Indicator device and system for detecting food spoilage and uses thereof - Patents.com

Abstract

Provided herein is an apparatus for determining the freshness of a food product and for detecting food spoilage that includes a substrate and an indicator composition. Also provided are systems including same and uses thereof.

Description

[Technical field]

[0001] Provided herein are devices, systems including same, and methods of use for determining the freshness of food products and for detecting food spoilage. [Background technology]

[0002] The spoilage of fresh food products over time is an ongoing challenge for consumers and the food product industry. Labeling of "Expiration Date", "Use Before", "Best Before" dates is often unreliable as they often do not reflect the actual quality (freshness) of the food product. It is well known that at many stages of sale and consumption (packaging or wrapping, transportation, distribution, storage, home use, etc.), the conditions necessary to maintain the freshness of food products are not always adequate, which can cause spoilage before the expected expiration date.

[0003] Moreover, expiration dates are calculated for entire food product categories for every batch (not even per food package, but per batch), a major caution factor, and therefore, by definition, they are not accurate, which results in waste of more than about 10% of the total food product production by manufacturers, retailers and final consumers. Summary of the Invention [Problem to be solved by the invention]

[0004] Thus, there is an unmet need for indicator devices and systems that are capable of providing a continuous, real-time indication of food freshness or spoilage or the presence of pathogens for each food package, and that are cost-effective and easy to implement on or with existing food packaging. [Means for solving the problem]

[0005] Disclosed herein is an indicator device configured to provide a real-time indication of food freshness, food spoilage and / or the presence of pathogens. Advantageously, the device is configured to allow unidirectional contact between food products or reactants secreted therefrom and an indicator agent (indicator composition), while preventing / avoiding or allowing only minor movement of the indicator agent into the food product. The unidirectional contact may be momentary, i.e., a one-time contact, or continuous contact. Nevertheless, the indication remains informative and reliable in any one of the aforementioned configurations.

[0006] According to some embodiments, the indicator device may be in continuous contact with the food product or in continuous contact with a portion thereof followed by a short transient passage of said food product, a portion thereof, or a component secreted therefrom through the indicator device. Thus, contact between the food product and the indicator composition / device may be automatic or may be manually enabled, i.e., following the opening of a passage between the indicator device and the food product, contact between the food product and the indicator composition / device. According to some embodiments, contact may be direct or indirect. In some embodiments, contact is a fluid (gas or liquid) contact. In some embodiments, the device may be directly placed / associated with the food product and contact between the food product and the indicator portion of the device may be automatically / spontaneously facilitated.

[0007] In some embodiments, the indicator device is disposed / positioned (permanently or temporarily) on a food container / package and configured to allow the food product, a portion thereof, or a component secreted therefrom to pass temporarily, unidirectionally through the indicator device to provide an indication regarding the food product therein.

[0008] In some embodiments, the indicator device is configured to be placed / positioned (permanently or temporarily) directly on the food product and to allow components present on / with the food product or secreted from the food product to pass temporarily (preferably in one direction) through the indicator device to provide an indication regarding the food product.

[0009] According to some embodiments, the indicator device or indicator composition may be placed within or in direct contact with the food product and a "spoilage signal" (e.g., a color change reaction, a spectroscopic indication, a colorimetric indication, a fluorometric indication, an electrochemical indication, a phase transition (such as phase separation), precipitation or gas evolution) may be conveyed / transmitted to the food surface or exterior of the food package. Such transmission of an internal spoilage signal (e.g., a color change) may be accomplished, for example, by mirror reflection, a magnifying glass, an electronic device, or any other suitable means.

[0010] In some embodiments, the device indicators are configured to be placed / positioned / present on individual or multiple food containers, and the food products in said containers may be any type of food product having any type of shape, form, or composition, including, for example, solid, semi-solid, semi-liquid, liquid, frozen, thawed, etc. forms.

[0011] According to some embodiments there is provided an indicator device for detecting food spoilage in a food product, the indicator device comprising: a. Substrate and b. an indicator composition comprising at least one indicator or indicator, said indicator composition being bound to said substrate, said substrate being configured to contact a food product, and said indicator composition being configured to generate a detectable signal upon reaction with at least a portion of or a component secreted therefrom of the food product.

[0012] According to some embodiments, the substrate may include or be made from textile fibers, plastics, nylon, woven fibers, woven fabrics, nonwoven fabrics, man-made polymers, biodegradable polymers, cotton, wool, linen, oligosaccharides, gels, modified starches, metals, aluminum, paper, carton, wood, plant materials, clay, adhesives, ointments, vax, oils, leather, carbon and graphite felt, carbon fibers, or any combination thereof.

[0013] According to some embodiments, the substrate may include or be made from woven fibers, plastic, nylon, woven fibers, nonwoven fibers, cotton, silk, wool, filter paper (such as coffee filters), or any combination thereof. In some embodiments, the substrate may further include a coating layer to facilitate placement / attachment to a food product or food packaging.

[0014] According to some embodiments, the substrate may further include one or more additional layers, in some embodiments, at least one of the layers is configured to at least partially prevent leakage of the indicator composition into the food product.

[0015] According to some embodiments, the additional layer may include or be made of textile fibers, plastics, nylon, woven fibers, nonwoven fibers, cotton, wool, silk, filter paper, cellulose-starch chitosan-PVA, cellulose-PVA crosslinked films, or any combination thereof. In some embodiments, the substrate and / or additional layer may be treated with various substances to enhance binding with various substances and / or indicator compositions. Indicator compositions include, but are not limited to, substances such as, for example, non-organic salts, organic cationic salts, proteins (e.g., food approved), etc., or any combination thereof.

[0016] According to some embodiments, the additional layer may be permanently or temporarily associated with the substrate, hi some embodiments, the additional layer may be stitched, adhered, glued, and / or welded to the substrate.

[0017] According to some embodiments, spoilage is microbial-associated spoilage, including any type of microorganism, including, for example, but not limited to, bacteria, fungi, molds, pathogens, and the like.

[0018] According to some embodiments, the at least one indicator is a microbial indicator, an azo dye, a food color, a natural food dye, a thiazine dye, a redox indicator, a pH indicator, an oxygen indicator, a solvatochromic dye, a nanobiosensor (e.g., gold nanoparticles, silver nanoparticles, quantum dots, magnetic nanoparticles, single and / or multi-walled nanotubes, graphene and reduced graphene flakes, TiO 2 The biosensor may be selected from a nanoparticle, a fluorescent polymer nanoparticle, a nanotip, a nanowire, etc.), a fiber optic biosensor, or any combination thereof. Each possibility is a separate embodiment.

[0019] According to some embodiments, the at least one indicator may be provided at a concentration that is detectable (e.g., a change in color, spectroscopic, colorimetric, fluorometric, electrochemical change) in response to spoilage of the food sample (e.g., a visual change). According to some embodiments, the at least one indicator may be a pH indicator, and the indicator may change color upon a change in pH of less than about 0.1 pH units. According to some embodiments, the at least one indicator may be a pH indicator, and the indicator may change color upon a change in pH of less than about 0.5 pH units. According to some embodiments, the at least one indicator may be a bacterial indicator, and may be free of quaternary ammonium salts and quaternary amine moieties. According to some embodiments, the at least one bacterial indicator may provide a detectable indication in the presence of a bacterial population above a predetermined threshold. According to some embodiments, the at least one bacterial indicator may provide a detectable indication in the presence of a volatile compound secreted from a food product above a threshold, for example, due to a bacterial population above a predetermined threshold. For example, the indicator composition may include ninhydrin, phthalocyanine and / or azo dyes, which form colored complexes with volatile amines that are products of bacterial decomposition of proteins (biogenic amines), such as putrescine, ammonia, urea and others.

[0020] According to some embodiments, the at least one indicator may be selected from bromothymol blue, cresol red, phenol red, methyl red, bromocresol blue, indigo carmine, carmoisine red, tartrazine, bromocresol green, methyl orange, brown HT, methyl blue, sunset yellow FCF, citrus red 2, amaranth, ponceau 4R, quinoline yellow, carmoisine, patent blue V, green S, brilliant blue, indigotine, fast green FCF, methylene blue, erythosine, allura red AC, orange B, scarlet GN, sudan II, acid orange 20, annatto, caramel color, carmine, elderberry, lycopene, paprika, turmeric, tetrazolium salts, resazurin, ninhydrin, phthalocyanines, azo dyes, or any combination thereof.

[0021] According to some embodiments, the indicator composition may include multiple indicators.

[0022] According to some embodiments, the indicator composition may be linked to the substrate. In some embodiments, the indicator composition is essentially irreversibly linked to the substrate. In some embodiments, the indicator composition may be reversibly linked to the substrate. In some embodiments, the indicator composition may be covalently bonded to the substrate.

[0023] According to some embodiments, the substrate may have associated therewith a plurality of indicator compositions. According to some embodiments, the plurality of indicator compositions may be spatially distinct.

[0024] According to some embodiments, the multiple indicator compositions may differ with respect to indicator type and / or indicator concentration.

[0025] According to some embodiments, the substrate may be in fluid (such as gas or liquid) contact with the food sample. In some embodiments, the substrate may be configured to directly or indirectly contact the food product.

[0026] According to some embodiments, the device may be in the form of a patch, sticker, flag, note, or the like.

[0027] According to some embodiments, the detectable signal may be qualitative or quantitative, hi some embodiments, the detectable signal is colorimetric, spectroscopic, fluorometric, and / or electrochemical.

[0028] According to some embodiments, the indicator device may be or may be included with a food package configured to at least partially hold or encase a food product.

[0029] According to some embodiments, there is provided a method for detecting food spoilage of a food product, the method comprising the steps of placing an indicator device as disclosed herein in contact with a food product, contacting at least a food sample of the food product with the indicator, and determining whether the food product has spoiled upon detecting a detectable signal produced by the indicator if / when the food sample has spoiled.

[0030] According to some embodiments, a system for detecting food spoilage of a food product is provided, the system including at least one food package configured to at least partially hold or encase a food product, and an indicator device as disclosed herein coupled to the food package.

[0031] According to some embodiments there is provided an indicator device configured to couple with a food package (capable of holding a food product) and to indicate spoilage of a food product contained in the food package, the indicator device comprising: (a) a substrate comprising an indicator composition, the indicator composition comprising at least one indicator; and (b) at least one passageway configured to permit unidirectional passage of a food sample (from a food package) into the indicator device; The indicator composition is configured to produce a detectable signal upon reaction with a food sample, the signal being indicative of food spoilage.

[0032] According to some embodiments, the food sample may comprise a portion of a food product and / or components secreted therefrom.

[0033] According to some embodiments, the indicator composition may be associated with a substrate.

[0034] According to some embodiments, the spoilage is microbial-associated spoilage.

[0035] According to some embodiments, the at least one indicator may be selected from a microbial indicator, an azo dye, a food dye, a natural food dye, a thiazine dye, a redox indicator, a pH indicator, an oxygen indicator, a solvatochromic dye, or any combination thereof.

[0036] According to some embodiments, the substrate may include or be made from textile fibers, plastics, nylon, woven fibers, fabrics, nonwovens, man-made polymers, biodegradable polymers, cotton, wool, linen, oligosaccharides, gels, modified starches, metals, aluminum, paper (e.g., coffee filters), carton, wood, plant materials, clay, adhesives, ointments, buckwheat, oils, leather, silk, or any combination thereof. Each possibility is a separate embodiment.

