder having core or hollow center 32.
• First part 33 is attached to floatation portion 20 and is provided with screw threads onto which second part 35 affixes. In this way the length of the flow-through portion can be changed.
• Second part 35 is fixed in position by means of adjustable collar 34.
• Second part 35 also has removable cap 37, which is weighted so that device 10 floats in the water as illustrated in Figure 1.
• Slots 38 and holes 39 permit water to flow through the cylinder.
• the slots and holes may be any desired dimension and can be positioned as desired.
• a useful length of the flow through portion is about 7 inches (17.8 cm).
• Cap 37 is removable so that the desired size of the sphagnum moss can be inserted into portion 30. Once exposed to water, the compressed moss expands. The density of expanded moss is such that water can flow through it.
• Device 10 is sufficient to treat up to about 350 gallons of water (for example, in a whirlpool or spa) for up to 30 days.
• Figure 2A illustrates device 50 floating in water.
• Device 50 comprises cylindrical portion 60 having core or hollow center 62. Slots 64 and holes 66 permit water to enter the hollow center.
• Moss 55 shown in phantom in Figure 2A, is encapsulated by mesh 52, as most clearly shown in Figure 2B. The moss expands when in contact with the water, filling hollow center 62. Cylindrical portion 60 is shown sealed at one end, with removable cap 57 at the other end. Cap 57 may be weighted so that the maximum length of device 60 stays in contact with the water.
• Figure 3 shows device 70 attached to wall W of a swimming pool, aquarium, hot tub, or the like.
• Moss 75 is encapsulated by mesh 72 and the mesh is affixed to bracket 77.
• the mesh is of a sufficient size that particles or fragments of moss will stay within the mesh.
• the bracket hangs from the wall and the device can remain fixed at this location.
• device 70 could lie on the bottom of the pool or tub. It could be affixed there or could be held down by a weight. It also could be placed in-line with a filter.
• Example 1 S. papillosum moss, harvested from northern Minnesota, and was prepared for bacterial inhibition testing. The moss species was validated by the University of Minnesota and again upon receipt. All samples were placed in plastic bags. All raw moss was stored at 4°C until processed by lab personnel. All pre-dried outside moss samples were stored at room temperature until processed.
• Blender 1.25 L capacity (commercially available as Osterizer® from Oster)
• Distilled Water available from Premium Water, Inc.)
• Tissue Sieve 1 cup capacity (commercially available as Cellector® E-C Apparatus Corp.)
• 1 L Glass Beaker commercially available as Pyrex®
• Sterile Polystyrene Pefri Dishes 100 x 15 mm commercially available from Falcon
• f) Sterile Polystyrene Pefri Dishes 150 x 15 mm commercially available from VWR
• Autoclave commercially available from Market Forge
• Metal Lab Scoop (16.5cm) / commercially available as Adison Tissue
• the moss was brought to the laminar flow hood and carefully placed in labeled, pre-weighed Pefri dishes using a sterile lab scoop and forceps. Special care was taken not contaminate the moss and to pack each dish in a uniform manner. Once packed, the dishes remain uncovered for at least 72 hours until the moss was dry. The dried dishes were covered and kept in the flow hood until used.
• Example 2 S. cristatum moss, obtained from Sutton's Moss, Canada, (harvested in New Zealand) was prepared for bacterial inhibition testing. The moss species was validated upon receipt. Handling of the moss samples was identical to that described above in Example 1.
• Example 3 This experiment determined the amount of bacterial growth in Tryptic Soy Broth (TSB) by an inhibition assay. All S. papillosum and S. cristatum moss extracts used in this assay were prepared as described above. TSB was also prepared, autoclaved and stored at 4°C prior to use. The following equipment was used: a) Beckman® DU-64 Spectrophotometer b) Incubator Oven (commercially available from Boekel Instruments Model #133000) c) 5 mL and 50mL Polystyrene Tubes (commercially available from Falcon) d) lO ⁇ L and 1 mL polypropylene tips (commercially available from Pipetman)
• TSB Bacto® Tryptic Soy Broth
• 30 g/L commercially available from Becton Dickenson
• Staphylococcus Aureus frozen culture stock grown in TSB for a minimum of 3 log growth phases (ATCC Strain #29213 (American Type Culture Collection of Manassas, VA))
• Distilled Water commercially available from Premium Water Inc.
• TSB nutrient broth was prepared by adding 30 g/L Bacto® Tryptic Soy Broth to distilled water. The solution was stirred with a stir-bar until all the powder was dissolved and autoclaved at 121°C for 20 minutes. S. papillosum extract was prepared as described in B, above. One mL of the solutions, TSB or moss-treated TSB sample, was pipetted into 3 to 5 mL polystyrene test tubes.
