Systems and methods for preservation of anaerobic microorganisms

Abstract

The invention relates to methods for preserving the viability of anaerobic microorganisms in foodstuffs such as milk comprising storing the foodstuffs in a sealable storage container comprising an oxygen scavenging system. The invention also relates to an oxygen scavenging system comprising a body (522) comprising a first end, a second end (526) and a first interior compartment, a lid adapted to close the first end of the body, a gas permeable, liquid impermeable membrane (527), said membrane being suitable for closing the second end of the body, and an oxygen scavenging material (OSM), said OSM being suitable for placement in the first interior compartment.

Description

[0001] SYSTEMS AND METHODS FOR PRESERVATION OF ANAEROBIC MICROORGANISMS

[0002] FIELD OF THE INVENTION

[0003] The invention relates to an oxygen scavenging system and methods for preserving the viability of an anaerobic microorganisms, particularly in foodstuffs.

[0004] BACKGROUND OF THE INVENTION

[0005] Human breast milk is the optimal nutrition for infants. In addition to essential nutrients (including proteins, fat, carbohydrates, vitamins and minerals) and bioactive components, breast milk also contains a diverse array of bacteria, fungi, and other microorganisms, which make up the milk microbiome. The anaerobic microorganisms present in human breast milk refers to the community of microorganisms that thrive in the absence of oxygen. The largest contribution of the maternal microbiome to that of her infant is from the microbes within her breast milk, contributing to over 30% of the first infant microbiome (Bogart et al, 2023, Cell and Host Microbe Volume 31 , Issue 3, 8 March 2023, Pages 447-460. Over 200 bacterial species, both aerobic and anaerobic, are known to be present in human breast milk belonging to various taxonomic groups, including Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria. Some commonly identified genera include Bifidobacterium, Lactobacillus, Clostridium, Staphylococcus, Streptococcus, Veillonella, and Prevotella.

[0006] While aerobic (oxygen-requiring) bacteria are also present, anaerobic microorganisms play a significant role in shaping the composition and function of the breast milk microbiome. These diverse microbes, which colonise the infant's gut, are critical for developing the immune system of the infant and offsetting disease. Some facultative or strict anaerobic species belonging to Lactobacillaceae and Bifdobacteriaceae have been associated with and exploited for their health benefits in the prevention and treatment of many pathological conditions and to improve overall health. These anaerobic microorganisms contribute to the establishment and maintenance of a healthy infant gut microbiota. They help in the digestion and metabolism of complex carbohydrates present in breast milk, produce short-chain fatty acids (SCFAs) as metabolic byproducts, and support the development of the growing infant's immune system. Hence, these microorganisms are typically referred to as probiotics, which are live microorganisms that, when administered in adequate amounts, confer a health benefit on the host.

[0007] Anaerobic bacteria in breast milk have also been found to produce enzymes that break down human milk oligosaccharides (HMOs), which are complex sugars abundant in breast milk but indigestible by the infant. By metabolising these HMOs, the anaerobic microorganisms provide a nutritional source for themselves and produces metabolites that can help modulate the developing immune system and promote the growth of other beneficial bacteria, such as Bifidobacteria, Lactobacillus and Limosilactobacillus, in the infant's gut. Additionally, the anaerobic microorganisms in breast milk may contribute to immune modulation and protection against pathogens. Certain strains of bacteria additionally produce antimicrobial compounds, such as bacteriocins, that inhibit the growth of harmful bacteria. Furthermore, the presence of specific anaerobic bacteria in breast milk has been associated with a reduced risk of allergic diseases and improved gut health in infants.

[0008] Other components of breast milk which support the gut microbiome health and infant nutrition include antioxidants such as vitamin C. Antioxidants are vital for protecting molecules involved in metabolism from losing electrons and therefore functionality. They work by giving up some of their own electrons to reactive oxygen species (ROS), neutralising ROS damaging effects. When breast milk is stored in ambient oxygen these antioxidants immediately start reacting with ROS and so the antioxidant capacity of breast milk reduces. As the antioxidant capacity decreases, the environment becomes less favourable for survival of probiotic bacteria, reducing their availability to support the infant's metabolism after milk consumption.

[0009] The composition of the anaerobic microorganisms in breast milk can vary among individuals and may be influenced by factors such as maternal health, diet, and environmental exposures. Hence, it will be appreciated that there are many complex interactions between breast milk microbiota, maternal factors, and the health outcomes of infants.

[0010] Literature suggests the mode of breastfeeding - whether mothers expressed milk with or without a pump - is a key determinant of the milk microbiota composition. More specifically, a study by Moossavi et al. (Cell and Host Microbe Volume 25, Issue 2, 13 February 2019, Pages 324-335) shows that compared to direct feeding of breast milk, pumping resulted in lower microbiota richness and is associated with the accumulation of potential pathogens and the depletion of Bifidobacterium. Other studies have shown that the levels of important Bifidobacterium that co-occur in mothers’ milk and their infants’ stool are reduced when infants receive pumped breast milk. It is not clear why pumped breast milk contains a different microbiome. One possibility is the lack of retrograde inoculation whereby the infant's oral microbiome is transferred to breast milk. Another possibility is that some species of bacteria are sensitive to oxygen and become depleted in the process of pumping and subsequent storage.

[0011] While oxygen scavenging systems are known in the art, there is no disclosure of an oxygen scavenging system for preserving the anaerobic microorganisms naturally present in milk within a home setting. US 2014 / 0312000 discusses an oxygen absorbing cap for preserving liquids such as wine. JP H04367472 discusses an oxygen scavenger attached to the mouth of a container. US 4287995 A discusses a container sealing member for a container used for preserving foodstuffs such as soy sauce, Japanese sake, sauce, wine, beer, juice, vinegar, etc. WO 2014 / 041391 discusses a canister for containing an active material, such as a desiccant. US 5092914 A discusses an oxygen-absorbing cartridge that floats in a liquid. Schwab et al (Front. Microbiol., 20 Nov 2019) discusses the use of an anaerobic jar for cultivating the microbiota of human breast milk within a laboratory setting. The protocol discussed in Schwab et al necessitates the use of an Anaerocult A system placed within an anaerobic chamber.

[0012] Hence, it will be appreciated that there is a need to improve the preservation of the anaerobic microorganisms within breast milk that has been freshly expressed. In particular, due to the variation in the makeup of the microbiome between individuals and the profound effect that this has upon infant health, there is a need to provide improved devices and methods for preserving expressed breast milk at the point of use.

[0013] SUMMARY OF THE INVENTION

[0014] The present inventors have developed novel systems and methods for preserving the viability of beneficial anaerobic microorganisms in a medium. In addition, the systems also show utility in preserving the antioxidant content of foods and beverages beyond the usual expected shelf life.

[0015] In a first aspect, the invention provides a method for preserving the viability of anaerobic microorganisms comprised within milk, said method comprising storing the milk in a sealable storage container wherein the sealable storage container comprises an oxygen scavenging system (OSS).

[0016] The sealable storage container is selected from: a vessel, a bottle, a sealable bag or a pouch. Suitably, the sealable storage container is an infant feeding bottle.

[0017] The milk comprises mammalian milk. Suitably, the milk is human breast milk. More suitably, the breast milk is freshly expressed.

[0018] The anaerobic microorganisms comprise species selected from anaerobic bacteria, optionally selected from Bifidobacteriaceae and / or Lactobacillaceae, suitably wherein the Bifidobacteriaceae are selected from Bifidobacterium infantis, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium animali, Bifidobacterium dentium, s and / or Bifidobacterium bifidum.

[0019] In a second aspect, the invention provides an oxygen scavenging system (OSS) for use with a sealable storage container to preserve the viability of anaerobic microorganisms in a medium comprised within, said OSS comprising: a) a body comprising a first end and a second end, wherein the first and second ends are open; and wherein the body defines a first interior compartment; b) a lid adapted to seal the first end of the body; c) a gas permeable, liquid impermeable membrane, said membrane being suitable for closing the second end of the body; and d) an oxygen scavenging material (OSM), said OSM being suitable for placement in the first interior compartment.

[0020] In an embodiment, the OSS further comprises a gasket, said gasket suitable for providing a seal between an opening of the storage container and a cap suitable for closing the opening of the storage container. In an embodiment, the gasket surrounds the body and extends radially outwardly therefrom. In another embodiment, the gasket comprises a resilient material, optionally wherein the resilient material comprises a silicone rubber. In another embodiment, the gasket extends radially outwardly from the body by a distance of at least 1 cm, optionally at least 2cm, suitably at least 3 cm. In an embodiment, the gas permeable, liquid impermeable membrane is comprised of a hydrophobic material, or comprises a hydrophobic coating applied thereto. In another embodiment, the gas permeable, liquid impermeable membrane further comprises a hydrophobic coating.

[0021] In an embodiment, the oxygen scavenging material (OSM) is selected from one or more of: ferrous iron powder with sodium chloride; optionally with activated carbon; a ferrous carbonate in conjunction with a metal halide catalyst; ferrous iron oxide; unsaturated fatty acid; sodium hydrogen carbonate; citric acid and ascorbic acid.

[0022] In an embodiment, the amount of OSM for placement in the first interior compartment is at least 1 g, at least 2 g, suitably at least 5 g, optionally at least 10 g; and at most 30 g, suitably at most 40 g; optionally at most 50 g.

[0023] In an embodiment, the OSS further comprises a sleeve. In an embodiment, the sleeve is frustoconical.

[0024] In an embodiment, the OSM is placed in the first interior compartment, the first end is sealed by the lid and the second end is sealed by the gas permeable liquid impermeable membrane. In an embodiment, the second end is sealed by two gas permeable liquid impermeable membranes separated by a space between them.