[0037] According to some embodiments, the at least one indicator may be provided at a concentration that promotes a detectable signal (e.g., a color change) in response to spoilage of the food sample. According to some embodiments, the at least one indicator may be a pH indicator, and the indicator may change color upon a change in pH of less than about 0.1 pH units. According to some embodiments, the at least one indicator may be a pH indicator, and the indicator may change color upon a change in pH of less than about 0.5 pH units. According to some embodiments, the at least one indicator may be a bacterial indicator, and may be free of quaternary ammonium salts and quaternary amine moieties. According to some embodiments, the at least one bacterial indicator may provide a detectable indication in the presence of a bacterial population above a predetermined threshold.

[0038] According to some embodiments, the at least one indicator may be selected from bromothymol blue, cresol red, phenol red, methyl red, bromocresol blue, indigo carmine, carmoisine red, tartrazine, bromocresol green, methyl orange, methyl blue, sunset yellow FCF, citrus red 2, amaranth, ponceau 4R, quinoline yellow, carmoisine, patent blue V, green S, brilliant blue, indigotine, fast green FCF, erythrocin, allura red AC, orange B, scarlet GN, sudan II, acid orange 20, annatto, caramel color, carmine, elderberry, lycopene, paprika, turmeric, tetrazolium salts, resazurin, ninhydrin, phthalocyanines, azo dyes, or any combination thereof.

[0039] According to some embodiments, the indicator composition may include multiple indicators.

[0040] According to some embodiments, the indicator composition may be linked to the substrate. In some embodiments, the indicator composition is essentially irreversibly linked to the substrate. In some embodiments, the indicator composition may be reversibly linked to the substrate. In some embodiments, the indicator composition may be covalently bonded to the substrate.

[0041] According to some embodiments, at least one passageway may be a closed passageway configured to be temporarily opened when required.

[0042] According to some embodiments, the at least one passage may include at least one capillary, a membrane, a displacement element, or any combination thereof. According to some embodiments, the displacement may be a rotation or a press.

[0043] According to some embodiments, at least one passageway may be configured to open one or more times.

[0044] According to some embodiments, the indicator composition may be linked to the substrate. In some embodiments, the indicator composition is essentially irreversibly linked to the substrate. In some embodiments, the indicator composition may be reversibly linked to the substrate. In some embodiments, the indicator composition may be covalently bonded to the substrate.

[0045] According to some embodiments, the substrate may have associated therewith a plurality of indicator compositions. According to some embodiments, the plurality of indicator compositions may be spatially distinct.

[0046] According to some embodiments, the multiple indicator compositions may differ with respect to indicator type and / or indicator concentration.

[0047] According to some embodiments, the substrate may be in fluid (such as gas or liquid) contact with the food sample. In some embodiments, the substrate may be configured to directly or indirectly contact the food product.

[0048] According to some embodiments, the substrate may be in fluid (such as gas or liquid) contact with the food sample. In some embodiments, the substrate may be configured to directly or indirectly contact the food product.

[0049] According to some embodiments, the detectable signal may be qualitative or quantitative. In some embodiments, the detectable signal may be colorimetric.

[0050] According to some embodiments, there is provided a method for detecting food spoilage of a food product contained in a food package, the method comprising the steps of coupling an indicator device in contact with the packaged food product; temporarily allowing passage of a food sample in one direction from the food package towards the indicator device by opening a closed passageway, thereby allowing the food sample to contact and react with the indicator composition; and upon detecting a detectable signal, determining whether the food product has spoiled.

[0051] According to some embodiments, there is provided a system for detecting food spoilage of a food product, the system including at least one food package configured to hold a food product, and an indicator device as disclosed herein (including at least one passageway) associated with the food package.

[0052] According to some embodiments, the food product may be selected from a pourable food product, a liquid food product, a soft food product, a semi-rigid food product, a rigid food product and an aqueous food product.

[0053] According to some embodiments, the food packaging may be selected from containers, vials, boxes, trays, cans, and pallets, bags, wraps, nylon wraps, Saran Wrap, smart packages, or any combination thereof.

[0054] According to some embodiments, the indicator device may be integrally formed with the food package. According to some embodiments, the indicator device may be permanently associated with the food package. According to some embodiments, the indicator device may be temporarily associated with the food package. According to some embodiments, the indicator device may be associated with an exterior region of the food package.

[0055] Other objects, features and advantages of the present invention will become apparent from the following description, examples and drawings.

[0056] Certain embodiments of the present disclosure may include some, all, or any of the advantages described above. One or more other technical advantages will be readily apparent to those skilled in the art from the drawings, descriptions, and claims contained herein. Moreover, although certain advantages have been enumerated above, various embodiments may include all, some, or none of the enumerated advantages. [Brief description of the drawings]

[0057] Some embodiments of the present disclosure are described herein with reference to the accompanying drawings. The description, together with the drawings, will make clear to those skilled in the art how some embodiments can be implemented. The drawings are for illustrative purposes and are not intended to show structural details of the embodiments in more detail than is necessary for a basic understanding of the present disclosure. For clarity, some objects shown in the figures are not drawn to scale.

