Devices and methods for opening shells of shellfish and releasing meat from shells of shellfish through isochoric freezing

Abstract

The present disclosure provides methods and devices for opening shellfish and detaching a meat from the shells of the shellfish through isochoric freezing treatments. Also, the present disclosure provides methods and devices for partially opening the shellfish and partially detaching a meat from the shells of the shellfish, thereby retaining liquor and avoiding meat dehydration.

Description

Attorney Docket: BCHR-002WODEVICES AND METHODS FOR OPENING SHELLS OF SHELLFISH AND RELEASING MEAT FROM SHELLS OF SHELLFISH THROUGH ISOCHORIC FREEZINGCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims benefit of priority to U.S. Provisional Application No. 63 / 689,543, filed August 30, 2024, the disclosure of which is incorporated herein by reference in its entirety.TECHNICAL FIELD

[0002] The present disclosure relates to methods and devices for opening shells of shellfish and releasing meat from the shells of shellfish through treatment with isochoric freezing.INTRODUCTION

[0003] In the quest for sustainable food sources, shellfish emerges as a remarkably eco- friendly option compared to traditional animal meat. This advantage primarily stems from the energy dynamics involved in protein production. To understand why shellfish are superior in this regard, it is essential to delve into the intricacies of i) energy consumption, ii) resource use and environmental impact, and iii) nutritional and economic benefits associated with both shellfish and conventional livestock farming.

[0004] i) Energy consumption

[0005] First, shellfish, such as oysters, mussels, and clams, occupy a lower trophic level in the food web compared to land-based livestock like cows, pigs, and chickens. This positioning is crucial because energy transfer between trophic levels is inherently inefficient. According to the ten percent rule in ecology, only about 10% of the energy from one trophic level is passed on to the next. Consequently, producing animal meat, which requires feeding herbivores or omnivores with plant matter, is significantly less efficient than harvesting protein directly from lower trophic levels, such as shellfish.Attorney Docket: BCHR-002WO

[0006] Furthermore, many shellfish are filter feeders, meaning they extract their food directly from the water by filtering plankton and other small organic particles. This feeding method bypasses the need for cultivated feed, which is a substantial energy sink in conventional livestock farming. For instance, growing feed crops requires significant inputs of water, fertilizer, and energy for planting, harvesting, and transporting. Shellfish, by contrast, naturally obtain their nutrients from their aquatic environment, thus minimizing additional energy inputs.

[0007] Finally, shellfish exhibit high feed conversion ratios (FCRs), which measure the efficiency with which animals convert feed into body mass. Bivalves like oysters and mussels have FCRs close to 1:1, meaning they almost convert all the food they consume into biomass. In contrast, the FCR for cattle ranges between 6:1 to 8:1, signifying that producing one kilogram of beef requires six to eight kilograms of feed. This stark difference underscores the superior energy efficiency of shellfish in protein production.

[0008] ii) Resource Use and Environmental Impact

[0009] First, land use is a critical factor in environmental sustainability. Traditional livestock farming requires vast tracts of land for grazing or growing feed crops. This land use contributes to deforestation, habitat destruction, and soil degradation. Shellfish aquaculture, however, primarily utilizes coastal and marine environments, which do not compete with terrestrial ecosystems. Furthermore, shellfish farming often takes place on underwater structures, such as ropes and cages, making it a highly space-efficient method of food production.

[0010] Secondly, water use in agriculture is another area where shellfish have a distinct advantage. Livestock farming is notorious for its high water consumption, not only for the animals themselves but also for growing their feed. Producing a kilogram of beef, for example, can require up to 15,000 liters of freshwater. In contrast, shellfish farming generally does not require additional freshwater inputs beyond the natural aquatic environment, significantly reducing its water footprint.

[0011] Thirdly, the carbon footprint of shellfish is markedly lower than that of land- based meat. Livestock farming is a significant source of greenhouse gases, particularly methane, which is emitted by ruminants like cows. Shellfish, conversely, do not produceAttorney Docket: BCHR-002WO methane and their farming typically involves lower energy consumption for maintenance and harvesting, resulting in a smaller overall carbon footprint.

[0012] Finally, beyond merely having a lower negative impact, shellfish can actively contribute to environmental health. Filter-feeding bivalves improve water quality by removing excess nutrients and particulates from the water, which can help combat issues like eutrophication. This natural filtration service supports clearer waters and healthier aquatic ecosystems. Additionally, shellfish beds provide habitat for other marine organisms, enhancing local biodiversity.