[0025] In an embodiment, the gas permeable liquid impermeable membrane is covered by a gas impermeable, liquid impermeable covering. In an embodiment, the gas impermeable, liquid impermeable covering can be peeled off by a user.

[0026] In an embodiment, the storage container is selected from: a vessel, a bottle, a sealable bag or a pouch. In a preferred embodiment, the storage container is an infant feeding bottle.

[0027] In an embodiment, anaerobic microorganisms comprise species selected from anaerobic bacteria, optionally selected from Bifidobacteriaceae and / or Lactobacillaceae, suitably wherein the Bifidobacteriaceae are selected from Bifidobacterium infantis, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium animalis, Bifidobacterium lactis, Bifidobacterium dentium and / or Bifidobacterium bifidum.

[0028] In an embodiment, the OSS further comprises an oxygen indicator. In an embodiment, the oxygen indicator is a dye.

[0029] In an embodiment, the OSS selectively promotes the growth of probiotic anaerobic microorganisms and suppresses the growth of pathogenic microorganisms in a medium, optionally wherein the medium is mammalian milk, optionally human breast milk.

[0030] In an embodiment, the OSS preserves the antioxidant content of a medium, such as milk.

[0031] In a third aspect, the invention provides a method for preserving the shelf life of freshly expressed milk, said method comprising storing milk in a storage container wherein the storage container comprises an oxygen scavenging system (OSS). In an embodiment, the OSS is of the second aspect. In a fourth aspect, the invention provides an infant feeding bottle comprising an oxygen scavenging system (OSS). In an embodiment, the OSS is of the second aspect of the invention.

[0032] In a fifth embodiment, the invention provides a sealable container for preserving the viability of anaerobic microorganisms comprised within, the container comprising the OSS of the second aspect of the invention.

[0033] In a sixth embodiment, the invention provides a cap for sealing a sealable container for preserving the viability of anaerobic microorganisms comprised within. The cap is adapted to receive the OSS of the second aspect of the invention.

[0034] In a seventh aspect, the invention provides a method for selectively promoting the growth of probiotic anaerobic microorganisms and suppressing the growth of pathogenic microorganisms in freshly expressed human breast milk, said method comprising storing the breast milk in an infant feeding bottle comprising an OSS of the second aspect of the invention.

[0035] Within the scope of this application, it is expressly intended that the various aspects, embodiments, examples and alternatives set out in the preceding paragraphs, in the claims and / or in the following description and drawings, and in particular the individual features thereof, may be taken independently or in any combination. That is, all embodiments and / or features of any embodiment can be combined in any way and / or combination, unless such features are incompatible.

[0036] BRIEF DESCRIPTION OF THE DRAWINGS

[0037] One or more embodiments of the invention will now be described, by way of example only, with reference to the accompanying drawings, in which:

[0038] Figure 1 shows an embodiment of the oxygen scavenging system.

[0039] Figure 2 shows a storage container with a cap comprising an embodiment of the oxygen scavenging system according to the invention.

[0040] Figure 3 shows the cap of the storage container of Fig. 2

[0041] Figure 4 shows an alternative embodiment of the oxygen scavenging system placed within a sealable container.

[0042] Figure 5 shows alternative embodiments of the oxygen scavenging system.

[0043] Figure 6 shows a cap comprising an oxygen scavenging system according to the invention.

[0044] Figure 7 shows an embodiment of the oxygen scavenging system with an integrated lid.

[0045] Figure 8 shows an alternative embodiment of the oxygen scavenging system placed on a side wall of a storage container. Figure 9 shows the placement of an alternative embodiment of the oxygen scavenging system in a sealable container

[0046] Figure 10 shows the CFU count of a probiotic breast milk-isolated Bifidobacterium breve strain exposed to oxygen at room temperature over time. Mean values with standard deviation are plotted. The CFU count of the strain decreases with increasing duration of exposure to oxygen..

[0047] Figure 11 shows the OD readings over time of a probiotic oxygen-sensitive Bifidobacterium infantis strain grown in MRS broth at 37 °C, both with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Growth is enhanced when stored in the device compared to when not stored in the device.

[0048] Figure 12 shows the OD readings over time of a second probiotic oxygen-sensitive Bifidobacterium infantis strain grown in MRS broth at 37 °C, with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Growth is enhanced when stored in the device compared to when not stored in the device.

[0049] Figure 13 shows the OD readings over time of a probiotic oxygen-tolerant Limosilactobacillus fermentum strain grown in MRS at 37 °C, with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Growth is not affected when stored in the device compared to when not stored in the device.

[0050] Figure 14 shows the OD readings over time of opportunistic bacterium Staphylococcus aureus grown in MRS at 37 °C, both with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Growth is inhibited when stored in the device compared to when not stored in the device.

[0051] Figure 15 shows the CFU counts of a probiotic oxygen-sensitive Bifidobacterium breve strain inoculated into pooled pasteurised breast milk and stored at 4°C over time, both with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Strain viability is enhanced when stored in the device compared to when not stored in the device.

[0052] Figure 16 shows the CFU counts of a probiotic oxygen-sensitive Bifidobacterium longum strain inoculated into pooled pasteurised breast milk and stored at 4°C over time, both with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Strain viability is enhanced when stored in the device compared to when not stored in the device.

[0053] Figure 17 shows the CFU counts of a probiotic oxygen-sensitive Bifidobacterium bifidum strain inoculated into pooled pasteurised breast milk and stored at 4°C over time, both with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Strain viability is preserved when stored in the device compared to when not stored in the device.

[0054] Figure 18 shows the CFU counts of a probiotic oxygen-tolerant Limosilactobacillus fermentum inoculated into pooled pasteurised breast milk and stored at 4°C over time, both with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Strain viability is preserved when stored in the device compared to when not stored in the device.

[0055] Figure 19 shows the CFU counts of a probiotic oxygen-sensitive Bifidobacterium infantis (strain 1) inoculated into pooled pasteurised breast milk and stored at 4°C overtime, both with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Strain viability is higher when stored in the device compared to when not stored in the device.

[0056] Figure 20 shows the CFU counts of oxygen-sensitive Bifidobacterium infantis strain spiked intro fresh pooled breast milk and stored at -20°C over time, both with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Strain viability is preserved when stored in the device compared to when not stored in the device.

[0057] Figure 21 shows bar graphs illustrating the normalised abundance of (A) Bifidobacterium bifidum and (B) Bifidobacterium animalis species in breast milk at baseline (Oh) and after 72 hours of storage at 4°C, with and without the device. Mean values with standard error are plotted. Statistical significance was assessed using a two-way t-test, with p-values indicated by asterisks (**p < 0.01). The abundance of B. bifidum and B. animalis was significantly lower when the milk was not stored with the device, whereas no significant change was observed when stored with the device, indicating preservation.

[0058] Figure 22 shows the CFU counts of pathogenic Klebsiella pneumoniae strain inoculated into pasteurised pooled breast milk and stored at room temperature over time, both with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Strain viability is reduced when stored in the device compared to when not stored in the device.

[0059] Figure 23 shows the CFU counts of pathogenic Staphylococcus aureus strain inoculated into pasteurised pooled breast milk and stored at room temperature over time, both with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Strain viability is reduced when stored in the device compared to when not stored in the device.

[0060] Figure 24 shows the antioxidant concentration in breast milk samples stored at 4°C over three days (72 hours), both with and without the device. Mean values with standard deviation are plotted. Statistical significance was assessed using a 2-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001). Antioxidant concentration is preserved when stored in the device compared to when not stored in the device.

[0061] Figure 25 shows a graph of breast milk pH with and without device during fridge incubation (25A) and at room temperature (25B).

[0062] Figure 26 shows a pie chart illustrating the participants' identification of spoilage in breast milk samples via olfactory testing. Of the 65 tests conducted, 71 % (n=46) of participants identified the milk stored without the device as showing greater signs of spoilage, while 29% (n=19) selected the milk stored with the device. This demonstrates the effectiveness of the device in reducing perceived spoilage during storage.

[0063] Figure 27 shows a graph depicting the change in the RED component of the RGB of an oxygen sensitive sticker (Emco) when placed in (A) 5cc and (B) 25cc oxygen environments in the presence of the claimed device. The oxygen scavenging ability of the device is maintained for at least ten days compared to no device.

[0064] Figure 28A shows photographs of methylene blue oxygen indicator solution filled to 15% capacity in three conditions: In vacuum (Tommee Tippee Colicsoothe Milk Air Remover) Sterifeed bottle with the device, and Sterifeed bottle without the device, captured at 0, 30, and 60-minute intervals. Fig. 28B shows the corresponding plot of pixel intensity, processed in Fiji (ImageJ). The device successfully decolourised the methylene blue solution from blue to clear within 60 minutes (indicating removal of oxygen), with most oxygen scavenging occurring by 30 minutes. Minimal changes were observed in the no-device and vacuum conditions.

[0065] Figure 29 shows photographs of methylene blue oxygen indicator solution filled in Sterifeed bottle with the device filled with different scavengers (IMPAK, MIDKU IT, OXOID) at full cartridge (FC) and half cartridge (HC) capacities at 0, 30, 60 and 90 minutes. All scavengers passed by 90 minutes, apart from the IMPAK HC condition. Photographs show flexibility and effectiveness of the cartridge design for oxygen scavenging across various absorbers. ‘Passed’ indicates the decolourisation of methylene blue from blue to clear.

[0066] Figure 30 shows photographs of methylene blue oxygen indicator solution filled to 15% capacity in bottles of varying size fitted with the device. Photographs were captured at 0-min and 60- min. The device successfully decolourised the methylene blue solution from blue to clear within 60 minutes for all bottles.