• Example 4 The following data illustrate the effect of freatment of bacterial growth media (i.e., Tryptic Soy Broth) with various moss species according to the procedure in Example 3. Non-sphagnum species are very poor at preventing E. coli growth. Of the moss tested below, the most effective sphagnum mosses to prevent E. coli growth were S. papillosum and S. cristatum. "N” refers to the number of tests, “Range” presents the highest and lowest numbers obtained for these tests; and “Mean” refers to the mean value of the tests.
• N refers to the number of tests
• Range presents the highest and lowest numbers obtained for these tests
• Mean refers to the mean value of the tests.
• Example 5 TSB was treated with S. papillosum and the ability of this solution to support bacterial growth was measured according to Example 3. The percent inhibition of bacterial growth for E. coli (clinical isolate) and S. aureus (ATCC Strain # 29213) is reported. A TSB Control sample and moss-treated TSB solutions (MT-TSB) are reported below. The OD of a blank (B) is subtracted from the measured OD (Meas.) of the sample to obtain the reported Value.
• Test Material E. coli S. aureus B Meas. Value B Meas. Value
• Example 6 This example demonstrates that freatment with moss does not kill the bacteria but it does inhibit their growth.
• a fluorescence assay commercially available from Molecular Probes, Eugene, Oregon, Kit No. L-7012, was used to determine the viability of bacteria.
• This system uses mixtures of green and red fluorescent nucleic acid stains that have differing ability to penetrate viable and non-viable bacterial cells.
• the green fluorescent strain which emits at 500 nm, binds to both viable and non-viable bacteria.
• the red fluorescent strain which emits at 650 nm, binds only to non- viable bacteria. Therefore, a bacterial sample containing a higher proportion of non- viable bacteria will have an altered staining ratio.
• the data show that for both E. coli and S. aureus, the ratio of viable to non- viable bacteria remains the same as in the control sample.
• Styo-9 dye was mixed with 40 ⁇ L Propidium Iodide dye. Caution should be used as these compounds are believed to be carcinogens.
• the solutions were mixed thoroughly.
• the fluorescence intensity is measured at 500 nm (with absorption at 480 nm) and at 650 nm (with absorption at 490 nm).
• the ratio of these two values relates to the degree of viability of the bacterial culture.
• the following tables report the fluorescence intensity at two wavelengths for bacterial samples in two different media.
• the intensity of fluorescence at 500 nm over the intensity of the fluorescence at 650 nm creates a ratio which relates to the degree of viability of the bacterial culture.
• the mean of three TSB and three moss- treated TSB samples (MT-TSB) is reported below. In each case, the percent of inhibition is compared to a control sample.
• Example 7 Various bacteria were treated with S. papillosum TSB (concenfration of 50 mg/mL) according to Example 3. The percent inhibition of bacterial growth is reported. Both a TSB Confrol sample and moss-freated TSB solutions (MT-TSB) are reported below. The "Value” is obtained by subtracting the optical density (OD) of a blank (B) from the measured OD (Meas.) of a sample.
• OD optical density
• Test Material (1) E. coli S. ! aureus (clinical isolate) ATCC # 29213 B Meas. Value B Meas. Value
• Test Material (2 E. coli S. ⁇ aureus B Meas. Value B Meas. Value
• Test Material (1) S. epidermidis P. aeruginosa ATTC # 12228 ATTC #10145 B Meas. Value B Meas. Value
• Test Material (1) C. albicans A, . amsterodami ATTC # 10231 ATTC # 1001 B Meas. Value B Meas. Value
• Test Material (2 C. albicans A, . amsterodami B Meas. Value B Meas. Value
• Example 8 Effect of Acid Treatment of the Moss Compressed sticks of S. cristatum moss (obtained from Sutton's Moss, Canada (harvested in New Zealand)) were soaked in four increasing concentrations of Fe (Fe standard in concentrated HCl, 0, 0.5,5,50 mg/L, available from Ricca Chemicals, Arlington, Texas) at 50 mg moss/ml in distilled water. The soaked moss was stored overnight at 4 C. The Fe solutions were extracted by syringe, filtered and measured for Fe by inductively coupled plasma atomic emission spectrometry analysis. The results showed that the moss bound significant amounts of Fe, up to 15 mg/L in the 50 mg/L sample.
• Control 200 198.10 1.83 6.76
• Acetic Acid/Na acetate washed 500.00 307.28 7.25
• the moss was shown to bind both Fe and Ca ions, and therefore is effective in water treatment because the removal of one or both of these ions is a goal of water treatment.
• the above description and the drawings are provided for the purpose of describing embodiments of the invention and are not intended to limit the scope of the invention in any way. It will be apparent to those skilled in the art that various modifications and variations can be made without departing from the spirit or scope of the invention. Thus, it is intended that the present invention cover the modifications and variations of this invention provided they come within the scope of the appended claims and their equivalents.