[0013] iii) Nutritional and economic benefits

[0014] Shellfish offer high-quality protein with a favorable amino acid profile, making them a nutritious alternative to traditional meat. They are also rich in essential minerals, vitamins, and omega-3 fatty acids, contributing to a balanced diet.

[0015] Furthermore, shellfish farming is often less capital-intensive compared to livestock farming. It requires fewer inputs such as feed and water, and the infrastructure for shellfish aquaculture can be simpler and more cost-effective to establish and maintain. This economic advantage makes shellfish farming an attractive option for coastal communities, supporting livelihoods while promoting sustainable food production.

[0016] Thus, in light of the pressing need for sustainable food sources, shellfish present a compelling alternative to traditional animal meat. Their superior energy efficiency, minimal resource use, and lower environmental impact position them as a vital component of a sustainable future food system. Embracing shellfish as a significant protein source can help reduce the ecological footprint of our diets while providing high-quality nutrition and supporting economic resilience in coastal regions. As we look to feed a growing global population within the planet's ecological limits, shellfish farming offers a beacon of sustainability and hope.

[0017] However, one of the key obstacles for why more consumers have not incorporated more shellfish in their diet, which would in turn reduce the need for the environmentally unfriendly animal meat, is price. The meat from oysters, blue mussels and most other shellfish is more expensive than animal meat and if it was more affordable it wouldAttorney Docket: BCHR-002WO appeal to a larger audience, quite certainly leading to a significant increase in consumption and thus reduction of animal meat consumption and subsequently production.

[0018] Opening of the shells and removing the meat is a significant factor in the price. If a new method were developed that could open the shells and free the meat from the shells at low cost, it would lead to a decrease in the price of shellfish, with all the benefits described above.SUMMARY

[0019] Aspects of the present disclosure include methods and devices for processing shellfish by isochoric freezing. More specifically, methods and devices of the present disclosure are for detaching a meat inside shellfish. Furthermore, methods and devices of the present disclosure are for opening shellfish and detaching a meat inside the shellfish. The connective tissue of shellfish to the shells is compromised by the joint effects of subfreezing temperature and pressure under the conditions of isochoric freezing. In particular, the isochoric freezing reduces or inhibits enzymatic activity in shellfish and weakens adhesion of the shellfish to its shells. Aspects of the present disclosure further provide methods and devices for partially opening the shells of the shellfish and partially detaching a meat from the shells of the shellfish through isochoric freezing treatment, i.e. partial shucking of shellfish.

[0020] In some aspects, the present disclosure provides a method for processing shellfish, comprising a) placing shellfish in an isochoric chamber filled with a fluid; and b) isochoric freezing the fluid in the isochoric chamber containing the shellfish at a subfreezing temperature over a treatment time. In other aspects, a method for partially opening shellfish comprises a) placing shellfish in an isochoric chamber filled with a fluid, and b-2) isochoric freezing the fluid in the isochoric chamber with a subfreezing temperature of -0.1 °C or less, a pressure of 1 MPa or more, and a treatment time of 30 minutes or more. Partially opening shellfish means that the shellfish remains mostly closed but is structurally weakened, thereby retaining liquor and avoiding meat dehydration. The attachment of the meat to the shells is also weakened. In other words, partially opening shellfish includes partially detaching the meat from the shells of the shellfish. In some embodiments, the method further comprises c) warming the fluid in the isochoric chamber containing the shellfish.Attorney Docket: BCHR-002WO

[0021] In some embodiments, the treatment time for processing shellfish is about 1 hour to 96 hours. In some embodiments, the treatment time for processing shellfish is about 1 hour to 72 hours. In some embodiments, the treatment time for processing shellfish is about 1 hour to 48 hours. In other embodiments, the treatment time for processing shellfish is about 1 hour to 24 hours.

[0022] In some embodiments, the subfreezing temperature for processing shellfish is in the range of about 0 °C to -20 °C. In other embodiments, the subfreezing temperature for processing shellfish is in the range of about 0 °C to -10 °C. In still other embodiments, the subfreezing temperature for processing shellfish is in the range of about 0 °C to -5 °C. In other embodiments, the subfreezing temperature for processing shellfish is in the range of about 0 °C to -3 °C. In yet other embodiments, the subfreezing temperature for processing shellfish is in the range of about 0 °C to -2.5 °C. In yet other embodiments, the subfreezing temperature for processing shellfish is in the range of about 0 °C to -2 °C.