[0067] Figure 31 shows photographs of methylene blue indicator solution filled in Medela bottles fitted with the device of varying Tyvek membrane surface areas: small (S=1 .95 cm diameter), medium (M=2.2 cm diameter), and large (L=2.35 cm diameter) cartridges. The trend indicates that larger surface areas result in faster oxygen removal, with complete decolourisation (blue to clear) observed in all conditions by 60 minutes. Figure 32 shows another embodiment of the oxygen scavenging system comprising a cover to attach the oxygen scavenging system to the opening of a storage container.

[0068] DETAILED DESCRIPTION OF THE INVENTION

[0069] The systems and devices described herein are particularly useful in preservation of key components, such as anaerobic microorganisms as well as the antioxidant content of live foodstuffs, beverages and biological samples including, but not limited to, human breast milk.

[0070] Figure 1 shows an oxygen scavenging system (OSS) 100 suitable for use with a storage container to preserve the viability of anaerobic microorganisms in a medium comprised within the storage container. The OSS comprises a casing; a gas permeable, liquid impermeable membrane; an oxygen scavenging material (OSM); and an oxygen indicator.

[0071] The casing 104 comprises a first end 106 and a second end 108; and a first wall 110 and a second wall 114, both walls enclosing a first interior space 112 extending between the first and second ends 106, 108. In an alternative embodiment (not shown), the casing 104 further comprises a second interior space defined between the first and second walls 110, 114. In this embodiment, the second interior space extends at least to the second end 108 of the casing, optionally from the first end 106 to the second end 108 of the casing.

[0072] In the embodiment of Fig. 1 , the first and second ends 106, 108 of the casing 104 are open. The second end 108 of the casing 104 is adapted to accommodate the gas permeable, liquid impermeable membrane 118 such that when the membrane of a suitable size is introduced into the casing, the membrane can close the second end of the casing. In the embodiment shown in Figure 1 , the second end of the casing comprises a lip 120 extending into the first interior space 112 such that the opening at the second end is smaller than the opening at the first end of the casing. This allows the gas permeable liquid impermeable membrane to be placed securely inside the first interior space adjacent to the second end of the casing, thereby closing the second end. ‘Close’ as used herein refers to a closure of the opening with a material such that the material selectively limits the entry or exit of gases and liquids over time. The membrane of the invention selectively allows the passage of gas such as oxygen while substantially limiting the passage of liquids.

[0073] The first interior space of the casing is suitable for accommodating a body 122. The body comprises a first end 124 and a second end 126 wherein the first and second ends are open. The body further defines a first interior compartment 128.

[0074] The first interior compartment 128 extends from the first end of the body to the second end of the body. In an alternative embodiment, the first interior compartment 128 extends from a point between the first and second ends 124, 126 to the second end 126 of the body 122.

[0075] In the embodiment shown in Fig. 1 , the body further comprises a second interior compartment 130. The second interior compartment extends at least to the second end of the body, optionally from the first end to the second end of the body. The second interior compartment is located adjacent to the first interior compartment. In an alternative embodiment, the second interior compartment at least partially surrounds the first interior compartment. Optionally, the second interior compartment fully surrounds the first interior compartment.

[0076] The lid 132 is adapted to seal the first end of the casing and / or the body. ‘Seal’ as used herein refers to a closure of an opening with a material such that the material substantially limits the entry or exit of gases and liquids overtime.

[0077] In an embodiment, a sealing ring 134 is used to seal the first end of the body and / or the casing. Examples of sealing rings are known in the art including mechanical gaskets for e.g., an O-ring.

[0078] The oxygen scavenging material (OSM) is placed in the first interior compartment 128.

[0079] In the embodiment shown in Fig. 1 , an oxygen indicator can be placed in the second interior compartment 130. In an alternative embodiment, the oxygen indicator can comprise all or part of a printed image. Such an image may be affixed to a transparent portion of the oxygen scavenging system so that it is visible from the outside of the system or container.

[0080] In an alternative embodiment of the OSS, the body is an integral part of the casing.

[0081] An alternative embodiment is shown in Figure 5. In this embodiment, a body 522 comprises a first end 524 and a second end 526. The body 522 defines a first interior compartment 528. The first interior compartment 528 extends from the first end of the body to the second end of the body. The second end 526 is sealed with a gas-permeable, liquid impermeable membrane 527. In embodiments, a gas- impermeable, liquid impermeable seal 529 is applied to the gas-permeable, liquid impermeable membrane 527. The oxygen scavenging material 521 is placed in the first interior compartment 528. The first end 524 is closed with a lid (not shown). Any suitable means may be used to close the first end. In embodiments, the lid is heat sealed to the first end 524.

[0082] In embodiments, the oxygen scavenging system further comprises a gasket 525. In embodiments, the gasket is suitable for providing a seal between an opening of the storage container and a cap suitable for closing the opening of the storage container.

[0083] The gasket 525 is substantially circular, typically in the form of an annular ring. The gasket extends outwardly and enables the OSS to form a complete closure of the opening of the storage container.

[0084] In embodiments, the gasket is elastically extensible and / or deformable.

[0085] In embodiments, the gasket comprises a resilient material. Suitably, the gasket is made from a silicone or any other suitable elastomeric material, e.g., polyurethane rubber, thermoplastic polyurethane rubber, polytetrafluoroethylene, latex, polyisoprene, EPDM, SBR, etc.

[0086] In embodiments, the gasket surrounds the body 522. In such embodiments, the gasket comprises a central opening into which the OSS may be placed. In use, the gasket forms a liquid- and gas-tight seal around the body. In embodiments, the gasket is a skirt gasket. The portion of the gasket adjacent to the central opening form a skirt around the OSS when the OSS is placed within the central opening of the gasket.

[0087] In embodiments, the body 522 comprises a lip region 523 at the first end such that the diameter of the body 522 in the lip region 523 is greater than the diameter of the central opening of the gasket. This allows the gasket to hold the OSS in place. Alternatively, the lip region is present in the lid of the body 522. In another embodiment, both the first end of the body and the lid comprise the lip region when the lid is placed so as to seal the first end of the body.

[0088] In embodiments, the gasket extends radially outwardly from the body, so as to form a flange, by a distance of at least 0.5cm, optionally at least 1 cm, at least 1 .5 cm, at least 2cm, at least 2.5 cm, at least 3 cm, at least 3.5 cm, at least 4 cm, at least 4.5 cm, at least 5 cm, at least 5.5 cm, at least 6 cm.

[0089] In embodiments, the gasket can be trimmed manually to fit the opening of the storage container, as needed. This allows the OSS to be adapted to fit any suitable storage container, such as infant feeding bottles from different manufacturers.

[0090] Fig 5B shows an embodiment of the OSS of the invention without a gasket or a sleeve. The body 522 defines an interior compartment 528 comprising the OSM 521. The second end 526 of the body 522 comprises a gas permeable liquid impermeable membrane 527.

[0091] In Fig 5C, the OSS further comprises a gas impermeable liquid impermeable membrane 529. The gas impermeable liquid impermeable membrane prevents contact of the gas permeable liquid impermeable membrane with air prior to use. The user activates the OSS by removing or peeling the gas impermeable liquid impermeable membrane 529.

[0092] In an alternative embodiment shown in Fig 7, the lid 723 is attached to the body 722. In an embodiment, the attachment is via a hinge or a latching mechanism.

[0093] Fig 9 shows another embodiment of the OSS of the invention. The body 922 is placed within a sleeve 927 which is placed in the central opening of the gasket 925. Fig 9B shows the placement of the OSS into the opening of an infant feeding bottle 910.

[0094] In embodiments, the lid is an integral part of the body.

[0095] In an embodiment shown in Fig. 32, the OSS comprises a cover. The cover 3220 is adapted to receive the body 3222 and to attach to the opening of the storage container 3210. The cover is made from silicone rubber. Any suitable elastomeric material may be used for the cover (e.g., polyurethane rubber, thermoplastic polyurethane rubber, polytetrafluoroethylene, latex, polyisoprene, EPDM, SBR, etc.) which allows the coverto accommodate the body in opposition to the opening of the container. To put the OSS into use, the body is placed inside the cover (after removal of the gas impermeable liquid impermeable membrane / coating, if present), the cover is then inverted overthe opening of a storage container and attached to the opening. In embodiments, the OSS comprises a separate sleeve. The sleeve has a first end and a second end, both of which are open. The sleeve is shaped to receive the casing. In an embodiment, the sleeve is frustoconical.

[0096] The sleeve fully or partly covers the casing. In embodiments, the sleeve has a diameter smaller than the lid of the casing but larger than the body of the casing.

[0097] In embodiments, the sleeve has a lip such that the diameter of the lip is largerthan the diameter of the opening of the gasket.

[0098] In embodiments, the gasket may be stretched over the opening of the storage container. In such embodiments, the gasket is secured over the opening of the container by the elastic nature of the material. Optionally, an O-ring may be used to secure the gasket in place. In embodiments, the OSS may be placed on a side wall of the container. An example is shown in Fig. 8A. The container 800 comprises an integrated oxygen scavenging system 805 in a side wall, the oxygen scavenging system comprising an oxygen permeable membrane 801 , an oxygen scavenging material and a body 822 enclosing the oxygen scavenging material 802. The container 800 further comprises an air-tight lid 803. In an embodiment shown in Fig. 8B, the container 800 further comprises an additional layer 804 enclosing the oxygen scavenging system 805.

[0099] In another aspect, the invention provides a cap for sealing a sealable container for preserving the viability of anaerobic microorganisms comprised in a medium stored in the container. In the embodiment shown in Fig. 2, the cap 240 comprises a hole 242 suitable for receiving the oxygen scavenging system 204 of the second aspect of the invention. The OSS engages with the cap 240 with a suitable mechanism, such as a push-fit mechanism. In Fig. 2, an O-ring 234 secures the lid 232 to the first end of the oxygen scavenging system 204. The cap 240 in Fig. 2 is shown in greater detail in Fig. 3. In Fig. 3, the cap 340 comprises a hole 342.