[0058] [Figure 1A]Figure 1A shows a pictogram of four vials, each containing whole milk and wool stained with 50 ppm indigo carmine solution (i.e., wool stained with indigo carmine solution at a concentration of 50 ppm), immediately after incubation at room temperature (25–27 °C) at t = 0 (Figure 1A). [Figure 1B] Figure 1B shows a pictogram of four vials, each containing whole milk and wool stained with 50 ppm indigo carmine solution (i.e., wool stained with an indigo carmine solution at a concentration of 50 ppm), incubated at room temperature (25-27 °C) and showing their state after approximately 30 hours (Figure 1B). [Figure 1C] Figure 1C shows a pictogram of four vials, each containing whole milk and wool stained with 50 ppm indigo carmine solution (i.e., wool stained with an indigo carmine solution at a concentration of 50 ppm), after approximately 48 hours of incubation at room temperature (25–27 °C) (Figure 1C). [Figure 2A] Figure 2A shows a fresh sample of milk containing wool fragments stained with 60 ppm indigo carmine solution prior to incubation at room temperature (25-27 °C) for 36-40 hours. [Figure 2B] Figure 2B shows a fresh sample of milk containing wool fragments stained with 60 ppm indigo carmine solution after 36-40 hours of incubation at room temperature (25-27 °C). [Figure 3A] FIG. 3A shows at t=0 a vial containing chicken fillets encased within cotton fabric dyed with a neutral or basic cationic solution. [Figure 3B] FIG. 3B shows at t=0 a vial containing red meat encased within a cotton fabric dyed with a neutral or basic cationic solution. [Figure 3C] FIG. 3C shows the vial shown in FIG. 3A after incubation at room temperature (26-28° C.) for 30-36 hours. [Figure 3D] FIG. 3D shows the vial shown in FIG. 3B after incubation at room temperature (26-28° C.) for 30-36 hours. [Figure 4A] FIG. 4A shows the state at t=0 of a vial containing cottage cheese encased within cotton fabric dyed with a neutral cationic solution. [Figure 4B] FIG. 4B shows the condition at t=28 hours of a vial containing cottage cheese encased within cotton fabric dyed with a neutral cationic solution. [Figure 5A] FIG. 5A shows cotton fabric dyed with a neutral cationic solution at t=0 in a vial containing soy milk at room temperature (26-28° C.). [Figure 5B] FIG. 5B shows cotton fabric dyed with a neutral cationic solution at room temperature (26-28° C.) in a vial containing soy milk for t=28-30 h. [Figure 6A] Figure 6A shows pictograms of vials containing cotton fabric that binds the indigo carmine indicator incubated at room temperature (26-28 °C) in the presence of cottage cheese, at t = 0 (Figure 6A). Each pictogram shows a vial containing only a food sample (cottage cheese), a vial containing only cotton fabric that binds the indigo carmine indicator, and a vial containing both cotton fabric and a food sample that binds the indigo carmine indicator. [Figure 6B] Figure 6B shows pictograms of vials containing cotton fabric that binds the indigo carmine indicator incubated at room temperature (26-28 °C) in the presence of cottage cheese, at t = 24 (Figure 6B). Each pictogram shows a vial containing only a food sample (cottage cheese), a vial containing only cotton fabric that binds the indigo carmine indicator, and a vial containing both cotton fabric and a food sample that binds the indigo carmine indicator. [Figure 6C] Figure 6C shows pictograms of vials containing cotton cloth bound to the indigo carmine indicator incubated at room temperature (26-28 °C) in the presence of minced chicken breast meat, at t = 0 (Figure 6C). Each pictogram shows a vial containing only the food sample (minced chicken breast meat), a vial containing only the cotton cloth bound to the indigo carmine indicator, and a vial containing both the cotton cloth and the food sample bound to the indigo carmine indicator. [Figure 6D] Figure 6D shows pictograms of vials containing cotton cloth bound to the indigo carmine indicator incubated at room temperature (26-28 °C) in the presence of minced chicken breast meat, at t = 24 (Figure 6D). Each pictogram shows a vial containing only a food sample (minced chicken breast meat), a vial containing only cotton cloth bound to the indigo carmine indicator, and a vial containing both cotton cloth and a food sample bound to the indigo carmine indicator. [Figure 7A] Figure 7A presents a pictogram of fresh fish meat (salmon) with an indicator device coupled to a meat package (packaging in the form of a nylon wrap). Fresh meat (t=0) is shown in Figure 7A. A color change in indicator for fresh meat (Figure 7A) and spoiled meat (Figure 7B) is visible. [Figure 7B] Figure 7B presents a pictogram of fresh fish meat (salmon) with an indicator device coupled with the meat package (packaging in the form of nylon wrapping). In Figure 7B spoiled meat (after 24-36 hours at room temperature) is shown. A color change of the indicator is visible for fresh meat (Figure 7A) and spoiled meat (Figure 7B). [Figure 8A] 8A shows a perspective view of an indicator device, according to some embodiments. [Figure 8B] FIG. 8B shows pictograms and corresponding diagrams of layers / portions of the indicator device. [Figure 8C] FIG. 8C shows a pictogram of the indicator device and its layers coupled to a food product. [Figure 9] FIG. 9 shows a perspective view of an indicator device that allows unidirectional food sample passage, according to some embodiments. [Figure 10A]Figures 10A-10E show perspective views of an indicator device and a system including the same, according to some embodiments. The device shown in Figures 10A-D shows pressure-initiated unidirectional food sample passage, whereby the passage is opened by pushing on the device, thereby allowing the food sample to enter the indicator. When pressure is released, the passage is sealed / closed. Figure 10A shows a perspective view of the indicator device (Figure 10A) without the outer cover. [Figure 10B] Figures 10A-10E show perspective views of an indicator device and a system including the same, according to some embodiments. The device shown in Figures 10A-D shows pressure-initiated unidirectional food sample passage, whereby the passage is opened by pushing on the device, thereby allowing the food sample to enter the indicator. When pressure is released, the passage is sealed / closed. Figure 10B shows a perspective view of the indicator device (Figure 10B) with an outer cover. [Figure 10C] Figures 10A-10E show perspective views of an indicator device and a system including the same, according to some embodiments. The device shown in Figures 10A-D shows pressure initiated unidirectional food sample passage, whereby the passage is opened by pressing the device, thereby allowing the food sample to enter the indicator. When pressure is released, the passage is sealed / closed. Figure 10C shows a perspective view of a system including a food package coupled to an indicator device. [Figure 10D] Figures 10A-10E show perspective views of an indicator device and a system including the same, according to some embodiments. The device shown in Figures 10A-D shows a pressure initiated, one-way food sample passageway whereby the passageway is opened by pressing the device, thereby allowing the food sample to enter the indicator. When pressure is released, the passageway is sealed / closed. Figure 10D shows a close-up view of a cross section of the device mating with a food package having a closed passageway (Figure 10D). [Figure 10E]Figures 10A-10E show perspective views of an indicator device and a system including the same, according to some embodiments. The device shown in Figures 10A-D shows a pressure initiated, one-way food sample passageway whereby the passageway is opened by pressing the device, thereby allowing the food sample to enter the indicator. When pressure is released, the passageway is sealed / closed. Figure 10E shows a close-up view of a cross section of the device mating with a food package having an open passageway (Figure 10E). [Figure 11A] 11A-B show perspective views of an indicator device and a system including the same, according to some embodiments. The device and system shown in FIG. 11A-B show pressure initiated unidirectional food sample passage, whereby pressing on the device opens the passage, thereby allowing the food sample to enter the indicator. When pressure is released, the passage is sealed / closed. FIG. 11A shows a perspective view of a system including an indicator device and a food package. [Figure 11B] 11A-B show perspective views of an indicator device and a system including the same, according to some embodiments. The device and system shown in FIG. 11A-B show a pressure initiated one-way food sample passageway whereby pressing on the device opens the passageway, thereby allowing the food sample to enter the indicator. When pressure is released, the passageway is sealed / closed. FIG. 11B shows a close up view of a cross section of the device coupling to a food package at the closed passageway and the open passageway. [Figure 12] 12 shows a perspective view of an indicator device and a system including the same, according to some embodiments. The device and system shown in FIG. 12 shows pressure initiated unidirectional food sample passage, whereby passage is opened by pressing the device, opening an internal opening in the food package, thereby allowing the food sample to enter the indicator. When pressure is released, the passage is sealed / closed. [Figure 13A]13A-B show perspective views of an indicator device and a system including the same, according to some embodiments. The device and system shown in FIG. 13A-B show unidirectional food sample passage initiated by capillary forces, which is facilitated by a capillary tube that fluidly connects the food product and the indicator device. Once the indicator is filled (or saturated) with the food sample, there is no further fluid flow between the indicator device and the food product. FIG. 13A shows a perspective view of a system including an indicator device that couples to a food package. [Figure 13B] 13A-B show perspective views of an indicator device and a system including the same, according to some embodiments. The device and system shown in FIG. 13A-B show unidirectional food sample passage initiated by capillary forces, which is facilitated by a capillary tube that fluidly connects the food product and the indicator device. Once the indicator is filled (or saturated) with food sample, there is no further fluid flow between the indicator device and the food product. FIG. 13B shows a cross-sectional view of an inner wall of a food package of the system including a capillary passage system, facilitating unidirectional passage of food sample from the food package to the indicator device. [Figure 14A] 14A-14B show perspective views of an indicator device and a system including the same, according to some embodiments. The device and system shown in FIGS. 14A-14B show one-way food sample passage facilitated by a valve unit, whereby one valve is open and one valve is closed, and by turning / rotating an actuation button, the open valve is closed and the closed valve is opened. Such a mechanism allows for stepwise passage of food sample in one direction (unidirectional) from the food container to the indicator device. Once the food sample has passed, the indicator device may be locked for further passage, if desired. FIG. 14A shows a cross-sectional internal view of a food package of the system, which couples to the indicator device. [Figure 14B]14A-14B show perspective views of an indicator device and a system including the same, according to some embodiments. The device and system shown in Figs. 14A-14B show one-way food sample passage facilitated by a valve unit, whereby one valve is open and one valve is closed, and by turning / rotating an actuation button, the open valve is closed and the closed valve is opened. Such a mechanism allows for stepwise passage of food sample in one direction (unidirectional) from the food container to the indicator device. Once the food sample has passed, the indicator device may be locked for further passage, if desired. Fig. 14B shows the passage valve unit in different configurations (open and closed). [Figure 15A] 15A-15C show perspective views of an indicator device and a system including the same, according to some embodiments. The devices and systems shown in Figures 15A-C show a one-way food sample passing from a solid / semi-solid food product to the indicator device. Figure 15A shows an exemplary indicator device configured to be placed / positioned on a food package. [Figure 15B] Figures 15A-15C show perspective views of an indicator device and a system including the same, according to some embodiments. The devices and systems shown in Figures 15A-C show a one-way food sample passing from a solid / semi-solid food product to the indicator device. Shown in Figure 15B is an indicator device placed on a solid food product. The indicator device includes an area / chamber where the food product or a sample from the food product passes through a dedicated tube / channel / capillary. [Figure 15C] Figures 15A-15C show perspective views of an indicator device and a system including the same, according to some embodiments. The devices and systems shown in Figures 15A-C show a one-way food sample passing from a solid / semi-solid food product to the indicator device. Shown in Figure 15C is an indicator device placed on a solid food product. The indicator device includes an area / chamber where the food product or a sample from the food product passes through a dedicated tube / channel / capillary. [Figure 16A]Figure 16A is a pictogram of a fresh (Figure 16A) chicken breast food sample binding to an indicator device, showing a color change in the indicator device at the indicated colony forming unit (CFU) count. [Figure 16B] Figure 16B is a pictogram of an autoclaved (Figure 16B) chicken breast food sample binding to an indicator device showing a color change in the indicator device at the indicated colony forming unit (CFU) count. [Figure 17A] Figures 17A-D show pictograms of various regions of fresh chicken breast sections and the corresponding microbial counts stored for 5 days at 4° C. Figure 17A is a pictogram of a chicken breast section (showing the anterior, apical, and posterior regions). [Figure 17B] Figures 17A-D show pictograms and corresponding microbial counts for various regions of fresh chicken breast sections stored for 5 days at 4° C. Figure 17B is a bar graph showing the microbial counts (log CFU / gr) for various chicken breast regions. [Figure 17C] Figures 17A-D show pictograms of various regions and corresponding microbial counts of fresh chicken breast sections stored for 5 days at 4° C. Figure 17C shows pictograms showing the inner (internal) or upper (external) regions. [Figure 17D] Figures 17A-D show pictograms and corresponding microbial counts of various regions of fresh chicken breast sections stored for 5 days at 4° C. Figure 17D is a bar graph showing microbial counts (log CFU / gr) of chicken breast regions. [Figure 18A] Figures 18A-D show pictograms of chicken breast samples packaged in various types of packaging and binding to the indicator device before and after incubation (4°C, 2 days). Additionally, the test CFU (log / gr) and coefficient of variation (CV) percentages are shown when indicator discoloration is observed. Figure 18A shows chicken breast packaged in polyethylene wrap and binding to an indicator device containing an indicator composition of 10 ppm indigo carmine and a mediator layer. As shown in Figures 18A-D, colored indicators ("fresh" samples) and discolored indicators ("spoiled" samples) are shown. [Figure 18B] Figure 18A-D shows pictograms of chicken breast samples packaged in various types of packaging and binding to the indicator device before and after incubation (4°C, 2 days). Additionally, the test CFU (log / gr) and the coefficient of variation (CV) percentage when indicator discoloration is observed are shown. Figure 18B shows chicken breast packaged in a vacuum pack and binding to an indicator device containing an indicator composition of 10 ppm indigo carmine and a mediator layer. As shown in Figure 18A-D, colored indicators ("fresh" samples) and discolored indicators ("spoiled" samples) are shown. [Figure 18C] Figure 18A-D shows pictograms of chicken breast samples packaged in various types of packaging and binding to the indicator device before and after incubation (4°C, 2 days). Additionally, the test CFU (log / gr) and coefficient of variation (CV) percentages are shown when indicator discoloration is observed. Figure 18C shows chicken breast packaged in a vacuum pack and binding to an indicator device containing an indicator composition of 10 ppm indigo carmine and a mediator layer. As shown in Figure 18A-D, colored indicators ("fresh" samples) and discolored indicators ("spoiled" samples) are shown. [Figure 18D] Figure 18A-D shows pictograms of chicken breast samples packaged in various types of packaging and binding to the indicator device before and after incubation (4°C, 2 days). Additionally, the test CFU (log / gr) and coefficient of variation (CV) percentages are shown when indicator discoloration is observed. Figure 18D shows chicken breast packaged in a vacuum pack and binding to an indicator device containing an indicator composition of 10 ppm indigo carmine and 5 ppm tartrazine and a mediator layer. As shown in Figure 18A-D, colored indicators ("fresh" samples) and discolored indicators ("spoiled" samples) are shown. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0059] The principles, uses, and implementations of the teachings herein may be better understood with reference to the accompanying description and drawings. Upon review of the description and drawings herein, one skilled in the art will be able to practice the teachings herein without undue effort or experimentation. In the drawings, like reference numbers refer to like parts throughout. In the drawings, like reference numbers refer to like parts throughout.

[0060] In some embodiments, there is provided an indicator device attached to or coupled to a food product and configured to detect spoilage of the food product, said indicator device comprising: (a) a substrate; (b) an indicator composition that is attached to the substrate; The substrate is configured to at least partially contact the food product and the indicator composition is configured to generate a detectable signal upon reaction with a portion of the food product or with a component secreted therefrom, whereby the detectable signal is indicative of food spoilage.

[0061] In some embodiments, the substrate is non-permeable. In some embodiments, the substrate is permeable. In some embodiments, the substrate is semi-permeable. In some embodiments, one side of the substrate is permeable and the opposing (other) side is not permeable. In some embodiments, the substrate is at least partially permeable. In some embodiments, the substrate is permeable. In some embodiments, the substrate is woven. In some embodiments, the substrate is nonwoven. In some embodiments, the substrate is rigid. In some embodiments, the substrate is semi-rigid. In some embodiments, the substrate is flexible. In some embodiments, the substrate may be or may be made of any type of fibrous material that allows for the passage or absorption of fluid. In some embodiments, the substrate may comprise textile fibers. In some embodiments, the substrate may comprise naturally produced textile fibers. In some embodiments, the fibers may comprise cellulose. In some embodiments, the substrate may comprise cotton. In some embodiments, the substrate may be made from cotton. In some embodiments, the substrate may be made from plastic, polypropylene, polyethylene, a combination of plastic and fiber, or any other suitable material. In some embodiments, the substrate may be made from polypropylene nonwoven. In some embodiments, the substrate is woven. In some embodiments, the substrate may be made of or include textile fibers, plastics, nylon, woven fabrics, textiles, nonwoven fabrics, man-made polymers, biodegradable polymers, cotton, wool, linen, oligosaccharides, polysaccharides, gels, modified starches, metals, aluminum, paper, carton, wood, plant materials, clay, silk, adhesives, ointments, bags, filter paper, oils, leather, cellulose / starch chitosan / PVA (optionally bonded to fabrics), cellulose / PVA crosslinked films (optionally bonded to fabrics), carbon and graphite felt, carbon fibers, acetic acid, glycerin, or any combination thereof. Each possibility is a separate embodiment.In some embodiments, the substrate may comprise a combination of materials such as, for example, cotton treated / coated with starch (e.g., starch chitosan), film coated cotton, acetate coating, glycerin coating, PVA coating, etc., or any combination thereof.

[0062] In some embodiments, the substrate may be further coated with an additional layer (e.g., a film layer) that may be made from, for example, PVA (polyvinyl alcohol), acetic acid, glycerin, lecithin, chitosan, etc., or any combination thereof. In some embodiments, such a coating layer may at least partially prevent or reduce leakage of the instruction from the substrate into the food product.