[0023] In some embodiments, the isochoric freezing for processing shellfish is at a pressure in the range of about 0.1 MPa to about 150 MPa. In other embodiments, the isochoric freezing for processing shellfish is at a pressure in the range of about 0.1 MPa to about 100 MPa. In still other embodiments, the isochoric freezing for processing shellfish is at a pressure in the range of about 0.1 MPa to about 55 MPa. In other embodiments, the isochoric freezing for processing shellfish is at a pressure in the range of about 0.1 MPa to about 35 MPa. In other embodiments, the isochoric freezing for processing shellfish is at a pressure in the range of about 0.1 MPa to about 30 MPa. In yet other embodiments, the isochoric freezing for processing shellfish is at a pressure in the range of about 0.1 MPa to about 25 MPa.

[0024] In some embodiments, the isochoric freezing conditions for partially opening shells of shellfish include a subfreezing temperature of -0.1 °C or less, a pressure of 1 MPa or more, and a treatment time of 30 minutes or more. In certain embodiments, the isochoric freezing conditions for partially opening the shellfish include a subfreezing temperature of about -0.1 °C to -3.5 °C, a pressure of about 1 MPa to 35 MPa and a treatment time of about 30 minutes to 72 hours. In certain embodiments, the isochoric freezing conditions for partially opening the shellfish include a subfreezing temperature of about -0.1 °C to -3.0 °C, a pressure of about 1 MPa to 30 MPa and a treatment time of about 30 minutes to 48 hours. In certainAttorney Docket: BCHR-002WO embodiments, the isochoric freezing conditions for partially opening the shellfish include a subfreezing temperature of about -1.0 °C to -3.0 °C, a pressure of about 10 MPa to 30 MPa and a treatment time of about 12 hours to 72 hours. In certain embodiments, the isochoric freezing conditions for partially opening the shellfish include a subfreezing temperature of about -2.0 °C to -3.0 °C, a pressure of about 20 MPa to 30 MPa and a treatment time of about 36 hours to 72 hours. In some embodiments, the isochoric freezing conditions for partially opening the shellfish include a subfreezing temperature of about -2.5 °C to -3.0 °C, a pressure of about 25 MPa to 30 MPa and a treatment time of about 48 hours to 72 hours. In other embodiments, the isochoric freezing conditions for partially opening the shellfish include a subfreezing temperature of about -3.0 °C, a pressure of about 30 MPa and a treatment time of about 72 hours.

[0025] In some embodiments, the method of the present disclosure is used for bulk processing. For bulk processing, a large number of shellfish can be placed in the isochoric chamber and volume of the isochoric chamber is suitable for bulk processing. In some embodiments, meat of the large number of shellfish are detached from the shells of the shellfish through the isochoric freezing. In certain embodiments, the large number of shellfish are opened through the isochoric freezing and meat of the large number of shellfish are also detached from the shells of the shellfish through the isochoric freezing.

[0026] In some embodiments, the shellfish is directly placed in the isochoric chamber. In other embodiments, the shellfish is placed in a matter container, and the matter container is placed in the fluid in the isochoric chamber.

[0027] In some embodiments, the shellfish is vacuum packed and placed in the isochoric chamber. In other embodiments, the shellfish is not vacuum packed and placed in the isochoric chamber.

[0028] In some embodiments, the shellfish is alive prior to the isochoric freezing. In some embodiments, the shellfish is raw shellfish or undercooked shellfish.In some embodiments, the shellfish is selected from a bivalve, a univalve, a crustacean, a gastropod, a scaphopod, a polyplacophoran, or an echinoderm. In certain embodiments, the bivalve is selected from a mussel, a scallop, a clam, a winkle, and an oyster. In certain embodiments, wherein the univalve is a snail. In certain embodiments, the crustacean isAttorney Docket: BCHR-002WO selected from a shrimp, a prawn, a crab, a lobster, barnacles, and a crayfish. In certain embodiments, the gastropod is selected from a conch, an abalone, a limpet, and a whelk.