[0100] In an alternative embodiment, the oxygen scavenging system is an integral part of the cap. As shown in Fig. 4, the cap 440 comprises a first end 406 and a second end 408. A first interior space 428 is defined between the first and second ends 406, 408. The first interior space 428 is suitable for accommodating the oxygen scavenging material 444. The oxygen scavenging material 444 may be placed directly inside the first interior space 428. In another embodiment, the oxygen scavenging material may be placed inside a film, pouch or another covering which is then placed inside the first interior space 428. The gas permeable, liquid impermeable membrane 418 is located at the second end 408 of the cap 440. The first end 406 of the cap 440 is adapted to be sealed with a lid 432. The second end 408 of the cap 440 is adapted to seal a storage container 402 for preserving the viability of anaerobic microorganisms or other components in a medium stored within the storage container. Sealing engagement of the lid 432 with the first end 406 may be through a push fit seal, or via a threaded engagement as depicted in Fig. 4.

[0101] The oxygen indicator used in the OSS is able to indicate the presence of oxygen (oxidised form or positive result) as a colour change, which is distinguishable from the condition indicating a lack of oxygen (reduced form or negative result). Examples of dyes suitable for use as oxygen indicators include resazurin, indigo carmine, methylene blue, diphenylamine, phloxine, or a combination thereof.

[0102] Gas permeable membrane refers to a membrane that is selectively permeable to gases while being impermeable to non-gaseous material. The term “permeable” or “gas permeable” means a material that allows gases, in particular oxygen (O2), to be transferred from one side of the material to the other, in either or both directions. As used herein, the related terms “breathable” and “semipermeable” are synonymous with “permeable" and the two terms can be used interchangeably herein. Typically, the material is comprised within a planar layer, sheet, film or membrane. The permeable or gas permeable material may be microporous or may be comprised within a composite material, such as a composite membrane. As such, the permeable or gas permeable material may be comprised within a layer (e.g., a selective layer), coating or surface treatment applied to a support material or gutter layer within the composite. Thin film composites of this type are thereby provided with a selective layer contributing to gas permeation which is, thus, disposed on a support layer - e.g., comprising a plurality of hollow fibres - that provides mechanical strength to the material as a whole. The permeation is directly related to the concentration gradient of the permeant (such as gas), a material’s intrinsic permeability, and the diffusivity of the permeant species in the membrane material. Examples of gas permeable membranes suitable for use with the systems and methods of the present invention include organic polymers including low-density polyethylene, high-density polyethylene for e.g., Tyvek®, silanized alumina, siloxane-based polymers, including polydimethylsiloxane (PDMS), and the like.

[0103] In an embodiment, the gas permeable liquid impermeable membrane comprises a hydrophobic coating. The term “hydrophobic material” refers to a material that repels or is resistant to water. Exemplary hydrophobic materials include polyvinylidene fluoride (PVDF), octadecyltrochlorosilane (OTS), hydrophobic polymers including, PLGA, etc.

[0104] The casing and the body may be comprised of any suitable material including lightweight plastics such as high-density polythene, polypropylene, polycarbonate or polyethylsulphone, Tritan; or polyamide, borosilicate glass, tin plate, foil or ceramics.

[0105] In use, the oxygen scavenging system is incorporated into the storage container such that the gas permeable, liquid impermeable membrane is exposed to the interior of the storage container. The interior of the container will typically comprise a portion of atmosphere that may comprise oxygen. The oxygen gas will permeate the oxygen scavenging system where is absorbed or otherwise sequestered by the oxygen scavenging material. Hence, the oxygen scavenging system is able to deplete the concentration of oxygen within the storage container and thereby render the environment within the container more suitable for the viability of anaerobic organisms that may be comprised within the medium. Depending on the OSM selected, the OSS is able to deplete the concentration of oxygen within the storage container even after short periods of exposure to atmospheric conditions by opening and closing the bottle.

[0106] In specific embodiments of the invention the oxygen scavenging system may be used to precondition the storage container prior to introduction of a medium containing one or more anaerobic organisms. Introduction of the medium may, thus, include a flush with a non-oxygenated gas at the time the medium is introduced (e.g., nitrogen flush) to ensure oxygen is not re-introduced to the pre-conditioned storage container.

[0107] In an embodiment, the OSS is produced and packaged under sterile conditions to prevent microbial contamination. Alternatively, or additionally, the OSS can be subjected to sterilisation, for e.g., by exposure to heat or chemicals. In embodiments, the OSS is encased in a covering, such as a film, to maintain its sterility.

[0108] The covering may also be used to protect the oxygen scavenger from exposure to air. It may be removed just prior to use to minimise exposure to oxygen before use.

[0109] In an embodiment, the sealable container is comprised of a material suitable for cryopreservation. ‘Cryopreservation’ refers to a process including at least one step of lowering the temperature of a biological material from a temperature that is above the freezing temperature of the biological material (or of a mixture of the biological material and a preservation composition) to a temperature that is below that freezing temperature. Cryopreservation encompasses freezing, vitrification and lyophilization. Materials suitable for manufacture of the sealable container and / or the housing of the system 200 may include stainless steel; aluminium; glass; lightweight plastics such as high-density polythene, polypropylene, polycarbonate or polyethylsulphone; or polyamide.

[0110] In an embodiment, the sealable container is a bottle. Suitably, the bottle is an infant feeding bottle. Infant feeding bottles are known in the art. An infant feeding bottle may have a two-part design and include a bottle body and a cap. The substantially hollow bottle body is configured to contain a liquid infant food, such as milk, suitably breast milk, and may include an outer screw thread on the outer wall of its upper portion that is engageable by an inner screw thread provided on an inner wall of the cap such that the cap is attachable to the upper portion of the bottle body. The cap may be a teat to facilitate feeding of the infant. The term ‘infant feeding bottle’ is interchangeable with ‘baby bottle’ and ‘nursing bottle’. Typically, infant feeding bottles can accommodate up to 330ml of liquid.

[0111] In an alternative embodiment of the invention, the sealable container may be in the form of a pouch, a bag, for example a milk collection and / or storage bag or a stool collection bag for FMT, as a lid which is compatible with specific breast milk bottles of which the cartridge can pop in / out of, as a bung to fill hungate / conical flasks where the cartridge can be fitting inside the bung to allow culture of anaerobic bacteria. In the case where the sealable container is in the form of a pouch or bag it is predominantly defined by non-rigid or semi rigid materials. Such materials may include low density polyethene, polypropylenes and various plastic films. However, the sealing member may still comprise rigid materials, such as a cap or lid into which the oxygen scavenging system may be incorporated or otherwise affixed.

[0112] In an embodiment, the oxygen scavenging system may comprise an engagement mechanism to engage the oxygen scavenging system with a container. The engagement member may be in the form of at least one circumferential rib which engages with or couples with a corresponding part on the container. Alternatively, the engagement mechanism may be a groove, a clip, a hook, a notch to allow coupling of the oxygen scavenging system to the container. In an embodiment, the engagement mechanism may be in the form of an adhesive.

[0113] In embodiments, the OSS comprises an engagement mechanism to engage the OSS with a bottle lid. Any suitable engagement mechanism may be used, for example a prong and slot mechanism seen in Fig 6A. The prong on the body 622 engages with a slot 623 in a lid 625.

[0114] In an alternative embodiment shown in Fig. 6B, the lid comprises fastening slots 627 which engages the body 629.

[0115] In some embodiments, the oxygen scavenging system is disposable. In other embodiments, the oxygen scavenging system is reusable. In reusable embodiments, the oxygen scavenging system can be sterilised or otherwise cleaned. The oxygen scavenging material can be replenished with new material, and the spent oxygen scavenging material may be reduced back to its active state for incorporation into new cartridges or sent for recycling. It is desirable to promote a sustainable circular economy that facilitates the re-use and recycling of materials utilised in the systems and methods of the invention.

[0116] In some embodiments, the amount of OSM for placement in the first interior compartment is at least 1 g, at least 2 g, suitably at least 5 g, at least 6 g, at least 7 g, at least 8 g, at least 9 g, at least 10 g; at least 12 g, at least 15 g, at least 20 g, and at most 30 g, suitably at most 40 g; optionally at most 50 g.

[0117] In use, the systems, devices and methods of the invention are suitable for preserving or improving the viability of anaerobic microorganisms. In an embodiment, the anaerobic microorganisms are anaerobic bacteria. Exemplary anaerobic bacteria include Bifidobacteria, Bacteroides and Faecalibacterium species.

[0118] Bifidobacterium, Bacteroides and Faecalibacterium genera are obligate anaerobes that contribute to a number of human health benefits. These genera are known to support digestive health by promoting the breakdown and absorption of nutrients, improving gut motility, and preventing gastrointestinal issues such as diarrhoea and constipation. They also help maintain a balanced gut microbiota, which is essential for overall digestive function. In particular, probiotic Bifidobacterium strains have been shown to modulate the immune system, stimulating the production of antibodies, enhancing immune response, and reducing the risk of certain infections. They can help regulate the immune system's inflammatory response and have been shown to be beneficial in managing conditions such as allergies and inflammatory bowel diseases. Antibiotics can disrupt the natural balance of bacteria in the gut, leading to diarrhoea. Bifidobacterium probiotics have been used to prevent or alleviate antibiotic-associated diarrhoea by restoring the microbial balance in the intestines. In respect of infant health, Bifidobacterium species are important for the establishment of a healthy gut microbiota in infants. They are commonly added to infant formulas or administered as supplements to promote the colonization of beneficial bacteria in the infant's gut and potentially reduce the risk of conditions like colic, eczema, and allergies. Nevertheless, the health benefits of Bifidobacterium species can vary depending on the specific strain, dosage, and individual factors. The system, devices and methods described herein allow for the removal of oxygen, for example, periodic or continuously, from the atmospheric environment surrounding the medium. Thus, the system, devices and methods described herein allow for the preservation of compositions and components susceptible to degradation by oxygen.