[0063] In some embodiments, the substrate may include one or more additional layers, which may be made of, for example, but are not limited to, plastic, polypropylene, polyethylene, a combination of plastic and fiber, polypropylene nonwoven, textile material, textile fiber, plastic, nylon, woven fiber, woven fabric, nonwoven fabric, artificial polymer, biodegradable polymer, cotton, wool, linen, oligosaccharide, polysaccharide, gel, modified starch, metal, aluminum, paper, carton, wood, plant material, clay, silk, adhesive, ointment, bagus, filter paper, oil, leather, cellulose / starch chitosan / PVA (optionally bonded to fabric), cellulose / PVA crosslinked film (optionally bonded to fabric), carbon and graphite felt, carbon fiber, or any combination thereof. Each possibility is a separate embodiment. In some embodiments, the one or more additional layers may further include or be treated with cationic agent and / or NaOH. In some embodiments, the additional layer is a mediator layer. In some embodiments, the one or more additional layers are configured to collect / absorb / hold residual indicators leaking toward the food product when the residual indicators leak from the indicator composition bound to the substrate toward the food product. In some embodiments, at least one of the additional layers is disposed between the food product / food package and the substrate. In some embodiments, the one or more additional layers may be temporarily, reversibly, or permanently bound to the substrate. In some embodiments, the additional layer may include a top cover / patch configured to facilitate the placement / binding of the device and the food product / food package. In some embodiments, the binding between the substrate and the one or more additional layers may be facilitated by any means known in the art, including, for example, suturing, bonding, gluing, welding, and the like. Each possibility is a separate embodiment.

[0064] Reference is now made to FIG. 8A-B, which shows exemplary diagrams of indicator devices, according to some embodiments. Shown in FIG. 8A is an exemplary indicator device 8, which may be essentially flat, for example in the form of a sticker or patch. The indicator device 8 includes a substrate 9 that includes / is attached to / is connected to an indicator composition 10. Now referring to FIG. 8B, FIG. 8B shows a pictogram of the exemplary indicator device 12, a diagram of various parts / portions / layers, including a substrate 14 that is attached to an indicator composition 16, and an optional additional layer 13 (also called an intermediary layer) that is attached (e.g., by gluing, bonding, sewing, welding) to the substrate. The additional layer 13 is configured to at least partially prevent leakage of the indicator composition into the food product. In some embodiments, the additional layer 13 is disposed between the food product / food package and the substrate. Additionally, a top layer / cover 15 is shown, which is configured, for example, to facilitate positioning / bonding (e.g., by sticking / adhering) of the device and the food product / food package. Reference is now made to Figure 8C, which shows a pictogram of indicator device 12' disposed on a food product (chicken breast 19). The right panel shows the separate layers / portions of indicator device 12', which includes intermediate layer 13', substrate 14' which bonds to indicator composition 16', and top layer / cover 15', shown in the form of a sticker and configured to adhere / place / secure the device to the food product.

[0065] As used herein, "cotton" generally refers to a natural polymer derived from the cotton plant and is made primarily of cellulose. Cotton has the ability to absorb liquids.

[0066] In some exemplary embodiments, the indicator composition may include at least one indicator and at least one additional agent, such as, but not limited to, Therafast GMX, Denitex BC, glycidyl trimethyl ammonium chloride, betaine, casein, choline chloride, lecithin, chitosan, dicyandiamide, melamine, urea, biotin, casein, betaine chloride, proteins (such as albumin and casein), or any combination thereof. Each possibility is a separate embodiment. In some embodiments, the additive is a cationic agent. In some embodiments, some of the agents may be used as adjuvants to facilitate binding of other additives to the substrate. For example, in some embodiments, materials such as dicyandiamide, melamine, citric acid, phosphoric acid, sodium carbonate, sodium bicarbonate, sodium hypophosphite, polyvinylpyrrolidone, aluminosilicate clay, urea, etc. may be used as adjuvants, pH adjusters, and / or catalysts during the preparation / manufacture of the device. In some embodiments, the indicator composition does not contain a quaternary salt such as 2,3 epoxypropyltrimethylammonium chloride. In some embodiments, such salts are used to pretreat the cotton or cellulose and substrate, and free residues of the quaternary salt are thoroughly washed prior to the binding reaction with the indicator. In some embodiments, the indicator composition may have a pH greater than 7.

[0067] According to some embodiments, the manufacture / preparation / fabrication of the device may include one or more steps including, for example, pre-treating the substrate (e.g., with a cationic agent), coating the substrate (with one or more suitable coating solutions), binding / linking / binding the indicator composition to the substrate, binding one or more additional layers, drying, sterilizing (e.g., by autoclaving), etc., or any combination of those steps. The type, order, and length of steps of such methods are determined based on the type of device, the type of substrate, the type of indicator in the composition, etc.

[0068] According to some embodiments, sterilization of the indicator device may include any suitable sterilization method, including, for example, low temperature pasteurization (using an electron beam accelerator, E-beam), high pressure processing (HPP), 70% ethanol immersion, treatment with ethanol or ethanol-water vapor, ultraviolet light treatment, infrared light treatment, ozone treatment (e.g., by immersion in ozonated water), high temperature sterilization (e.g., at 300° C. for a very short time (minutes)), ultrasonic treatment, etc., or any combination thereof. Each possibility is a separate embodiment.

[0069] In some embodiments, the substrate may include cellulose, and the indicator composition may include at least one indicator and at least one cationic agent. In some embodiments, the cationic agent may include a quaternary ammonium compound. In some embodiments, the cationic agent may include 2,3-epoxypropyltrimethylammonium chloride. In some embodiments, the permeable substrate may include cellulose, and the indicator composition may have a pH greater than 7. In some embodiments, the substrate is pretreated with 2,3-epoxypropyltrimethylammonium chloride and a cationic agent such as NaOH, and then stained / bonded with the indicator composition (e.g., indigo carmine).

[0070] In some embodiments, the substrate may be further coated with an additional layer (e.g., a film layer) that may be made from, for example, PVA, acetic acid, glycerin, lecithin, chitosan, etc., or any combination thereof. Each possibility is a separate embodiment. Additionally or alternatively, in some embodiments, the substrate may be attached / bonded to an additional layer that may include, for example, cotton fibers (pretreated with a cationic agent and NaOH).

[0071] In some embodiments, the substrate may further comprise or be coated with amino acids, proteins, and / or lipids. In some embodiments, the permeable substrate may comprise wool, cotton, silk, paper, and / or mixed fabrics. In some embodiments, the permeable substrate may be made from wool, cotton, silk, cellulose (e.g., in the form of filter paper), and / or mixed fabrics. In some embodiments, the substrate may be coated with a coating mixture. In some exemplary embodiments, a cotton substrate may be coated with a coating mixture comprising PVA and chitosan. In some exemplary embodiments, a cellulose coffee filter substrate may be coated with a betaine solution (e.g., 8%).

[0072] As used herein, "wool" is a textile fiber obtained from animals, primarily sheep. Wool contains primarily proteins, such as keratin, and may also contain lipids, which distinguishes it from other natural fibers, such as cotton. Wool has the ability to absorb liquids.

[0073] In some embodiments, the substrate may include synthetic textile fibers, such as, for example, plastic fibers, nylon fibers, polypropylene fibers, polypropylene nonwovens, etc. In some embodiments, the synthetic fibers may include combinations of synthetic textile fibers and natural fibers, such as cotton, wool, silk, etc., in any suitable / desired range. In some embodiments, the synthetic fibers may include combinations of polyurethane / cotton fibers, such as, for example, polyurethane / cotton, in the range of 10% / 90% to 90%-10%. In some embodiments, the synthetic fibers may include any type of suitable material combination.

[0074] According to some embodiments, the indicator composition is associated with the permeable substrate by being attached or coupled to the permeable substrate. In some embodiments, the attachment of the indicator composition and the permeable substrate is performed to minimize / insignificant leakage of the indicator composition from the substrate (e.g., less than about 10000 ppm, less than about 100 ppm, less than about 10 ppm, less than about 1.0 ppm, less than about 10%, less than about 5%, less than about 1%, etc.). In some embodiments, the attachment of the indicator composition and the permeable substrate is performed to ensure essentially undetectable or no leakage of the indicator composition from the substrate, as further exemplified below. In some embodiments, the indicator composition is irreversibly associated with the permeable substrate. In some embodiments, the indicator composition is covalently bonded to the permeable substrate. In some embodiments, the indicator is irreversibly attached to the permeable substrate. In some embodiments, the indicator is reversibly attached to the substrate.

[0075] In some embodiments, at least one indicator composition is associated with the substrate. In some embodiments, two or more indicator compositions may be associated with the substrate. In some embodiments, when multiple (e.g., at least two) indicator compositions are associated with the substrate, each indicator composition may be similar, identical, or different in terms of distribution, indicator type, composition, and / or concentration. In some embodiments, when multiple indicator compositions are associated with the substrate, each indicator composition may be associated with a distinct spatial region of the substrate. In some embodiments, the multiple indicator compositions may be substantially uniformly or evenly distributed on the substrate. In some exemplary embodiments, when multiple indicator compositions (e.g., two indicator compositions) are used, the indicators in each composition may each be of a different type (i.e., each capable of identifying or recognizing a different pathogen) and may be the same or different concentrations. In some exemplary embodiments, when multiple indicator compositions (e.g., two indicator compositions) are present, the indicators in each composition may be of the same type but different concentrations, thereby allowing a quantitative (or at least semi-quantitative) assessment or indication of the level of spoilage. In some embodiments, as described in more detail below, the indicator composition may include multiple indicators.

[0076] In some embodiments, the indicator composition includes at least one indicator and at least one carrier. In some embodiments, the carrier may be any one or more of a solvent, an acid, a base, and a diluent. In some embodiments, the carrier is an oil. In some embodiments, the carrier is or includes water or an aqueous solution. In some embodiments, the indicator composition may include multiple indicators that are the same, similar, or different in terms of type and / or concentration. In some exemplary embodiments, when multiple indicators (e.g., two indicators) are present in the indicator composition, the indicators may each be of a different type (i.e., each capable of identifying or recognizing a different pathogen) and may be of the same or different concentration. In some exemplary embodiments, when multiple indicators are present in the indicator composition (e.g., two indicators), the indicators may be of the same type but of different concentration. In some embodiments, the indicator composition may be an emulsion of the indicator and a vegetable oil and may include one or more optional adjuvants (e.g., for indicators that are not soluble in water).

[0077] As used herein, the term "about" may be used to specify a value of an amount or parameter (e.g., pore size, molecular weight) that is within a continuous range of values ​​near (and including) a given (stated) value. According to some embodiments, "about" can specify a value of a parameter to be between 80% and 120% of a given value. For example, the term "about 1" is equivalent to the term "0.8 to 1.2". According to some embodiments, "about" can specify a value of a parameter to be between 90% and 110% of a given value. According to some embodiments, "about" can specify a value of a parameter to be between 95% and 105% of a given value.

[0078] In some embodiments, the food product is in a liquid form, such as milk and non-milk / vegan milk. In some embodiments, the food product is in a semi-liquid form, such as soft cheese and gelatin. In some embodiments, the food product is in a solid form, such as chicken breast, beef, fish (including fish meat), plant-based food products, etc.

[0079] According to some embodiments, during use of the device, the indicator component within the indicator composition, or the indicator composition, may at least partially or temporarily (at any time) contact the food product to which the indicator device is attached / associated.

[0080] In some embodiments, the indicator composition may include a non-toxic, edible, food product grade indicator. In some embodiments, the indicator is selected from the group consisting of pH indicators, sulfhydryl compounds with sulfonate functionality, and nitrogen. In some embodiments, the indicator includes ninhydrin, phthalocyanine, coumarin, and / or trifluoroacetyl derivatives. In some embodiments, the indicator includes an azo dye. In some embodiments, the indicator includes indigo carmine.