[0029] In some embodiments, 30% or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In other embodiments, 50% or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In still other embodiments, 70% or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In yet other embodiments, 90% or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In further embodiments, all of the shellfish are open and the meat inside the shellfish are detached through the isochoric freezing.

[0030] In some aspects, the present disclosure provides a device for processing shellfish comprising a) an isochoric chamber filled with a fluid, wherein the isochoric chamber contains shellfish; b) a temperature control system configured to cool the isochoric chamber containing the shellfish to a subfreezing temperature over a treatment time and then warm the isochoric chamber containing the shellfish. The device for processing the shellfish enables opening the shellfish and / or detaching a meat inside the shellfish through the isochoric freezing. In some embodiments, the device is used for partially opening the shells of the shellfish, where the shellfish remains mostly closed but is structurally weakened, thereby retaining liquor and avoiding meat dehydration. The attachment of the meat to the shells is also weakened. In other words, partially opening the shells of the shellfish includes partially detaching the meat from the shells of the shellfish.

[0031] In some embodiments, the device of the present disclosure comprises a pressure monitoring element. In some embodiments, the device of the present disclosure comprises a pressure relief mechanism. In some embodiments, the device of the present disclosure comprises multiple discrete sub-systems for physically separating shellfish from ice.Attorney Docket: BCHR-002WO

[0032] In some embodiments, the treatment time for processing shellfish is 1 hour to 96 hours. In some embodiments, the treatment time for processing shellfish is 1 hour to 72 hours. In some embodiments, the treatment time for processing shellfish is 1 hour to 48 hours. In other embodiments, the treatment time for processing shellfish is from 1 hour to 24 hours.

[0033] In some embodiments, the subfreezing temperature for processing shellfish is in the range of about 0 °C to -20 °C. In other embodiments, the subfreezing temperature for processing shellfish is in the range of about 0 °C to -10 °C. In still other embodiments, the subfreezing temperature for processing shellfish is in the range of about 0 °C to -5 °C. In other embodiments, the subfreezing temperature for processing shellfish is in the range of about 0 °C to -3 °C. In other embodiments, the subfreezing temperature for processing shellfish is in the range of about 0 °C to -2.5 °C. In yet other embodiments, the subfreezing temperature for processing shellfish is in the range of about 0 °C to -2 °C.

[0034] In some embodiments, the isochoric freezing for processing shellfish is at a pressure in the range of about 0.1 MPa to about 150 MPa. In other embodiments, the isochoric freezing for processing shellfish is at a pressure in the range of about 0.1 MPa to about 100 MPa. In still other embodiments, the isochoric freezing for processing shellfish is at a pressure in the range of about 0.1 MPa to about 55 MPa. In other embodiments, the isochoric freezing for processing shellfish is at a pressure in the range of about 0.1 MPa to about 35 MPa. In other embodiments, the isochoric freezing for processing shellfish is at a pressure in the range of about 0.1 MPa to about 30 MPa. In yet other embodiments, the isochoric freezing for processing shellfish is at a pressure in the range of about 0.1 MPa to about 25 MPa.

[0035] In some embodiments, the isochoric freezing condition for partially opening shells of shellfish includes a subfreezing temperature of -0.1 °C or less, a pressure of 1 MPa or more, and a treatment time of 30 minutes or more. In certain embodiments, the isochoric freezing conditions for partially opening the shellfish include a subfreezing temperature of about -0.1 °C to -3.5 °C, a pressure of about 1 MPa to 35 MPa, and a treatment time of about 30 minutes to 72 hours. In certain embodiments, the isochoric freezing conditions for partially opening the shellfish include a subfreezing temperature of about -0.1 °C to -3.0 °C, a pressure of about 1 MPa to 30 MPa, and a treatment time of about 30 minutes to 48 hours. In certain embodiments, the isochoric freezing conditions for partially opening the shellfish include aAttorney Docket: BCHR-002WO subfreezing temperature of about -1.0 °C to -3.0 °C, a pressure of about 10 MPa to 30 MPa, and a treatment time of about 12 hours to 72 hours. In certain embodiments, the isochoric freezing conditions for partially opening the shellfish include a subfreezing temperature of about -2.0 °C to -3.0 °C, a pressure of about 20 MPa to 30 MPa, and a treatment time of about 36 hours to 72 hours. In some embodiments, the isochoric freezing conditions for partially opening the shellfish include a subfreezing temperature of about -2.5 °C to -3.0 °C, a pressure of about 25 MPa to 30 MPa, and a treatment time of about 48 hours to 72 hours. In other embodiments, the isochoric freezing conditions for partially opening the shellfish include a subfreezing temperature of about -3.0 °C, a pressure of about 30 MPa, and a treatment time of about 72 hours.