[0119] Suitably, the medium comprises a consumable foodstuff including milk, a pharmaceutical or nutraceutical composition, or a biological sample that includes a live microbial component. A live microbial component refers to live microorganisms, such as bacteria, yeast, fungi or a combination thereof.

[0120] In a specific embodiment of the invention, the medium comprises human breast milk, typically freshly expressed human breast milk.

[0121] In other embodiments the foodstuff may comprise live fermented foodstuffs - e.g. a food product that has or is undergoing fermentation by the action of live microorganisms, such as bacteria, yeast, or a combination of both. By way of example, such foodstuffs may include kombucha or kimchi, or similar fermented products. The live microorganisms present in these types of foods can have probiotic properties, supporting the balance of beneficial bacteria in the gut and promoting digestive health. Pharmaceutical or nutraceutical compositions that include a live microbial component may include probiotic supplements intended for treatment of conditions that include gastrointestinal infections or inflammatory disorders.

[0122] In other embodiments, the medium comprises antioxidants including vitamin E, vitamin C, vitamin A, polyphenols, metals such as copper, zinc and selenium, and the like. Other examples of antioxidants include coenzyme Q10, idebenone, lycopene, green tea polyphenols, siribine, resveratrol, grape seed extract, olegon grape root (Mahonia aquifolium) extract, pomegranate extract, genistein, pycnogenol, Curcumin, Curcuminoid, Tocopherol, Dunariella salina extract, or a combination thereof. The systems, devices and methods described herein are suitable for preserving the antioxidant content of media such as foodstuffs, nutraceuticals, pharmaceuticals. The systems, devices and methods described herein are particularly suitable for preserving the antioxidant content of milk, particularly expressed breast milk.

[0123] In one embodiment of the invention the total antioxidant capacity (TAC) of a portion of human breast milk is extended during storage when using the systems, containers or devices of the invention compared to when they are not used. The TAC describes the ability of antioxidants in different foods or biological derived products to scavenge harmful free radicals, such as reactive oxygen species that impair shelf-life, reduce nutritional potential and promote spoilage. Antioxidants can neutralize these free radicals and prevent oxidation. The TAC can be measured by various methods that use different chemical reactions to generate free radicals and monitor their reduction by antioxidants, including TRAP (total radical trapping parameter) assay (Young IS, (2001) J. Clin. Path 54: 339). TRAP uses 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) to generate peroxyl radicals, which are then trapped by antioxidants present in the sample being tested. Oxidation reactions are monitored by measuring oxygen consumption using an oxygen electrode and the lag time before oxygen is consumed is proportional to the TAC of the sample being tested. Hence, TAC can provide an overall assessment of the antioxidant status of food samples, but it does not reflect the individual contribution of the different antioxidants within the sample or their interactions. In specific embodiments of the invention the TAC of the sample, including food or beverage, is extended by at least 6 hours, at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 1 week, at least two weeks, at least one month, at least two months, optionally at least six months, when compared to storage in a container that does not include the claimed systems or devices.

[0124] Biological sample refers to any sample that is taken from a subject (e.g., a human being) and includes for e.g., faecal matter. In an embodiment, the system, devices and methods are used to preserve the viability of microorganisms in a faecal matter sample.

[0125] The method according to the first aspect of the invention preserves the viability of anaerobic microorganisms comprised within milk, said method comprising storing the milk in a sealable storage container wherein the sealable storage container comprises an oxygen scavenging system (OSS). In embodiments, the OSS is according to the second aspect of the invention.

[0126] The methods disclosed herein also selectively promote the growth of probiotic anaerobic microorganisms and suppress the growth of pathogenic microorganisms, in freshly expressed human breast milk.

[0127] The methods disclosed herein also serve to improve the viability of probiotic anaerobic microorganisms.

[0128] DEFINITIONS

[0129] As used herein, the term ‘comprising’ means any of the recited elements are necessarily included and other elements may optionally be included as well. ‘Consisting essentially of’ means any recited elements are necessarily included, elements that would materially affect the basic and novel characteristics of the listed elements are excluded, and other elements may optionally be included. ‘Consisting of’ means that all elements other than those listed are excluded. Embodiments defined by each of these terms are within the scope of this invention.

[0130] The term “hydrophobic material” refers to a material that repels or is resistant to water.

[0131] The terms "premature" and "low birth weight (LBW)" newborns are used interchangeably and refer to newborn babies born at less than 37 weeks gestational age and / or with birth weights less than 2500g.

[0132] The term ‘oxygen scavenging material’ as used herein refers to any material capable of gathering or removing oxygen from its environment. Suitable oxygen scavengers are e.g. oxidizable organic compound and a transition metal catalyst; ethylenically unsaturated hydrocarbon and a transition metal catalyst; a reduced form of a quinone, a photo-reducible dye, or a carbonyl compound which has absorbance in the UV spectrum; a polymer having a polymeric backbone, cyclic olefinic pendent group, and linking group linking the olefinic pendent group to the polymeric backbone; a copolymer of ethylene and a strained, cyclic alkylene; ethylene / vinyl aralkyl copolymer; ascorbate; isoascorbate; sulfite; ascorbate and a transition metal catalyst, the catalyst comprising a simple metal or salt, or a compound, complex or chelate of the transition metal; a transition metal complex or chelate of a polycarboxylic acid, salicylic acid, or polyamine; tannin; or reduced metal such as iron. Other examples include ferrous iron powder with sodium chloride, optionally with activated carbon; a ferrous carbonate in conjunction with a metal halide catalyst; ferrous iron oxide; unsaturated fatty acid; sodium hydrogen carbonate; citric acid and ascorbic acid.

[0133] An ‘oxygen indicator’ is a composition that indicates the presence or absence of oxygen. Oxygen indicators include dyes that change colour depending on the presence or absence of oxygen. Exemplary dyes include resazurin, indigo carmine, methylene blue, diphenylamine, phloxine, or a combination thereof.

[0134] “Milk” refers to milk obtained from a mammal, formula milk, or a combination of both. Mammalian milk may refer to for example bovine milk, goat milk or human milk. Formula milk, interchangeably used with infant formula, refers to manufactured milk intended as a substitute to breast milk for feeding infants. Formula milk may be derived from for example cow milk or plants such as soya and oat.

[0135] The term "anaerobic microorganism" as used herein, refers to microorganisms that can live and grow in an atmosphere having less free oxygen than tropospheric air (i.e., less than about 10%, by mol., of free oxygen). Anaerobic microorganisms include organisms that can function in atmospheres where the free oxygen concentration is less than about 10% by mol., or less than about 5% by mol., or less than about 2% by mol., or less than about 0.5% by mol.

[0136] ‘Bifidobacterium' and its synonyms refer to a genus of anaerobic bacteria having beneficial properties for humans. Bifidobacterium is one of the major taxonomic groups of bacteria that make up the gut flora of infants. Bifidobacteria are among the beneficial commensal bacteria that reside in the gastrointestinal tract and have health benefits for their hosts. Different species of Bifidobacteria include Bifidobacterium longum subspecies infantis, Bifidobacterium longum, Bifidobacterium animalis subspecies lactis, Bifidobacterium breve and Bifidobacterium bifidum. Each of these species can break down certain varieties of human milk oligosaccharides from breast milk and have been shown to be effective probiotic strains for preventing the risk of developing diseases such as necrotising enterocolitis in preterm infants and more broadly for improving gut health of full-term infants.

[0137] 'Lactobacillaceae' is a family that includes various genera such as Lactobacillus, Limosilactobacillus, Lacticaseibacillus, and Lactiplantibacillus, along with other related genera. These genera are part of a diverse group of lactic acid bacteria known for their role in fermentation and probiotic functions.

[0138] The term “pathogen” is defined to include microorganisms, bacteria, viruses, and fungi, including but not limited to psychrotrophic bacteria; lipolytic psych rotrophic bacteria; proteolytic psychrotrophic bacteria; mesophylic bacteria; Bacillus species, including B. cereus; Clostridium species, including C. perfringens and C. botulinum; Cryptosporidium species; Campylobacteria species, including C. jejuni; Listeria species, including L. monocytogenes; Escherichia species, including E. coli and pathogenic E. coli strains; Mycobacterium species, including M. paratuberculosis; Pseudomonas species, including P. fluorescens; Helicobacteria species; Yersinia species, including Y. entercolitica; Arcobacter species; Aeromonas species; Toxoplasma species, including T. gondii; Streptococcus species; Staphylococcus species, including S. aureus; Shigella species; Salmonella species, including S. enteritidis, S. Montevideo, S. typhimuriunr, Cyeiospora species, including C. cayetanensis; Cignatera species; Vibrio species; Plesiomonas species; Entamoeba species, including E. histolytica; Candida species, including C. albicans; Hepatitis viruses; Astroviruses; Calciviruses; enteric Adenoviruses; Parvoviruses; and Rotaviruses.

[0139] All references cited herein are incorporated by reference in their entirety. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0140] The invention is further illustrated by the following non-limiting examples.

[0141] EXAMPLES

[0142] Example 1 : Probiotic breast milk-isolated Bifidobacterium breve strain is sensitive to oxygen

[0143] Methods: A culture of Bifidobacterium breve isolated from breast milk was plated on BSM agar in triplicate at various dilutions. The plates were exposed to aerobic conditions for staggered time points, allowing for oxygen exposure durations ranging from 10 minutes to 4 hours. Following this, the plates were incubated anaerobically for 48 hours at 37 °C, and colony counts were quantified. The average of three replicates (n=3) is plotted for each time point, with error bars representing standard deviation. Note that some error bars are too small to be shown.