[0081] In some embodiments, the at least one indicator is selected from a microbial indicator, a redox indicator, and a pH indicator. In some embodiments, the at least one indicator is a pH indicator. In some embodiments, the at least one indicator is a microbial indicator.

[0082] In some embodiments, the microbial indicator is devoid of quaternary ammonium salts and quaternary ammonium moieties.

[0083] In some embodiments, at least one microbial indicator provides a detectable indication in the presence of a microbial population above a predetermined threshold, hi some embodiments, at least one microbial indicator provides a detectable indication in the presence of a population of a particular type / species of microorganism / pathogen.

[0084] In some embodiments, the at least one microbial indicator is methyl red, methyl orange, indigo carmine, bromophenol blue, carmoisine red, tartrazine, bromocresol green, brown HT, methylene blue, alizarin, bromocresol purple, chlorophenol red, nitrazine yellow, bromoxylenol blue, neutral red, methyl orange, bromothymol blue, cresol red, phenol red, methyl red, bromocresol blue, indigo carmine, carmoisine red, tartrazine, bromocresol green, methyl orange, methylene The microbial indicator may be selected from the group consisting of erythrocyanine, citrus blue, sunset yellow FCF, citrus red 2, methylene blue, brown HT, amaranth, ponceau 4R, quinoline yellow, carmoisine, patent blue V, green S, brilliant blue, indigotine, fast green FCF, erythrocyanine, allura red AC, orange B, scarlet GN, sudan II, acid orange 20, annatto, caramel color, carmine, elderberry, lycopene, paprika, turmeric, tetrazolium salts, resazurin, ninhydrin, phthalocyanine, coumarin, trifluoroacetyl derivatives, or any combination thereof. Each possibility is a separate embodiment. In some embodiments, at least one microbial indicator comprises an azo dye.

[0085] In some embodiments, the at least one indicator is selected from bromothymol blue, cresol red, phenol red, methyl red, bromocresol blue, indigo carmine, carmoisine red, tartrazine, bromocresol green, bromophenol blue, and methyl orange.

[0086] In some embodiments, the composition further comprises at least one transition metal moiety, hi some embodiments, the transition metal moiety comprises Cr(III).

[0087] As used herein, the term "indicator" refers to any material that can change at least one characteristic (e.g., color change, spectroscopic change, colorimetric change, fluorometric change, and / or electrochemical change) with a change in a characteristic in its environment. For example, color change includes, but is not limited to, losing color (discoloration) (e.g., a colored indicator becomes white, colorless, or substantially transparent); and gaining color (e.g., a white, colorless, or substantially transparent indicator becomes colored). Environmental characteristics may include, for example, pH, the amount of oxygen in a microbial bacterial population, and / or the type of microbial bacterial population. Thus, in some exemplary embodiments, pH indicators and microbial indicators are examples of materials used as indicators in the device of the present invention. There may be an overlap between microbial indicators and pH indicators, since a change in the pH of a food product may be caused when a threshold amount of microorganisms or microbial by-products (e.g., nitrates, nitrites, sulfur, and sulfates) are produced. Exemplary indicators include colored conjugated organic molecules that change color in response to a change in pH, which results in a change in pi-conjugation of the indicator molecule; and colored conjugated organic molecules that change color in response to an enzymatic or microbial reaction, which results in a change in pi-conjugation of the indicator molecule.

[0088] It should be noted that while microorganisms may or may not be harmful, bacterial waste products may be unpleasant tasting or even harmful.

[0089] In some embodiments, the pH indicator provides a detectable mark within any pH applicable for indication of freshness or spoilage of a food product. In some embodiments, the pH indicator provides a detectable mark within a pH range of 3.0 to 9.0. In some embodiments, the pH indicator provides a detectable mark within a pH range of 4.0 to 9.0. In some embodiments, the pH indicator provides a detectable mark within a pH range of 5.0 to 9.0. In some embodiments, the pH indicator provides a detectable mark within a pH range of 8.0 to 9.0. In some embodiments, the pH indicator provides a detectable mark within a pH range of 7.5 to 8.0. In some embodiments, the pH indicator provides a detectable mark within a pH range of 7.0 to 7.5. In some embodiments, the pH indicator provides a detectable mark within a pH range of 6.3 to 7.0. In some embodiments, the pH indicator provides a detectable mark within a pH range of 5.7 to 6.3. In some embodiments, the pH indicator provides a detectable mark within a pH range of 5.0 to 5.7.

[0090] In some embodiments, the microbial indicator provides a detectable indication in the presence of an amount of bacteria above a certain threshold.

[0091] In some embodiments, the threshold as determined by the U.S. Food and Drug Administration (FDA) is about 10 5 ~10 7 (e.g., 10 million) colony forming units (CFU) / g, i.e., CFU / g, to facilitate compliance with food spoilage safety standards. Thus, food products having measured microbial levels above the FDA standard are considered unsafe for consumption and should therefore be discarded immediately. In some embodiments, any type of standard for food spoilage may be determined based on the number, concentration, amount, and / or type of microorganisms in a food sample.

[0092] In some embodiments, the microbial indicator provides a detectable indication in the presence of a compound associated with the presence of an amount of a microorganism (eg, bacteria, fungus, etc.) above a certain threshold.

[0093] In some embodiments, the microbial indicator is not an antimicrobial material. In some embodiments, the compound is the result of microbial degradation of a food product. In some embodiments, the compound is a carboxylic acid. In some embodiments, the compound includes lactic acid and / or butyric acid. In some embodiments, the compound is an amine, nitrate, nitrite, sulfur and / or sulfate. In some embodiments, the compound includes the degradation products of proteins and amino acids, such as putrescine, cadaverine, ammonia. In some embodiments, the compound is a volatile compound, including, for example, 1-octanol, 1-decanol, 1-dodecanol, 2-heptanone, 2-nonane, 2-undecanone, 3-methyl-butanoic acid, 3-methylbutanal, 3-hydroxy-2-butanone, indole. Also, in some embodiments, the microbial population can include any type of bacterial population, including, for example, aerobic, anaerobic, psychotropic and / or mesophilic microorganisms. In some embodiments, the microbial population includes pathogenic bacteria. In some embodiments, the microbial population may include any type of microorganism that may affect food spoilage, such as fungi (including yeasts and molds). For example, Paenibacillus is a spore-forming bacterium found in spoiled milk. It is responsible for spoilage of milk and also causes curdling of milk. Paenibacillus also contributes flavors in various other food products. Paenibacillus can survive the extreme conditions of pasteurization and subsequent cooling in its spore state, allowing it to survive in milk and other food products.

[0094] In some embodiments, the microbial population comprises a fungus. In some embodiments, the fungus comprises Saccharomyces cerevisiae and / or Hansenula anomala. In some embodiments, the microbial population comprises a yeast.

[0095] In some embodiments, the at least one microbial indicator is selected from the group consisting of methyl red, methyl orange, bromophenol blue, indigo carmine, carmoisine red, tartrazine, bromocresol green, and combinations thereof.

[0096] In some embodiments, at least one indicator composition comprises an active ingredient adapted to change color within a defined color range dependent on the concentration of a compound associated with food spoilage.

[0097] In some embodiments, at least one indicator is a colorimetric indicator.

[0098] As used herein, the term "colorimetric indicator" refers to, for example, a colorimetric indicator of the detection (e.g., interaction) of compounds characteristic of spoiled food products or the detection of such spoilage (e.g., pH, O 2 "color" refers to an indicator that can change color, including, for example, from a colored state to colorless (e.g., white), in response to the sensing of a condition related to a signal (such as a change in signal level).

[0099] In some embodiments, a color indicator can be provided that includes any one or more of a first color (e.g., green) indicating freshness, a second color (e.g., red) indicating spoilage, and a third color (e.g., yellow) indicating that the food product is about to spoil. The manufacturer can provide a specific date or time frame during which the food product is still fresh, but until after the specified date or by the end of the time frame, the food product will be considered to have spoiled.

[0100] In some embodiments, the detectable indication may include any one or more of the following indications: a freshness indication, a spoilage indication, and a warning indication, the latter of which may be a warning for a period of time before spoilage, e.g., a few days before the food product spoils, thereby providing a user with an indication of the time until the food product loses its freshness and / or spoils.

[0101] In some embodiments, the detectable indication is a colored detectable indication, hi some embodiments, the detectable indication comprises a color change in the visual spectrum. Table 1 provides exemplary pH transition ranges in aqueous environments for several indicators.

[0102] [Table 1]

[0103] In some embodiments, the at least one indicator is selected from the group consisting of methyl red, methyl orange, bromophenol blue, indigo carmine, carmoisine red, tartrazine, and bromocresol green. Each possibility is a separate embodiment of the present invention.

[0104] In some embodiments, the indicator device is biocompatible. In some embodiments, the indicator device is non-antimicrobial (microbe compatible). In some embodiments, at least one indicator in the indicator composition of the device is biocompatible. In some embodiments, at least one indicator is non-antimicrobial (microbe compatible). In some embodiments, at least one indicator does not include quaternary amines and quaternary amine moieties. In some embodiments, at least one indicator does not include quaternary ammonium salts and quaternary ammonium moieties. According to some embodiments, the indicator or mixture of indicators does not include quaternary amine salts as part of the composition, so they do not adhere as antimicrobials and do not inhibit / retard spoilage. In some embodiments, certain epoxy quaternary salts (e.g., 2,3-102 epoxypropyltrimethylammonium chloride, betaine, lecithin) are utilized to link the indicator (e.g., dye) to the substrate (e.g., cotton / cellulose / wool / nylon / silk / paper, etc.) backbone.

[0105] In some embodiments, the particular instruction may be specific to a particular pathogen.

[0106] In some embodiments, a food product is provided that includes an indicator device attached thereto, said indicator device being (a) Substrate and (b) an indicator composition, at least a portion of which is bound to a substrate, the substrate being configured to contact (directly or indirectly, temporarily or permanently) a food product (or at least a food sample), the indicator composition being configured to generate a detectable signal upon reaction with a portion of the food product or a component secreted therefrom, the signal being indicative of, for example, food spoilage.

[0107] In some embodiments, a method for detecting food spoilage of a food product is provided, the method being disclosed herein and comprising the steps of placing an indicator device comprising a substrate and an indicator composition attached to the permeable substrate in contact (e.g., in fluid (gas or liquid) contact) with the food product, allowing at least a portion of the food product, or a component secreted therefrom, to contact and react with the indicator composition, and upon detecting a detectable signal from the indicator device, determining that the food product is spoiled.

[0108] Reference is now made to FIG. 9, which illustrates a perspective view of an indicator device that allows for unidirectional food sample passage, according to some embodiments. Shown in FIG. 9 is an exemplary indicator device 20, further coupled / including a passageway 24, which is configured to allow for unidirectional passage of a food sample 26 to the indicator 20 of the indicator device. The passageway 24 can assume at least two positions, namely, a closed position (represented as 24) that does not allow passage from the indicator to the food product, and an open position (represented as 24') that allows passage from the food sample 26 to the indicator 22 (specifically toward the indicator). The unidirectional movement is facilitated by the passageway 24, which, as shown in FIG. 9, includes an internal bearing that allows for unidirectional fluid passage (i.e., from the food sample side toward the indicator side). The passageway may be coupled, attached, formed, or reversibly coupled to the indicator device.