[0036] In some embodiments, the device is suitable for bulk processing. In some embodiments, the volume of the isochoric chamber is suitable for bulk processing. In some embodiments, the isochoric chamber can contain a large number of shellfish. In certain embodiments, volume of the isochoric chamber is about 50 L to 10,000 L. In certain embodiments, the volume of the isochoric chamber is about 1,000 L to 2,000 L.

[0037] In some embodiments, meat of the large number of shellfish are detached from the shellfish through the isochoric freezing. In other embodiments, the large number of shellfish are opened and meat inside the shellfish are detached through the isochoric freezing. In certain embodiments, 30% or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In certain embodiments, 50% or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In certain embodiments, 70% or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In certain embodiments, 90% or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In certain embodiments, all of the shellfish are open and the meat inside the shellfish are detached through the isochoric freezing.Attorney Docket: BCHR-002WOBRIEF DESCRIPTION OF THE FIGURES

[0038] FIG. 1 shows a temperature-pressure phase diagram for water and comparison of path of freezing in an isochoric system and an isobaric system.

[0039] FIG. 2 shows the plot of the percentage of ice during isochoric freezing, more specifically the percentage of ice in an isochoric system as the temperature is lowered to the triple point.

[0040] FIG. 3 illustrates a typical isochoric chamber for isochoric freezing.

[0041] FIG. 4 shows how biological matter can be preserved at subfreezing temperatures while avoiding the formation of ice. As shown in FIG. 4, it is possible to reduce the temperature in isochoric freezing while avoiding damage from the formation of ice by placing the matter to be preserved in the unfrozen part of the volume.

[0042] FIG. 5 shows an isochoric freezing configuration with an isochoric chamber and a matter container for biological matter preservation.

[0043] FIG. 6 shows that the isochoric system comprises an isochoric chamber that is enclosed with a sealing cap. The isochoric chamber can be made of 316 stainless steel with a total volume capacity of 2000 liters and pressure-rated for up to 110 MPa. The chamber can be connected to an electronic pressure transducer to monitor the pressure inside the chamber. A safety head with a rupture disk is in fluid communication with the interior of the isochoric chamber to ensure that the conditions inside the chamber do not exceed safety standards.

[0044] FIG. 7 a blue mussel that was subjected to isochoric freezing at -2 °C for 96 hours.

[0045] FIG. 8A shows blue mussel that has not received isochoric freezing and has been forced open, and FIG. 8B blue mussel from the same batch after it was subjected to isochoric freezing at -6 °C for 48 hours.

[0046] FIGS. 9A and 9B show an oyster that has not received isochoric freezing. FIG. 9A shows the oyster is closed. FIG. 9B shows that meat is attached to its shell when it has been forced open. FIG. 9C shows that an oyster is from the same batch after they were subjected to isochoric freezing at -7 °C for 72 hours.

[0047] FIG. 10 shows relationship between storage time and partial shucking efficiency.

[0048] FIGS. 11A-11E shows a visual documentation of partial shucking.Attorney Docket: BCHR-002WODETAILED DESCRIPTION

[0049] As a compelling alternative to traditional animal meat, shellfish emerges as a remarkably eco-friendly option compared to traditional animal meat. Their superior energy efficiency, minimal resource use, and lower environmental impact position them as a vital component of a sustainable future food system. However, the meat from oysters, blue mussels and most other shellfish is more expensive than animal meat and if it was more affordable it would appeal to a larger audience, quite certainly leading to a significant increase in consumption. In particular, opening of the shells and removing the meat is a significant factor in the price.

[0050] To achieve price reduction for processing shellfish, the present disclosure provides a method or device for opening shells of shellfish and detaching a meat inside of the shellfish through isochoric freezing with several advantages as follows:

[0051] i) Reduced Labor Costs: Eliminating the labor costs associated with shucking would lower the overall production costs for shellfish. Producers could pass these savings on to consumers in the form of lower prices.

[0052] ii) Increased Efficiency: A new, low-cost method for processing shellfish of the present disclosure would likely be faster and more efficient than manual labor, increasing the supply of shellfish available to the market. Greater supply often leads to lower prices.