[0144] Results: The CFU count of the Bifidobacterium breve strain decreased with increasing time exposure to oxygen, indicating a decline in the strain's viability under aerobic conditions (Fig 10).

[0145] Example 2: Growth of breast milk-isolated bacteria using the device under optimal conditions

[0146] The device can enhance the growth of probiotic oxygen-sensitive and oxygen-tolerant breast milk- isolated strains, but inhibits the growth of pathogenic bacteria, under optimal growing conditions

[0147] Methods: Bacterial strains isolated from breast milk, including Bifidobacterium infantis (strain 1), Bifidobacterium infantis (strain 2), Bifidobacterium animalis, Limosilactobacillus fermentum, and Staphylococcus aureus, were cultured in 5 mL of MRS broth within 50 ml_ Falcon tubes under two conditions: with and without the device. Cultures were prepared in triplicate (n=3) for each condition and incubated at 37 °C. Optical density (OD) at 600 nm was measured over time by taking 100 pL aliquots from each Falcon tube.

[0148] Growth curves from both conditions were compared, focusing on the time to reach the stationary phase and maximum OD. The mean of the replicates is plotted for each time point, with error bars representing standard deviation. Note that some error bars are too small to be shown. Statistical significance was assessed using a two-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001).

[0149] Results: The device significantly enhanced the growth of the oxygen-sensitive Bifidobacterium infantis strains (both strain 1 and strain 2) under optimal growing conditions. Notably, these strains did not grow without the device (Fig. 11 and Fig. 12). Limosilactobacillus fermentum, known to be oxygen-tolerant, showed no significant difference in growth with or without the device, indicating that the device does not affect aerotolerant strains (Fig 13). Conversely, Staphylococcus aureus, an opportunistic pathogen, demonstrated inhibited growth in the presence of the device (Fig 14).

[0150] Example 3: Preservation of probiotic bacteria in breast milk using the device

[0151] A: The device can preserve the viability of probiotic oxygen-sensitive breast milk-isolated strains inoculated in pooled pasteurised breast milk

[0152] Methods: Bacterial strains isolated from breast milk, known for their probiotic effects, including Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium bifidum, and Limosilactobacillus fermentum, were inoculated into pooled and pasteurised breast milk and stored in a refrigerator (4 °C) for up to 6 days, with and without the device. Cultures were prepared in triplicate (n=3) for each condition. Bacteria-inoculated milk samples were plated on BSM agar at 0 hours, 48 hours, and 144 hours (6 days) and incubated anaerobically for 48 hours at 37 °C, after which colony counts were quantified. A Bifidobacterium infantis strain was also investigated using the same methods although sampled at 0 hours, 48 hours, 72 hours and 96 hours. The mean of the replicates is plotted for each time point, with error bars representing standard deviation. Note that some error bars are too small to be shown.

[0153] Statistical significance was assessed using a two-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001).

[0154] Results: All strains tested exhibited improved viability over the 6-day or 4-day storage period when stored with the device. At each time point (48 hours, and 144 hours), the CFU counts for all strains were significantly higher in samples stored with the device compared to those stored without the device. This was also shown in the Bifidobacterium infantis time points (48 hours, 72 hours, and 96 hours). The increased viability of probiotic strains in breast milk stored with the device indicates a significant preservation effect, as evidenced by the statistical analysis. This effect was demonstrated by p-values of *p* < 0.05, with several strains showing even more robust significance levels (e.g., **p < 0.01 , ***p < 0.001 , ****p < 0.0001). These results highlight the ability of the device to maintain the viability of probiotic strains in breast milk over extended refrigerated storage. These results are shown as Figs. 15 to 19.

[0155] B: The device can preserve the viability of oxygen-sensitive breast milk strains in fresh pooled breast milk stored in the freezer.

[0156] Methods: A Bifidobacterium infantis strain isolated from breast milk, known for its probiotic effects, was inoculated into pooled and pasteurised breast milk and stored in a freezer (-20 °C) for 7 days and 21 days with and without the device. The samples for each time point are stored separately to avoid the need for freeze-thawing of milk. Cultures were prepared in triplicate (n=3) for each condition. Bacteria- inoculated milk samples were plated on BSM agar at each time point and incubated anaerobically for 48 hours at 37 °C, afterwhich colony counts were quantified. The mean of the replicates is plotted for each time point, with error bars representing standard deviation. Note that some error bars are too small to be shown. Statistical significance was assessed using a two-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001).

[0157] Results: The viability of Bifidobacterium infantis declined significantly when stored in the freezer (-20°C) over time (Fig 20). A steep decline in CFU counts was observed as early as 7 days in samples without the device. However, in samples stored with the device, this reduction in viability was significantly mitigated (**p < 0.01), demonstrating the device's protective effect. By day 21 , further decline in Bifidobacterium infantis viability was observed in samples stored without the device, while CFU counts remained steady in the device-protected samples. This preservation effect of the device was highly significant, with ****p < 0.0001 compared to the no-device condition.

[0158] C: The device can preserve Bifidobacterium species present in breast milk microbiome samples

[0159] Methods: High-quality taxonomic assignments were generated from 16S amplicon sequencing of human breast milk using the CosmosID Kepler pipeline. Raw read counts assigned to species were filtered using the filterByExpr method from the edgeR package to remove species with sparse data (i.e., species with many zeros). The raw counts were normalised using counts per million (CPM) and log- transformed with a pseudo count of 0.25. To test for the preservation or depletion of bacterial abundances, normalized counts were compared using a t-test between samples collected at 0 hours and 72 hours, with and without the device, focusing on the Bifidobacterium species present in the samples.

[0160] Results: The Bifidobacterium species, Bifidobacterium animalis and Bifidobacterium bifidum, showed significant decreases in abundance (p=0.003 and p=0.005, respectively) between the 0-hourtime point and the 72-hour time point when the breast milk was not stored in the device (Fig. 21). However, these same species showed no significant reduction in abundance when the breast milk was stored in the device, indicating preservation of Bifidobacterium in the milk. Interestingly, these two species were undetectable in the 72-hour samples of milk stored without the device, suggesting that they were either completely absent or present at such low abundances that they were undetectable by sequencing, which explains the zero standard deviations in these samples.

[0161] Notably, all other Bifidobacterium species assessed, with the exception of one, showed a reduction in average abundance between the O-hour and 72-hour time points when the milk was stored without the device. However, these reductions did not reach statistical significance, likely due to the low sample size and the inherent high variation between individuals. Nevertheless, the significant results and trends observed for the Bifidobacterium species suggest a preservation of abundance in milk stored with the device and a reduction in abundance in samples not stored in the device.

[0162] Example 4: Inhibition of opportunistic pathogenic bacteria in breast milk using the device

[0163] The device inhibits viability of opportunistic pathogenic bacteria strains in pasteurised pooled breast milk.

[0164] Methods: Opportunistic pathogenic bacterial strains, including Klebsiella pneumoniae and Staphylococcus aureus were grown and inoculated into pooled and pasteurised breast milk and stored at room temperature, with and without the device. Cultures were prepared in triplicate (n=3) for each condition. Samples were collected and plated on LB agar at specific timepoints: 0, 4, 8, and 24 hours for K. pneumoniae; 0, 4, and 12 hours for S. aureus and incubated aerobically for 24 hours at 37 °C, after which colony counts were quantified. The mean of the replicates is plotted for each time point, with error bars representing standard deviation. If error bars are not visible, they are too small to be shown. Statistical significance was assessed using a two-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001).

[0165] Results: At the 4-hour time point, which aligns with the recommended guidelines for safe room temperature storage of breast milk, no statistically significant difference in bacterial viability was observed between the device and no-device conditions for Klebsiella pneumoniae (Fig 22), Staphylococcus aureus (Fig 23). This indicates that the presence of the device does not negatively affect the safety of breast milk during the early stages of storage as the device does not contribute to increased bacterial growth within the initial 4-hour period.

[0166] Beyond the 4-hour time point, however, statistically significant differences in bacterial viability were observed for all tested pathogens. The device effectively reduced the viability of Klebsiella pneumoniae and Staphylococcus aureus, resulting in significant inhibition of bacterial growth. These reductions were quantified with p-values of **p < 0.01 , ***p < 0.001 , and ****p < 0.0001 . This data demonstrates that the device substantially inhibits pathogen proliferation after 4 hours, supporting its utility in preserving the microbial safety of breast milk over extended periods of room temperature storage.

[0167] Example 5: Breast milk shelf-life increases with device

[0168] A. Breast milk antioxidant retention

[0169] Methods: Freshly expressed breast milk was collected from donors and transported to the laboratory in an icebox, with a maximum travel time of 1 .5 hours. Upon arrival, samples were collected at Oh and stored at 4°C with and without the device. Oh (n=30 donors), 72 hours with device (n=27 donors), 72 hours without device (n=26 donors). Samples from each donor were tested in triplicate for antioxidant molar concentration using the Ferric Antioxidant Status Detection Kit (Invitrogen™, ThermoFisher Scientific). Antioxidant capacity was calculated based on the standard curve provided by the kit manufacturer. Replicates that varied by more than 5% were excluded as outliers. The mean of the replicates is plotted for each time point, with error bars representing standard deviation. Statistical significance was assessed using a two-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001).

[0170] Results: Breast milk stored using the device retained significantly more antioxidant concentration after 72 hours compared to milk stored without the device at 4°C (****p < 0.0001). Additionally, there was no significant difference between the antioxidant concentration of milk stored with the device for 72 hours and that of freshly collected milk (Oh) without the device, demonstrating the device's effectiveness in preserving antioxidants over time. In contrast, there was a significant decrease in antioxidant concentration between Oh and milk not stored with the device (****p < 0.0001) (Fig. 24). B. No change in breast milk pH with device

[0171] Method: Breast milk was collected as described above in Example 2A, although this time breast milk was also stored with and without the device at room temperature (four conditions total, two with / without device stored 4°C, two with / without device stored at 22°C) Using a pH metre (HANNA HI-2210-02), breast milk pH was measured for the fridge conditions at days 0 (N=10), 1 (N=6), 2 (N=8) and 3 (N=4), and the room temperature condition on day 0 (N=10), 1 (N=10) and 2 (N=8). Using a One-way ANOVA, the mean pH value was compared.