[0109] In some embodiments, there is provided an indicator device configured to be attached to a food package containing a food product and to detect spoilage thereof, the indicator device comprising: (a) a substrate comprising an indicator composition; (b) at least one passageway configured to permit unidirectional passage of a portion of the food product or a component secreted therefrom to the indicator device; The indicator composition is configured to generate a detectable signal upon reaction with the portion of a food product, or the component secreted therefrom.

[0110] In some embodiments, the indicator device is impermeable. An impermeable indicator device refers to a device that does not allow the movement of an ingredient into the device unless at least one passageway is open. In some embodiments, the device does not essentially allow the movement of an ingredient or a detectable level of the ingredient from the device to the food product. In some embodiments, the indicator device is permeable.

[0111] In some embodiments, the pass is a single pass (i.e., occurs only once). In some embodiments, the pass may be repeated any number of times. In some embodiments, the pass is unidirectional.

[0112] In some embodiments, as detailed above, the indicator composition may be irreversibly, or at least more than 80% of it may be attached / bound to the substrate.

[0113] In some embodiments, at least one passageway allows unidirectional and / or single and / or multiple passes of at least a portion of the food product and / or components secreted therefrom to / into / towards the indicator device.

[0114] In some embodiments, the at least one passageway comprises at least one capillary.

[0115] In some embodiments, at least one passageway comprises a plurality of capillaries.

[0116] As used herein, the term "plurality" refers to two or more.

[0117] The use of one or more capillaries allows for spontaneous unidirectional flow of liquid from the food product (i.e., "food sample") to the indicator device, driven by capillary forces. The liquid may be a food product (or a part thereof), if the food product is a liquid such as milk, or may be a liquid secreted from a food product, such as meat (beef, chicken, fish, ham, etc.). The food sample carried to the indicator contains a microbial signature of the food product, thereby allowing its identification / determination / indication when interacting with the indicator. In some embodiments, the food sample may contain at least one reactant molecule or entity that reflects food spoilage. In some embodiments, the food sample may contain one or more reactant molecules produced in the food product that are indicative of food spoilage. In some embodiments, the food sample contains a microbial signature representative of the food product.

[0118] In some embodiments, the indicator composition is attached to a substrate and at least one passageway is a closed passageway that can be temporarily opened when desired.

[0119] As used herein, the term "temporarily open" refers to a brief, single opening to allow migration of a food portion or a component secreted therefrom (food sample) into the indicator device and then resealing at least one passageway, thereby preventing further migration into the indicator device.

[0120] The terms "brief" and "briefly" as used herein refer to a short period of time during which passage between the food product and the indicator device is permitted. Stated differently, these terms refer to a short period of time during which the passage between the food product and the indicator device is open. The short period of time may be in the range of milliseconds, seconds, minutes to 1 second. For example, the short period of time may be in the range of 1 second to 2 minutes.

[0121] In some embodiments, the closed passageway, which can be temporarily opened when needed, includes a single-use safety syringe component configured to draw a food portion or ingredient secreted from a food product into the indicator device in one go. In some embodiments, the closed passageway includes a single-use safety syringe component configured to draw a portion of a food product or an ingredient secreted from a food product into the indicator device in one go. In some embodiments, the safety syringe component includes, but is not limited to, a push-through breakable safety syringe, a rotating syringe retractable safety syringe, and a release ring breakable safety syringe.

[0122] In some embodiments, the closed passageway, which can be temporarily opened when required, comprises a displacement element configured to briefly open the closed passageway through displacement, thereby allowing transfer of a portion of the food product or a component secreted from the food product to the indicator device.

[0123] In some embodiments, the displacement is a rotation.

[0124] In some embodiments, the indicator composition is encapsulated within a substrate. In some embodiments, the substrate is an impermeable capsule. In some embodiments, the passageway is a closed passageway configured to open temporarily when required. In some embodiments, the closed passageway comprises an infiltration unit configured to briefly open the closed passageway and infiltrate the capsule, thereby allowing migration of a portion of the food product or a component secreted from the food product to the indicator device.

[0125] In some embodiments, a food package is provided that includes a food product, the package including an indicator device attached thereto, the indicator device including a substrate having an indicator composition.

[0126] In some embodiments, there is provided a food package configured to hold / contain / package a food product, the package including an indicator device attached thereto, the indicator device comprising: (a) a substrate having an indicator composition; (b) at least one passageway configured to allow unidirectional passage of a food sample (a portion of a food product, or a component secreted therefrom) into the indicator device, wherein the indicator composition is configured to generate a detectable signal upon reaction with the food sample, the detectable signal being indicative of the freshness of the food product.

[0127] The detectable signal may indicate that the food product is spoiled or not spoiled due to the presence / amount of any type of microorganism, e.g., bacteria, fungi, yeast, mold, pathogenic or non-pathogenic, depending on the type of activity of the indicator composition with the food product. Thus, the food product or indicator device may further include an index, such as a numerical value or a color scale, that indicates a correlation between the detected signal and freshness and / or spoilage.

[0128] Thus, in some embodiments, the detectable signal is indicative of a food product that has spoiled. In alternative embodiments, the detectable signal is indicative of a food product that has not spoiled. In some embodiments, the detectable signal is indicative of a food product that is fresh. In some embodiments, the detectable signal is qualitative. In some embodiments, the detectable signal is quantitative. In some embodiments, the detectable signal is qualitative and / or quantitative. In some embodiments, the detectable signal is indicative of the type and / or species and / or species of the microbial population. In some embodiments, the detectable signal may indicate whether the microorganism is a pathogen.

[0129] In some embodiments, a method for detecting food spoilage or freshness of a food product is provided, the method comprising the steps of placing / positioning an indicator device in contact with a food product, the indicator device comprising a substrate comprising at least one indicator composition and at least one passageway configured to allow a single / unidirectional passage of a food sample (a portion of the food product, or a component secreted therefrom) through the indicator device; allowing the food sample to contact the indicator composition in the indicator device via the at least one passageway; and determining whether the food product has spoiled based on detection of a detectable signal from the indicator device, the signal being indicative of food spoilage or freshness.

[0130] In some embodiments, a method is provided for detecting food spoilage or freshness of a food product, the method comprising the steps of placing / positioning an indicator device in contact with a food product, the indicator device comprising a substrate comprising an indicator composition and at least one passageway configured to allow a single or unidirectional passage of a food sample (a portion of the food product, or a component secreted therefrom, into the indicator device); allowing the food sample to contact the indicator composition in the indicator device via the at least one passageway; and determining whether the food product has spoiled upon detecting a detectable signal.

[0131] According to some embodiments, a system for detecting food spoilage of a food product is provided, the system including at least one food package configured to hold the food product and an indicator device coupled to the food package.

[0132] According to some embodiments, the food product may be selected from a liquid food product, a soft food product, a semi-rigid food product, a rigid food product, and an aqueous food product, each possibility being a separate embodiment.

[0133] According to some embodiments, the food package (also called food packaging) may be selected from any type of suitable package, including, but not limited to, for example, containers, vials, boxes, trays, cans, pallets, bags, wraps, etc. In some embodiments, the food package may be made from any suitable material or combination of materials, including, for example, plastic, cardboard, carton, metal, glass, nylon, cotton, fabric, etc., or any combination thereof. Each possibility is a separate embodiment. In some embodiments, the wrap is a nylon (saran) wrap. In some embodiments, the package may be in any condition, for example, the package may or may not contain oxygen, may or may not be evacuated, may include a controlled environment of mixed parts such as nitrogen and oxygen, or any combination thereof.

[0134] According to some embodiments, the indicator device may be integrally formed with the food package, for example, the indicator device may be incorporated into the food package during or after manufacture of the food package.

[0135] According to some embodiments, the indicator device may be permanently associated with the food package.

[0136] According to some embodiments, the indicator device may be temporarily associated with the food package.

[0137] According to some embodiments, the indicator device may be coupled to an exterior region of the food package.

[0138] In some embodiments, the indicator device may be the food package itself, for example, the indicator device may comprise a nylon substrate containing a suitable indicator, which is then used as a food package, for example by being wrapped around a food product.

[0139] Reference is now made to Figures 10A-E, which show perspective views of indicator devices and systems including the same, according to some embodiments. The devices and systems shown in Figures 10A-E show a pressure-initiated, one-way food sample passageway, where the passageway is opened by pressing the device, thereby allowing the food sample to enter the indicator. When pressure is released, the passageway is sealed / closed, and vice versa. Figure 10A shows a perspective view of an exemplary indicator device 50, including a substrate 52 and an indicator composition 54. The device may further include an outer cover, as shown in Figure 10B. Shown in Figure 10C is a system 60 including a food package 62 (shown in Figure 10C as an exemplary liquid container) and an indicator device 50 coupled thereto. In the example shown in Figure 10C, the indicator device is coupled to the front wall of the food package at the lower region / portion of the food package. Shown in Figures 10D-10E are enlarged internal views of a cross section of the device 50, with the passageway portion coupled to the inner wall 58 of the food package 62. In Figure 10D, the passageway is closed and, as shown in Figure 10E, when the indicator device is pressed at its outer region (e.g., side 57, outside the food package), the inner region of the passageway is opened, allowing the food sample 59 to pass towards the indicator device 50. Thus, when the pressure is released, the passageway may be closed / resealed, thereby allowing unidirectional passage / flow of the food sample from the food package towards the indicator device, particularly to and allowing interaction with the indicator composition, providing an indication as to the freshness / spoilage of the food product.

[0140] Reference is now made to Figures 11A-B, which show perspective views of an indicator device and a system including the same, according to some embodiments. The device and system shown in Figures 11A-B show pressure-initiated unidirectional food sample passage, where pushing the device opens a passageway, thereby allowing the food sample to enter the indicator. When pressure is released, the passageway is sealed / closed, and vice versa. Figure 11A shows a perspective view of a system 100, including a food package 104 and an indicator device 102 coupled thereto. In the example shown in Figures 11A-B, the food package is in the form of a carton, and the food product may be any suitable food product, such as a liquid food product (e.g., milk). In the example shown in Figures 11A-B, the indicator device 102 is coupled to the front wall of the food package, at a lower region / portion of the food package. Additionally, a close-up view of the device 102 coupled to the food package 104 is shown. As shown, the device includes a substrate and indicator composition 110 and a passageway 108 (allowing for unidirectional food product passage, as described in more detail below) and fits over an opening 106 in the wall of a food package 104. FIG. 11B shows a side close-up view of a cross section of the device and food package showing the interior wall 112 of the food package, with the passageway 108' having the sealed opening 106 in a closed position, and with external pressure applied to the device 102, the passageway 118" will be in an open position, allowing for the passage of food product from the interior cavity / volume of the food package to the indicator device 102, and in particular to the indicator composition. Thus, when pressure is released, the passageway may be closed / resealed, thereby allowing for unidirectional passage / flow of the food sample from the food package towards the indicator device, and in particular to the indicator composition, allowing for interaction therewith, providing an indication as to the freshness / spoilage of the food product.