Landscapes

• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Engineering & Computer Science (AREA)
• General Health & Medical Sciences (AREA)
• Organic Chemistry (AREA)
• Chemical & Material Sciences (AREA)
• General Chemical & Material Sciences (AREA)
• Medicinal Chemistry (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Pharmacology & Pharmacy (AREA)
• Animal Behavior & Ethology (AREA)
• Public Health (AREA)
• Veterinary Medicine (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Dermatology (AREA)
• Water Supply & Treatment (AREA)
• Environmental Sciences (AREA)
• Hydrology & Water Resources (AREA)
• Agronomy & Crop Science (AREA)
• Biotechnology (AREA)
• Microbiology (AREA)
• Mycology (AREA)
• Plant Pathology (AREA)
• Dentistry (AREA)
• Wood Science & Technology (AREA)
• Zoology (AREA)
• Environmental & Geological Engineering (AREA)
• Oncology (AREA)
• Communicable Diseases (AREA)
• Agricultural Chemicals And Associated Chemicals (AREA)
• Hydroponics (AREA)
• Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
• Water Treatment By Sorption (AREA)
• Cleaning By Liquid Or Steam (AREA)
• Apparatus For Disinfection Or Sterilisation (AREA)
• Medicines Containing Plant Substances (AREA)
• Pretreatment Of Seeds And Plants (AREA)
• Micro-Organisms Or Cultivation Processes Thereof (AREA)

Priority Applications (7)

Applications Claiming Priority (4)

Publications (2)

Family

ID=34966785

Family Applications (1)

Country Status (7)

Cited By (4)

Family Cites Families (8)

• 2005
  • 2005-04-14 NZ NZ551183A patent/NZ551183A/en not_active IP Right Cessation
  • 2005-04-14 CN CN2005800115381A patent/CN1942405B/zh not_active Expired - Fee Related
  • 2005-04-14 EP EP05738766A patent/EP1747174A2/en not_active Withdrawn
  • 2005-04-14 AU AU2005235588A patent/AU2005235588B2/en not_active Ceased
  • 2005-04-14 JP JP2007508581A patent/JP2007533378A/ja active Pending
  • 2005-04-14 WO PCT/US2005/012915 patent/WO2005102935A2/en active Application Filing
• 2006
  • 2006-11-13 NO NO20065204A patent/NO20065204L/no not_active Application Discontinuation

Also Published As

Similar Documents

Legal Events