[0172] Results: There is no significant difference in pH between conditions at each timepoint for both temperatures, with pH gradually decreasing as storage time increases. This suggests the device does not actively change the pH of breast milk compared to standard storage. While it is not significantly different, a trend could be seen where the pH of breast milk stored at room temperature decreases more rapidly than the pH in the fridge storage condition (Fig 25). This can be correlated to the growth of anaerobic bacteria, which produce lactic acid as a byproduct, resulting in a lowered pH.

[0173] C. Reduced breast milk deterioration from olfactory tests

[0174] Methods: Freshly expressed breast milk was collected from donors and transported to the laboratory in an icebox, with a maximum travel time of 1 .5 hours. The milk was then stored in the refrigerator at 4°C for seven days, both with and without the device. After the storage period, participants were given two samples of the same donors’ breast milk to smell: one sample stored with the device and one sample stored without the device. This was a single-blind experiment, where the participants were not informed which sample had been stored with the device. Participants were asked to assess the samples based solely on olfactory cues and respond to the question, "Which sample demonstrates signs of spoilage?"

[0175] In total, breast milk from 13 donors were tested, each donor had between 3-6 participants smell their milk at day 7. This totalled 65 comparisons of milk stored in either treatment arm, with 14 participants.

[0176] Results: Of the participants, 71% (n=46) identified the milk stored without the device as showing greater signs of spoilage, whereas only 29% (n=19) indicated that the milk stored with the device exhibited greater signs of spoilage. This suggests that the device effectively reduces spoilage perception over the storage period (Fig. 26).

[0177] Example 6: Oxygen Scavenger characterisation

[0178] A. Oxygen is reliably removed by the device in varying storage volumes

[0179] Method: ATCO Oxygen Indicator Stickers (supplied by Emco) were placed inside 50 mL Ffalcon tubes with 25 mL of distilled water (oxygen content: 5 cc) and empty 100 mL duran bottles (oxygen content: 25 cc) containing the device. Photographs were taken over the course of ten days to observe the colour of the sticker, which retains a bright yellow colour in the absence of oxygen and a dark blue colour in the presence of oxygen. The device was then taken out of both containers and the sticker photographed after two days. To quantify colour changes RGB values of the sticker were estimated (Imaged). Based on ATCO oxygen indicator colour scale, the red component of the RGB value decreases to 50 in the presence of oxygen and remains at 255 in the absence of oxygen. This information was used as a standard against which stickers in the experiment were evaluated.

[0180] Results: The results are shown in Fig 27. The device sustained an oxygen free environment within both the 50 mL falcon tubes and 100 mL duran bottles for 10 days, as identified by the retention of the yellow colour and RGB value of 255. In contrast, when the device was removed from both storage containers the stickers changed to the dark blue colour indicative of oxygen, with a RGB value of 50. This demonstrates the device is successfully removing all oxygen from the storage containers and that standard storage does not actively remove oxygen.

[0181] B. Methylene blue oxygen indicator confirms that the device efficiently outperforms both standard storage and vacuum storage in removing oxygen

[0182] Method: Sterifeed bottles were filled to 15% of their volume capacity with methylene blue indicator solution (475 mmol potassium hydroxide, 185 mmol glucose, 17.4 pmol methylene blue), which is blue in the presence of oxygen and clear in its absence. The 15% volume was chosen to mimic the lowest volume of breast milk that would typically be filled in the bottle, therefore reflecting the maximum milk oxygen exposure. All bottles were vortexed at maximum speed for 15 seconds and then photographed to capture the baseline blue colour produced by the oxidation reaction. The Sterifeed bottles were then sealed either with (n=3) or without (n=3) the device. For the device condition, the cartridge was filled with 9g of OS (Midukit). The bottles were then vortexed again and photographed at 30 and 60 minutes.

[0183] In parallel, a Tommee Tippee Colicsoothe Milk Air Remover (TCMAR) was filled to 15% of its capacity with the methylene blue indicator solution. The vacuum was then activated, and once complete, the TCMAR was vortexed again for 10 seconds and photographed. The solution was vortexed and photographed at 30-minute intervals. This process was repeated three times (n=3).

[0184] To quantitatively analyse the data, the photographs were processed using Fiji (Imaged). The images were first converted to 32-bit greyscale, which assigns pixel intensity values ranging from 0 (black) to 255 (white). The region of interest (ROI) for each image was selected, and the mean pixel intensity of the solution was measured to assess changes in colour intensity. These mean intensity values were then plotted over time, allowing for a quantitative comparison of the oxidation reaction across different conditions. The mean of the replicates is plotted for each time point, with error bars representing standard deviation. Statistical significance was assessed using a two-way ANOVA, with p-values indicated by asterisks (*p < 0.05, **p < 0.01 , ***p < 0.001 , ****p < 0.0001).

[0185] Results: The photographs (Fig 28A) and corresponding plot (Fig 28B) demonstrate that the device successfully decolourised the methylene blue oxygen indicator solution from blue to clear within 60 minutes, with the majority of oxygen scavenging occurring by the 30-minute mark. In contrast, minimal changes in colour intensity were observed in both the no-device and vacuum conditions, indicating little to no oxygen removal in these cases (Fig. 28).

[0186] C. Comparing device with different commercially available oxygen scavengers at varying weights Methods: Three oxygen scavengers were tested: IMPAK’s Non-Ferrous Oxygen Absorber, Midukit’s ferrous Oxygen Absorber, and Oxoid™ AnaeroGen™ absorber, with the device fitted on Sterifeed bottles. Each absorber was used to either fill a full cartridge (FC) or half a cartridge (HC) at the following weights: IMPAK, 4.6g and 2.3g, respectively; AnaeroGen, 3g and 1.5g, respectively; Midukit, 9g and 4.5g, respectively. A total of 25 mL of methylene blue indicator solution was placed in a Sterifeed bottle, and the cartridge and device were sealed. Another 25 mL of methylene blue indicator solution was placed in a standard Sterifeed bottle and sealed as a control. Each bottle was vortexed at maximum speed for 15 seconds and photographed after 10 seconds at 0, 30, 60, and 90 minutes. Each condition was repeated three times. Once the sample had passed the test, no further photos were captured for that sample at subsequent timepoints.

[0187] Results: Photographs of methylene blue indicator in different conditions show that by 30 minutes, the FC Oxoid and both the FC and HC Midukit had successfully removed all oxygen. By 60 minutes, the HC Oxoid had also removed all oxygen. At 90 minutes, the FC IMPAK scavenger had removed all oxygen, whereas the HC IMPAK did not fully remove oxygen within the timeframe. These results indicate that the Midukit was the most efficient scavenger. Overall, the findings demonstrate that the cartridge design is flexible and effective for oxygen scavenging across a range of different absorbers. The results were consistent across repeats (n=3) (Fig. 29).

[0188] D. Methylene blue oxygen indicator confirms device efficiently removes oxygen from different bottles of varying size

[0189] Methods: Five bottles of varying sizes (Sterifeed total volume = 168 mL, Avent total volume = 195 mL, Tommee Tippee total volume = 320 mL, Dr. Brown total volume = 300 mL, Medela total volume = 330 mL) were filled to 15% of their capacity with methylene blue oxygen indicator solution. Each bottle was vortexed at maximum speed for 10 seconds and photographed to record the baseline blue colour produced by the oxidation reaction. A total of 7.9 g of OS was weighed and placed into the device which was then sealed inside each bottle. After 60-minutes, each bottle was vortexed for 10 seconds and photographed. Each condition was repeated three times.

[0190] Results: The device can remove oxygen from bottles of varying sizes within 60 minutes, as indicated by the change in colour of the methylene blue oxygen indicator from blue to clear. The results were consistent across repeats (n=3) (Fig. 30).

[0191] E. Comparing device with different membrane surface area

[0192] Methods: To determine the appropriate surface area of the Tyvek membrane required for oxygen scavenging within one hour, different cartridge sizes with varying Tyvek diameters were tested using Medela bottles (total volume = 330 mL). The diameters of the Tyvek membrane in each cartridge were as follows: small cartridge (S=1 .95 cm), medium cartridge (M=2.2 cm), and large cartridge (L=2.35 cm). Medela bottles were filled to 15% capacity with methylene blue oxygen indicator solution. Each bottle was vortexed for 15 seconds and photographed at 0, 30, and 60 minutes. Each condition was repeated three times. Results: A clear trend was observed in which larger Tyvek membrane surface areas resulted in faster oxygen scavenging, as indicated by the colour change of the Methylene Blue solution from blue to clear. The large cartridge (L) showed the fastest oxygen removal, followed by the medium (M) and small (S) cartridges. Despite the differences in speed, oxygen was fully removed in all conditions by 60 minutes. The results were consistent across repeats (n=3) (Fig. 31).

[0193] Although particular embodiments of the invention have been disclosed herein in detail, this has been done by way of example and for the purposes of illustration only. The aforementioned embodiments are not intended to be limiting with respect to the scope of the appended claims, which follow. It is contemplated by the inventors that various substitutions, alterations, and modifications may be made to the invention without departing from the spirit and scope of the invention as defined by the claims.

Claims

CLAIMS1 . A method for preserving the viability of anaerobic microorganisms comprised within milk, said method comprising storing the milk in a sealable storage containerwherein the sealable storage container comprises an oxygen scavenging system (OSS).