[0141] Reference is now made to FIG. 12, which illustrates a perspective view of an indicator device and a system including the same, according to some embodiments. The device and system illustrated in FIG. 12 illustrates a pressure-initiated, unidirectional food sample passageway, which is opened by pressing the device, opening an internal opening in the food package, thereby allowing the food sample to enter the indicator. When pressure is released, the passageway is sealed / closed, and vice versa. FIG. 12 illustrates a perspective view of a system 150, including a food package 154 and an indicator device 152 coupled thereto. In the example illustrated in FIG. 12, the food package is in the form of a carton, and the food product may be any suitable food product, such as a liquid food product (e.g., milk). In the example illustrated in FIG. 12, the indicator device 102 is coupled to the front wall of the food package at a lower region / portion of the food package. Additionally, a close-up side view of a cross section of the device and food package is shown showing the inner wall 162 of the food package 154, whereby the indicator device passageway 156' projects into the interior volume / cavity of the food package 154 and can assume a closed (sealed) position 156' or an open position 156". External pressure applied to the device 152 facilitates a change in position of the passageway 156, in the open position allowing the passage of food product in the food package from the interior cavity / volume of the food package to the indicator device 152 (particularly its indicator composition). Thus, when pressure is released, the passageway may be closed / resealed, thereby allowing unidirectional passage / flow of the food sample from the food package towards the indicator device, particularly to the indicator composition and allowing interaction therewith, providing an indication as to the freshness / spoilage of the food product.

[0142] Reference is now made to Figures 13A-B, which show perspective views of indicator devices and systems including the same, according to some embodiments. The devices and systems shown in Figures 13A-B show unidirectional food sample passage initiated by capillary forces, which is facilitated by a capillary tube that fluidly connects the food product and the indicator device. Once the indicator is filled (or saturated) with the food sample, there is minimal or no further fluid flow between the indicator device and the food product. Figure 13A shows a perspective view of a system 200 including an indicator device 202 coupled to a food package 204. In the example shown in Figure 13A, the food package is in the form of a carton, and the food product may be any suitable food product, such as a liquid food product (e.g., milk). In the example shown in Figure 13A, the indicator device 202 is coupled to the front wall of the food package at the top region / portion of the food package. Additionally, Figure 13A shows an expanded view of the device 202, showing the substrate and indicator composition 212 and a portion of the passageway 206. 13B shows a cross-sectional interior view of a food package 204 including a capillary passage system 206 that facilitates unidirectional passage of a food sample from an interior volume 208 of the food package to an indicator device located on an exterior wall / surface of the food package. That is, the capillary passage allows for unidirectional passage / flow of the food sample from the food package towards the indicator device, particularly towards and allowing interaction with the indicator composition to provide an indication as to the freshness / spoilage of the food product.

[0143] Reference is now made to Figures 14A-B, which show perspective views of indicator devices and systems including the same, according to some embodiments. The devices and systems shown in Figures 14A-B show a one-way food sample passage facilitated by a valve unit, where one valve is open and one valve is closed, and by pivoting / rotating an actuation button, the open valve is closed and the closed valve is opened. Such a mechanism allows for one-way (one-way) stepwise passage of the food sample from the food container to the indicator device. Once the food sample has passed, the indicator device may be locked for additional passes, if desired. Figure 14A shows a cross-sectional internal view of a food package 254 of the system 250, which is coupled to an indicator device 252. In the example shown in Figure 14A, the food package is in the form of a carton, and the food product may be any suitable food product, such as a liquid food product (e.g., milk). In the example shown in Figure 14A, the indicator device 202 is coupled to the front wall of the food package at the top region / portion of the food package. Additionally, FIG. 14A shows the rear area and passageway 256 of the substrate and indicator composition 262. FIG. 14B shows the passageway valve units 258 or 260 of the passageway 256 in different configurations, specifically, an open configuration (258' / 260') or a closed configuration (258" or 260"). Changing the configuration of the valve units is facilitated, for example, by manually rotating an external button on the face of the indicator device 252. The valve unit passageway thus allows for unidirectional passage / flow of the food sample from the food package towards the indicator device (specifically the indicator composition) and allows for interaction therewith to provide an indication regarding the freshness / spoilage of the food product.

[0144] Reference is now made to Figures 15A-C, which show perspective views of indicator devices and systems including the same, according to some embodiments. The devices and systems shown in Figures 15A-C show a one-way food sample passage from a solid / semi-solid food product to the indicator device. In Figure 15A, an exemplary indicator device 300 is shown configured to be placed / positioned on a food package. The device 300 includes a substrate 302 and an indicator composition 304 bonded / coupled / attached to the substrate. Shown in Figures 15B-C is an indicator device 350 positioned / placed on a food package 310 holding a solid food product 312 (e.g., meat). The device 350 includes a substrate and an indicator composition (similar to device 300) and further includes or is coupled to a passageway 320. The passageway 320 is shown in an enlarged view, and is configured to allow movement of the food sample from the food toward the indicator composition of the indicator device. The passageway is shown in the form of a tube / channel / capillary, and the interaction between the food sample and the food product passageway is facilitated, for example, by applying pressure or by pushing the indicator device to establish contact between the passageway and the food product. Figure 15B shows the passageway before it interacts with the food product, and Figure 15C shows the passageway after it interacts with the food product. The passageway allows for unidirectional movement of the food sample from the food product to the indicator device, and in particular to the indicator composition. In some embodiments, the indicator device includes an area / chamber, and said food sample penetrates / migrates into the area / chamber via a dedicated tube / channel / capillary passageway.

[0145] According to some embodiments, the indicator device may be located / placed inside the food product and the color indication may be transferred / conveyed to the surface by optical (e.g., lens), mechanical, electronic, or chemical means.

[0146] In some embodiments, the indicator device has a top surface / face and a bottom (lower) surface / face, with the top surface configured to face the environment and the bottom surface configured to face the food product (or a food package containing the food product). In some embodiments, the characteristics of the surfaces may be different (e.g., with respect to transparency) or similar.

[0147] In some embodiments, the detectable signal produced by the indicator composition is viewable through a top surface of the indicator device.

[0148] In some embodiments, the indicator device is in the form of a flat card.In some embodiments, the indicator device is in the form of a sticker.In some embodiments, the indicator device is in the form of a patch.

[0149] As used herein, the term "sticker" refers to a flat, relatively thin device, preferably having an upper side and a lower side, the lower side being configured to face the food product and the upper side being configured to face the environment. In some embodiments, the lower side of the sticker, or a portion thereof, is adhesive or includes an adhesive area that keeps the sticker underneath in contact with the food product. In some embodiments, prior to use, one or more adhesive surfaces are irreversibly covered with a suitable film.

[0150] In some embodiments, the food spoilage is microbial spoilage.

[0151] In some embodiments, the food sample (i.e., a portion of the food product, or a component secreted therefrom) comprises reactant molecules or entities found or produced in the food product as the food product undergoes spoilage. In some embodiments, the reactant molecules or entities are or represent the microbial content / microbial population / microbial signature in the food sample (and thus in the food product). In some embodiments, the microbial population (microbial signature) comprises the type, species, amount, concentration, etc. of microbial entities in the food product. In some embodiments, the microbial signature may change over time and may be indicative of food spoilage.

[0152] In some embodiments, the detectable signal indicates that the food product is not spoiled. As used herein, "not spoiled" may refer to a fresh food product. In some embodiments, the absence of a detectable signal indicates that the food product is spoiled.

[0153] In some embodiments, the detectable signal indicates that the food product has spoiled. In some embodiments, the absence of a detectable signal indicates that the food product has not spoiled.

[0154] In the description and claims of this application, the words "include" and "have" and forms thereof are not limited to the configuration within a list with which the words may be associated.

[0155] Those skilled in the art will readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The examples provided herein are representative of preferred embodiments, are illustrative, and are not intended to limit the scope of the invention. EXAMPLES

[0156] [Example 1: Detection of food spoilage using indicators contained in wool (concentration 50 ppm)]

[0157] As detailed below, a permeable substrate made from wool fabric was dyed with indigo carmine by a simple one-step process in a slightly acidic aqueous solution.

[0158] A piece of wool cloth (approx. 65cm) 2 and / or about 8 g each) were immersed in indigo carmine dye solutions having dye concentrations ranging from 50 to 250 ppm and further containing citric acid (1%). The solution / wool ratio (w / w) was 4:1 to 5:1. Dyeing was carried out at 85-90°C for 6-7 hours. At the end of the dyeing process, the wool pieces were thoroughly rinsed with water and dried at room temperature.

[0159] Wool pieces dyed with indigo carmine dye solution containing 50 ppm dye were immersed in whole milk (about 3.5% lipid) contained in vials (one dyed wool piece per vial) and the vials were kept sealed (Figure 1A). The samples were incubated at room temperature (25-27°C) for t=0, X: about 30 hours, and Y: about 48 hours (Figures 1A-C, respectively).

[0160] As can be seen, the intense blue color of indigo carmine in the fresh milk sample shown in Figure 1A gradually faded (Figure 1B) and finally after 24-36 h, the milk sample spoiled and thus the intense blue color disappeared as shown in Figure 1C.

[0161] Example 2: Detection of milk spoilage using indicators contained in wool.

[0162] Figures 2A and 2B show two fresh milk samples, each containing a piece of wool stained with indigo carmine solution at a concentration of 60 ppm, before and after 36-40 hours of incubation at room temperature (25-27 °C). In contrast to the blue color of the indicator in the fresh milk sample (Figure 2A), the milk in Figure 2B is already spoiled, as reflected by the beige / brown color of the indicator.

[0163] Example 3: Detection of food spoilage in chicken and red meat samples using indicators contained in cotton. Fresh chicken breast and portions of lean meat were wrapped in cotton cloth stained with indigo carmine (50-60 ppm in the staining solution) that had been pretreated with a neutral cationic solution containing 2,3-epoxypropyltrimethylammonium chloride as the cationic agent, or with a basic cationic solution additionally containing approximately 0.036 M KOH (Figures 3A and 3B, respectively), left panel = chicken; right panel = beef.

[0164] As shown in Figures 3C and 3D (corresponding to Figures 3A and 3B, respectively), after 30-36 h of spoilage of red meat and chicken at room temperature, the intense color of the indicator bound to the cotton fabric decayed.

[0165] Example 4: Detection of food spoilage in dairy products using indicators contained in cotton.

[0166] 4A and 4B show fresh samples of cottage cheese wrapped in cotton pretreated with a neutral cationic solution and then stained with indigo carmine dye at t=0 and 28 hours, respectively, after incubation at room temperature.

[0167] As shown in FIG. 4B, the intense blue color of the indicator disappeared completely in the spoiled cottage cheese samples.

[0168] Example 5: Detection of soy milk spoilage using indicators contained in cotton

[0169] Figures 5A-5C show cotton fabrics stained with indigo carmine dye after pretreatment with a neutral cationic solution, shown in the state after immersion in fresh soy milk (Figure 5A, t = 0) and after incubation at room temperature for X: approximately 28-30 h and Y: approximately 48 h (Figures 5B and 5C, respectively).

[0170] As shown in Figure 5B, the intense blue color of the indicator faded to a beige / brown color in moderately spoiled soy milk after 28-30 h of incubation at room temperature (Figure 5B) and disappeared completely in the spoiled soy milk sample after 48 h of incubation at room temperature (Figure 5C).

[0171] Example 6: Detection of food spoilage in dairy and meat products using indicators contained in cotton.

[0172] The indicator device (freshness sticker (FS)) was combined with 5 ml of cottage cheese and 5 ml of chicken breast (mince).Furthermore, the indicator device (in the form of a patch / sticker) was attached to a packaged piece of fresh fish meat (salmon) and incubated at room temperature (25°C) until a colour change was observed.

[0173] The pH and CFU / ml were measured at the start (t0) and at the time of discolouration (TP).

[0174] The results are shown in Table 2 below, Figures 6A-D and Figures 7A-B. [Table 2]

[0175] The results show that all indicator devices changed colour from blue to white within one day of the experiment in the food products, namely cottage cheese (Figures 6A-B), chicken breast (Figures 6C-D) and fish (Figures 7A-B).