[0053] iii) Broader Accessibility: Lower prices would make shellfish more accessible to a wider range of consumers who might previously have considered them a luxury item.

[0054] iv) Market Expansion: Restaurants and food processors might be more inclined to include shellfish in their offerings if the cost and labor involved in preparation were reduced, further driving demand.

[0055] v) Consumer Perception: The convenience of pre-shucked shellfish could attract more consumers, including those who may have been deterred by the effort required to prepare them.

[0056] Regarding shellfish processing, US2008030523A1 discloses High Pressure Processing (HPP) systems. The HPP systems are used to open up shells of shellfish and releaseAttorney Docket: BCHR-002WO meat from their shell through the mechanisms, i.e. by having very high pressure denature the proteins in the connective tissue between shell and meat in the shellfish. The HPP system is performed at temperatures above 0 °C as lower temperatures decrease efficiency.

[0057] In contrast, a method or device of the present disclosure is always performed below 0°C as isochoric freezing is applied to release meat from shells of shellfish. As such, lower temperature increases the efficiency of the release in the present disclosure in contrast to the HPP technology where lower temperatures decrease efficiency.

[0058] Furthermore, the present disclosure provides that the combined effects of low temperature and low pressure disrupt the connective tissue between shell and the meat in shellfish, weaken enzymatic activities, and cause phase transitions in lipids. This happens without the need to denature proteins, which is the prerequisite for release of shellfish from shell with HPP technology (e.g. pressure in HPP is not applied below 100 MPa and generally above 200 MPa). The low pressure of 26 MPa applied in Example 1 for the present invention is not sufficient to cause the denaturing of actin and myosin and connective tissues to a gelatin transition at temperatures above 0 °C.

[0059] Thus, the present invention has distinct features from the known HPP technology in that meat is detached from shells of shellfish i) at pressures below what is needed to denature proteins (while denaturing of proteins is a prerequisite for release in HPP), ii) only at temperatures below 0 °C (while HPP is performed above 0°C) iii) by applying treatment times that is at least 1 hour (while no HPP processes are described to be 1 hour or more), and iv) it shows reverse level of efficiency changes resulting from temperature increase / decrease compared to HPP.

[0060] In particular, certain pressures, significantly lower than those used in high- pressure processing, can lead to partial shell opening, enabling faster shucking with a manual or mechanical method in the next step, and thereby increasing the efficiency of the overall process.

[0061] The present disclosed subject matter now will be described more fully hereinafter with reference to the accompanying Figures, in which some, but not all embodiments of the inventions are shown. Like numbers refer to like elements throughout.Attorney Docket: BCHR-002WOThe presently disclosed subject matter may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. Indeed, many modifications and other embodiments of the presently disclosed subject matter set forth herein will come to mind to one skilled in the art to which the presently disclosed subject matter pertains having the benefit of the teachings presented in the foregoing descriptions and the associated Figures. Therefore, it is to be understood that the presently disclosed subject matter is not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims.

[0062] Section headings as used in this section and the entire disclosure herein are merely for organizational purposes and are not intended to be limiting.1. Definitions

[0063] Before the present invention is further described, it is to be understood that this invention is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0064] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0065] Unless defined otherwise, 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. Although any methods and materials similar or equivalent to thoseAttorney Docket: BCHR-002WO described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited.

[0066] As used herein, the term "isochoric freezing" refers to the freezing of aqueous solutions in a constant volume chamber or in a rigid chamber. In isochoric freezing, the phase transition encompasses the entire volume when cooling below the liquidus line and a substantial portion of the volume remains unfrozen to the triple point temperatures and pressures and is associated with an increase in pressure.

[0067] As used herein, the term "isobaric freezing" refers to a freezing process at constant pressure. In isobaric freezing, the entire system freezes as soon as the temperature is below the liquidus line intersection with the constant pressure line.

[0068] As used herein, the term "shellfish" refers to a shelled mollusk (e.g. an oyster or cockle), a crustacean (e.g. a crab or shrimp), a bivalve, a univalve, a gastropod, a scaphopod, a polyplacophora, and an echinoderm, especially one that is edible. Herein, the present disclosure applies to all types and species of shellfish. As such, shellfish includes anything from clams and oysters to lobster and shrimp and includes edible freshwater invertebrates such as crayfish and river mussels.