2. The method of claim 1 , wherein the storage container is selected from: a vessel, a bottle, a sealable bag or a pouch.

3. The method of claim 1 or 2, wherein the sealable storage container is an infant feeding bottle.

4. The method of any preceding claim, wherein the milk comprises mammalian milk.

5. The method of claim 4, wherein the mammalian milk comprises human breast milk, suitably wherein the human breast milk is freshly expressed human breast milk.

6. The method of any preceding claim, wherein the OSS comprises: a. a body comprising a first end and a second end, wherein the first and second ends are open; and wherein the body defines a first interior compartment; b. a lid adapted to seal the first end of the body; c. a gas permeable, liquid impermeable membrane, said membrane being suitable for closing the second end of the body; and d. an oxygen scavenging material (OSM), said OSM being suitable for placement in the first interior compartment.

7. The method of any preceding claim wherein the OSS further comprises a gasket, said gasket suitable for providing a seal between an opening of the sealable storage container and a cap suitable for sealing the opening of the sealable storage container.

8. The method of claim 7, wherein the gasket surrounds the body and extends radially outwardly therefrom.

9. The method of claim 7 or claim 8, wherein the gasket comprises a resilient material, optionally wherein the resilient material comprises a silicone rubber.

10. The method of any one of claims 7 to 9, wherein the gasket extends radially outwardly from the body by a distance of at least 1 cm, optionally at least 2cm, suitably at least 3 cm.11 . The method of any preceding claim, wherein the gas permeable, liquid impermeable membrane is comprised of a hydrophobic material, or comprises a hydrophobic coating applied thereto.

12. The method of any preceding claim, wherein the gas permeable, liquid impermeable membrane further comprises a gas impermeable, liquid impermeable covering.

13. The method of any preceding claim, wherein the OSM is selected from one or more of: ferrous iron powder with sodium chloride; optionally with activated carbon; a ferrous carbonate in conjunction with a metal halide catalyst; ferrous iron oxide; unsaturated fatty acid; sodium hydrogen carbonate; citric acid and ascorbic acid.

14. The method of any preceding claim, wherein the amount of OSM for placement in the first interior compartment is at least 1 g, at least 2 g, suitably at least 5 g, optionally at least 10 g; and at most 30 g, suitably at most 40 g; optionally at most 50 g.

15. The method of any preceding claim, wherein the OSS further comprises a sleeve, wherein the sleeve surrounds the body of the OSS, optionally wherein the gasket (when present) surrounds the sleeve.

16. The method of any preceding claim, wherein the anaerobic microbiome comprises microorganisms selected from anaerobic bacteria, optionally selected from Bifidobacteriaceae and / or Lactobacillaceae, suitably wherein the Bifidobacteria are selected from Bifidobacterium infantis, Bifidobacterium breve, Bifidobacterium iongum, Bifidobacterium animalis and / or Bifidobacterium bifidum.

17. An oxygen scavenging system (OSS) for use with a sealable storage container to preserve the viability of an anaerobic microbiome in a medium comprised within, said OSS comprising: a) a body comprising a first end and a second end, wherein the first and second ends are open; and wherein the body defines a first interior compartment; b) a lid adapted to seal the first end of the body; c) a gas permeable, liquid impermeable membrane, said membrane being suitable for closing the second end of the body; and d) an oxygen scavenging material (OSM), said OSM being suitable for placement in the first interior compartment.

18. The OSS of claim 17 further comprising a gasket, said gasket suitable for providing a seal between an opening of the storage container and a cap suitable for closing the opening of the storage container.

19. The OSS of claim 18, wherein the gasket surrounds the body and extends radially outwardly therefrom.

20. The OSS of claim 18 or 19, wherein the gasket comprises a resilient material, optionally wherein the resilient material comprises a silicone rubber.

21. The OSS according any one of claim 17 to 20, wherein the gasket extends radially outwardly from the body by a distance of at least 0.5cm, optionally at least 1 cm, at least 1 .5 cm, at least 2cm, at least 2.5 cm, at least 3 cm, at least 3.5 cm, at least 4 cm, at least 4.5 cm, at least 5 cm, at least 5.5 cm, at least 6 cm.

22. The OSS of any one of claims 17 to 21 , wherein the gas permeable, liquid impermeable membrane is comprised of a hydrophobic material, or comprises a hydrophobic coating applied thereto.

23. The OSS of any one of claims 17 to 22, wherein the gas permeable, liquid impermeable membrane further comprises a hydrophobic coating.

24. The OSS of any one of claims 17 to 23, wherein the OSM is selected from one or more of: ferrous iron powder with sodium chloride; optionally with activated carbon; a ferrous carbonate in conjunction with a metal halide catalyst; ferrous iron oxide; unsaturated fatty acid; sodium hydrogen carbonate; citric acid and ascorbic acid.

25. The OSS of any one of claims 17 to 24, wherein the amount of OSM for placement in the first interior compartment is at least 2 g, suitably at least 5 g, optionally at least 10 g; and at most 30 g, suitably at most 40 g; optionally at most 50 g.

26. The OSS of any one of claims 17 to 25, further comprising a sleeve.

27. The OSS of claim 26, wherein the sleeve surrounds the body, optionally wherein the gasket (when present) surrounds the sleeve.

28. The OSS of claim 26 or claim 27, wherein the sleeve is frustoconical.

29. The OSS of any one of claims 17 to 28, wherein the OSM is placed in the first interior compartment, the first end is sealed by the lid and the second end is sealed by the gas permeable liquid impermeable membrane.

30. The OSS of one of claims 17 to 28, wherein the gas permeable liquid impermeable membrane is covered by a gas impermeable, liquid impermeable covering.

31. The OSS of claim 30, wherein the gas impermeable, liquid impermeable covering can be peeled off by a user.

32. The OSS of any one of claims 17 to 31 , wherein the storage container is selected from: a vessel, a bottle, a sealable bag or a pouch.

33. The OSS of claim 32 wherein the bottle is an infant feeding bottle.

34. The OSS of any one of claims 17 to 33, wherein the milk comprises mammalian milk, suitably wherein the mammalian milk is unpasteurised.

35. The OSS of any one of claims 17 to 34, wherein the anaerobic microbiome comprises microorganisms selected from anaerobic bacteria, optionally selected from Bifidobacteriaceae and / or Lactobacillaceae, suitably wherein the Bifidobacteria are selected from Bifidobacterium infantis, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium animalis and / or Bifidobacterium bifidum.

36. A method for preserving the shelf life of freshly expressed milk, said method comprising storing milk in a storage container wherein the storage container comprises an oxygen scavenging system (OSS).

37. The method of claim 36, wherein the OSS is of any one of claims 17 to 35.

38. The method of claim 36 or claim 37, wherein the milk is mammalian milk, optionally human breast milk.

39. An infant feeding bottle comprising an oxygen scavenging system (OSS).

40. The infant feeding bottle of claim 39, wherein the OSS comprises: a. a body comprising a first end and a second end, wherein the first and second ends are open; and wherein the body defines a first interior compartment; b. a lid adapted to seal the first end of the body; c. a gas permeable, liquid impermeable membrane, said membrane being suitable for closing the second end of the body; and d. an oxygen scavenging material (OSM), said OSM being suitable for placement in the first interior compartment.41 . The infant feeding bottle of claim 40, wherein the amount of OSM for placement in the first interior compartment is at least 1 g, suitably at least 2 g, suitably at least 5 g, optionally at least 10 g; and at most 30 g, suitably at most 40 g; optionally at most 50 g.

42. The infant feeding bottle of claim 40 or claim 41 , wherein the OSM is selected from one or more of: ferrous iron powder with sodium chloride; optionally with activated carbon; a ferrous carbonate in conjunction with a metal halide catalyst; ferrous iron oxide; unsaturated fatty acid; sodium hydrogen carbonate; citric acid and ascorbic acid.

43. The infant feeding bottle of any one of claims 39 to 42, wherein the gasket extends radially outwardly from the body by a distance of at least 0.5cm, optionally at least 1 cm, at least 1.5 cm, at least 2cm, at least 2.5 cm, at least 3 cm, at least 3.5 cm, at least 4 cm, at least 4.5 cm, at least 5 cm, at least 5.5 cm, at least 6 cm.

44. The infant feeding bottle of any one of claims 39 to 43, wherein the OSS selectively promotes the growth of probiotic anaerobic microorganisms and suppresses the growth of pathogenic microorganisms in a medium, optionally wherein the medium is mammalian milk, optionally human breast milk.

45. A sealable container for preserving the viability of anaerobic microorganisms comprised within, the container comprising the OSS of any of claims 17 to 35.

46. A cap for sealing a sealable container for preserving the viability of anaerobic microorganisms comprised within, wherein the cap is adapted to receive the OSS of any of claims 17 to 35.M . The OSS of any of claims 17 to 35 for preserving the antioxidant content of a medium, such as milk.

48. The OSS of any one of claim 17 to 35, wherein the OSS selectively promotes the growth of probiotic anaerobic microorganisms and suppresses the growth of pathogenic microorganisms in a medium, optionally wherein the medium is mammalian milk, optionally human breast milk.

49. The OSS of any one of claims 17 to 35, further comprising an oxygen indicator.

50. The OSS of claim 49, wherein the oxygen indicator is a dye.51 . A method for improving the growth of probiotic microorganisms or, selectively promoting the growth of probiotic anaerobic microorganisms and suppressing the growth of pathogenic microorganisms, in freshly expressed human breast milk, said method comprising storing the breastmilk in an infant feeding bottle comprising an OSS of any one of claims 17 to 35.

52. The method of claim 51 , wherein the milk comprises Bifidobacteriaceae and / or Lactobacillaceae, optionally wherein the Bifidobacteria are selected from Bifidobacterium infantis, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium animalis and / or Bifidobacterium bifidum.

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