[0176] 7A-B, a packaged fish chunk 2 is shown having an indicator device 4 attached thereto. Also shown is an indicator 6 (which binds to a substrate) that changes color from a dark color in a fresh sample (FIG. 7A) to a lighter color (6') in a spoiled sample (FIG. 7B). The results demonstrate the ability of the indicator device to provide a visual indication as to the freshness / spoilage of a food product.

[0177] Thus, the results show that the indicator device can indeed be used for reliable detection of food spoilage, and that the indicator is inert and therefore does not affect the occurrence of food spoilage.

[0178] Example 7 Preparation of indicator devices with additional coatings or intermediary layers

[0179] Preparation of the intermediary layer: 100% cotton fabric was treated in an aqueous solution consisting of tap water, cationic agent (2,3-102 epoxypropylethylammonium chloride) and sodium hydroxide, incubated at 80-120°C for 30-180 minutes, and then dried.

[0180] Substrate and indicator preparation: 100% cotton fabric was treated with an aqueous solution consisting of tap water, cationic agent (2,3-102 epoxypropylethylammonium chloride) and sodium hydroxide, incubated at 80-120°C for 30-180 minutes, and then dried. The treated cotton was then dyed with an indicator composition containing indicators in a concentration range of 10-100 ppm in an aqueous solution. The indicator composition contains one or more types of indicators, for example, a mixture of 10-100 ppm indigo carmine, 10-50 ppm indigo carmine and 5-50 ppm tartrazine. Dyeing was carried out at 85-100°C.

[0181] Device Fabrication: If an intermediary layer is used, following drying of the intermediary layer and substrate, the substrate and intermediary layer are adhered together using an approved food contact glue based on a PVA solution and additives such as propylene glycol, lecithin, glycerol, and / or chitosan.

[0182] If a coating layer is used: After the substrate is dried, it is coated with a film layer containing a mixture of PVA, acetic acid and propylene glycol, lecithin, glycerol, and / or chitosan.

[0183] Device performance testing (indicator leakage): All experiments were performed on food products purchased from various market chains. At the start of the experiment, the expiration date was recorded and the food product was sampled for general bacterial counts. The indicator device was attached to a slice of food product (e.g., chicken breast) and immediately evacuated when indicated. The evacuated food product with the device was kept at refrigerator temperature and monitored daily for decolorization.

[0184] On the day of bleaching, samples were taken for total bacterial counts and visual indicator breakthrough on the food products was recorded.

[0185] Total bacterial count protocols were applied according to established Israeli and US Food and Drug Administration standards.

[0186] Example 8. Detection of microbial growth on fresh or autoclaved food samples using the indicator device.

[0187] Several experiments were conducted to demonstrate the relationship between the visual indication of the indicator composition of the indicator device and the presence of microorganisms in a food product.

[0188] Chicken breast samples were used as the food product. An indicator device containing an indicator composition of 20 ppm indigo carmine was placed on a slice of fresh or sterilized (autoclaved) chicken breast. The samples were kept at refrigerator temperature (4°C) and monitored for color change (indicating food spoilage) and total bacterial counts at the time of indicator device bleaching (1-20 days after the start of the test).

[0189] The results are shown in Figures 16A-B. The device (402) on fresh chicken fillets (404) changed color after the incubation period, as did the spoiled (CFU = Log 5.1 / gr (Figure 16A)) chicken fillets 404'. In contrast, for the same incubation period, the device (412) on sterile slices 414, 414' (Figure 16B) did not change color and no bacterial growth was detected. Note that autoclaving killed all bacteria on the food products 414, 414' (Figure 16B).

[0190] The results show that, indeed, the colour indication (in this example, a colour change) of the indicator composition of the indicator device occurs due to the presence and development of microorganisms on the food product.

[0191] Example 9: Determining the location of indicator devices on met food products

[0192] To determine if a representative area existed in the chicken breast food product for placement of the indicator device, the levels of bacteria in different areas were examined (rear area vs. anterior area vs. apical area). Total bacterial counts were measured on the day of purchase (0) and day 2, as well as several days after the start of the experiment. All samples were kept at refrigerator temperature along the entire experiment. The results shown in Figures 17A-B indicate that no significant differences were identified in the levels of bacteria at the beginning and throughout the experiment, regardless of whether the anterior, apical, or posterior areas were sampled.

[0193] Next, the difference in bacterial levels on the outside of the slices versus the inside of the slices was tested. The results are shown in Figure 17C-D. In chicken fillets held at refrigerator temperature for several days, the bacterial levels on the outside ("top" part) were found to be significantly higher than the bacterial levels on the inside ("inside" part). Outside part CFU=Log7.7 / gr vs. inside part CFU=Log6.4 / gr.

[0194] Taken together, these experiments indicate that the most representative area for placement of the indicator device is the outer portion of the slice from the front, as this area represents the greatest amount of bacteria to which the indicator composition of the device can react, and is also the most visible area from the consumer's point of view.

[0195] Example 10: Detection of food spoilage using indicator devices at various spoilage levels

[0196] Indicator devices containing various indicator compositions were tested for their ability to provide an indication of food spoilage at various levels of spoilage (ie, various levels of microorganisms) in various forms of food packaging.

[0197] I: Indicator composition containing 10 ppm indigo carmine To determine the CFU at the time of color change, the indicator devices were placed on a simple food package (polyethylene wrap) and placed with a vacuum pack to determine the CFU at the time of color change.

[0198] The device was calibrated to decolorize at spoilage levels for use with vacuum packaging. In this example, the device was prepared as described in Example 7 (including the media layer and excluding the top cover). The results are shown in Figures 18A-C. As shown, the indicator device changes color at defined spoilage levels while using vacuum packaging (Figures 18B, 18C). Discoloration occurs at high values ​​of CFU and the meat actually spoils when using a simple polyethylene wrap (Figure 18A, showing results from two experiments (Exp. 1, Exp. 2)).

[0199] Additionally, to improve uniformity of color change time points, a top cover can be used to reduce the coefficient of variation (CV), for example, from CV=13% to CV=7.7% for an average count of Log CFU=5.4 / gr. The type of top cover can reduce the CV (variability between test samples). Samples that showed lower variation (CV=7.7%) were approved food contact top covers.

[0200] The results show that discolouration does indeed occur at defined spoilage levels with a relatively high degree of uniformity.

[0201] II: Indicator composition containing 10 ppm indigo carmine and 5 ppm tetrazine

[0202] The device was prepared as described above. The experiment was carried out at refrigerator temperature and all breast slices were packaged under vacuum. The results are shown in Figure 18D. The results show that the color change point occurs at a Log CFU value = 5.4 (CV = 8%). These results are consistent with the former, showing high uniformity and reproducibility.

[0203] III: Indicator composition containing 15 ppm indigo carmine

[0204] In this example, a device was prepared as described in Example 7 (including a coating layer). The device was tested with chicken fillets using vacuum packaging. The experiment was carried out at refrigerator temperature. The experiment found that the time of discoloration occurred an average of 1.25 days after the expiration date of the food product. The results were similar to those obtained for the indicator device including the mediating layer (as in Experiments I and II). Furthermore, the results showed that no leakage was observed for any of the indicator devices.

[0205] Taken together, the results of this study demonstrate that the color change occurs with high uniformity and reproducibility at defined spoilage levels without detectable levels of indicator leakage into the food sample, further demonstrating and embodying the advantageous properties of the indicator device in providing a safe, reliable, and accurate indication of food spoilage.

[0206] While the present invention has been disclosed with reference to specific embodiments, it will be apparent that other embodiments and modifications of the present invention may be devised by those skilled in the art without departing from the true spirit and scope of the invention, and it is intended that the appended claims be construed to include all such embodiments and equivalent variations.

[0207] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure pertains. In case of conflict, the patent specification, including definitions, shall prevail. As used herein, the indefinite articles "a" and "an" mean "at least one" or "one or more," unless the context clearly dictates otherwise.

Claims

1. 1. An indicator device for detecting food spoilage in a food product, comprising: a substrate, and b. an indicator composition comprising at least one indicator; The indicator device, wherein the indicator composition is bound to the substrate, the substrate is configured to contact a food product, and the indicator composition is configured to generate a detectable signal upon reaction with at least a portion of the food product or a component secreted therefrom.

2. 10. The indicator device of claim 1, wherein the substrate comprises textile fibers, plastics, nylon, woven fibers, woven fabrics, nonwoven fabrics, artificial polymers, blended fibers, biodegradable polymers, cotton, wool, linen, oligosaccharides, polysaccharides, gels, modified starches, metals, aluminum, paper, carton, wood, silk, plant materials, clay, adhesives, ointments, bucks, oils, leather, carbon and graphite felt, carbon fibers, or any combination thereof.

3. The indicator device of claim 1 , wherein the substrate further comprises one or more additional layers.

4. 4. The indicator device of claim 3, wherein the additional layer comprises or is made from textile fibers, plastic, nylon, woven fibers, nonwoven fibers, cotton, wool, silk, cellulose starch chitosan-PVA, cellulose-PVA crosslinked film, or any combination thereof.

5. The indicator device of claim 3 , wherein the additional layer is permanently or temporarily bonded to the substrate.

6. The indicator device of claim 3 , wherein the additional layer is sewn, glued, and / or welded to the substrate.

7. 10. The indicator device of claim 1, wherein the spoilage is microbial-associated spoilage.

8. 10. The indicator device of claim 1, wherein the at least one indicator is selected from a microbial indicator, an azo dye, a food dye, a natural food dye, a thiazine dye, a redox indicator, a pH indicator, an oxygen indicator, a solvatochromic dye, or any combination thereof.

9. 10. The indicator device of claim 1, wherein the at least one indicator is provided in a concentration such that it changes at least one detectable property in response to spoilage of the food sample, the property comprising a spectroscopic, colorimetric, fluorescent and / or electrochemical property.

10. 10. The indicator device of claim 1, wherein the at least one indicator is provided in a concentration such that it changes color in response to spoilage of the food sample.

11. 10. The indicator device of claim 1, wherein the at least one indicator is a bacterial indicator configured to provide a detectable indication of the presence of a bacterial population above a predetermined threshold, the presence of a secreted volatile compound above a threshold, the presence of a particular type of bacterial population, or any combination thereof.

12. The indicator device of claim 1 , wherein the indicator composition comprises a plurality of indicators.

13. 10. The indicator device of claim 1, wherein the substrate has associated therewith a plurality of indicator compositions.

14. 10. The indicator device of claim 1, wherein the detectable signal is qualitative or quantitative.

15. 10. The indicator device of claim 1, wherein the indicator device is or is included in a food product package configured to at least partially hold or encase the food product.

16. An indicator device as described in claim 1, wherein the indicator device is configured to be in direct contact with a food product.

17. 1. A method for detecting food spoilage in a food product, comprising: placing a product indicator device according to any one of claims 1 to 16 in contact with the food product; contacting at least the food sample and / or secreted substances of said food product with said indicator; determining whether the food product has spoiled upon detecting a detectable signal produced by the indicator if or when the food sample has spoiled.

18. 1. A system for detecting food spoilage in a food product, comprising: At least one food package configured to hold or package a food product; and A system comprising an indicator device according to any one of claims 1 to 16 coupled to the food package.

19. 20. The system of claim 18, wherein the food product is selected from a pourable food product, a liquid food product, a soft food product, a semi-hard food product, a solid food product, a hard food product, and an aqueous food product.

20. 20. The system of claim 18, wherein the indicator device is integrally formed with or permanently or temporarily associated with the food package.

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