[0069] As used herein, the term "shelled mollusk" or "mollusk" refers to any of a large phylum (Mollusca) of invertebrate animal (such as snails, clams, or squids) with a soft unsegmented body usually enclosed in a calcareous shell. For example, a shelled mollusk used as a food source by humans includes, but not limited to, a clam, a mussel, an oyster, a winkle, and a scallop.

[0070] As used herein, the term "bivalve" refers to any of more than 15,000 species of clams, oysters, mussels, cockles, scallops, and other members of the phylum Mollusca characterized by a shell that is divided from front to back into left and right valves. The valves are connected to one another at a hinge. As such, the bivalve lives in saltwater, as well as in freshwater. For example, bivalve includes, but is not limited to, a clam, a blood clam, a mactra chinensis, a unionida, a meretrix lusoria, a heterodonta, a soft-shell calm, an ark clam, an Atlantic surf clam, a razor claim, Atlantic jackknife clam, an oyster, a true oyster, an akoya pearlAttorney Docket: BCHR-002WO oyster, a mussel, a freshwater pearl mussel, a scallop, a pinna nobillis, a cockle, pinna nobilis, a warty venus, and a mya.

[0071] As used herein, the term "univalve" refers to univalve is a mollusk with a single shell, such as a snail (land dwelling mollusks) or periwinkle (ocean dwelling mollusks). Gastropods, a type of mollusk, are sometimes called univalves because the shell of most gastropods is usually one piece. For example, univalve includes, but is not limited to, a burgundy snail, a giant African snail, a cerithidea obtusa, an apple snail, a red-rimmed melania, a cone snail, a helix, a patella vulgate, a milk snail.

[0072] The term "crustacean" refers to an invertebrate animal that constitutes one group of arthropods that are a part of the subphylum Crustacea, a large, diverse group of mainly aquatic arthropods including decapods (shrimps, prawns, crabs, lobsters and crayfish), seed shrimp, branchiopods, fish lice, krill, remipedes, isopods, barnacles, copepods, opossum shrimps, amphipods and mantis shrimp. In certain embodiments, crustacean is, but not limited to, a shrimp, a prawn, a lobster, a crab, a barnacle, an isopod, a decapod, a woodlouse, or crayfish. For example, a shrimp includes, but is not limited to, a caridean shrimp, a mantis shrimp, a brine shrimp, a whiteleg shrimp, a prawn, and a krill. For example, a lobster includes, but is not limited to, a spiny lobster, slipper lobster. For example, a crab includes, but is not limited to, a coconut crab, a king crab, a red king crab, a mole crab, a true crab, or a hermit crab.

[0073] The term "gastropod" refers to any member comprising snails and slugs from saltwater, freshwater, and from the land. There are many thousands of species of sea snails and slugs, as well as freshwater snails, freshwater limpets, land snails and slugs. Gastropod is made up of the snails, which have a shell into which the animal can generally withdraw, and the slugs, which are snails whose shells have been reduced to an internal fragment or completely lost in the course of evolution. Gastropods lives in the ocean, fresh waters, and land. For example, a gastropod includes, but is not limited to, a conch, an abalone, a limpet, and a whelk, which are used as food.

[0074] As used herein, the "scaphopoda" refers to a member of a class of shelled marine mollusc with worldwide distribution and is the only class of exclusively infaunal marine molluscs. The scaphopoda are a distinctive group of molluscs commonly known as the "tuskAttorney Docket: BCHR-002WO shells" because their shells are conical and slightly curved to the dorsal side, making the shells look like tiny tusks. For example, scaphopoda includes, but is not limited to, a dentalium, a fissidentalium, a gadilida, a gadila, and a gadilida.

[0075] As used herein, the term "polyplacophora" is a group of mollusks commonly referred to as chiton. Chiton typically have a flattened oval shaped body with 8 shell plates running along their back. They are found world-wide in marine habitats from the rocky intertidal zone to the deep ocean.

[0076] As used herein, the term "echinoderm" refers to any deuterostomal animal of the phylum Echinodermata, which includes starfish, brittle stars, sea urchins, sand dollars and sea cucumbers, as well as the sessile sea lilies.

[0077] As used herein, the description "opening shells of shellfish" and "opening shellfish", which are interchangeably used, is primarily relevant to the type of shellfish that have shells that protect all the meat of the animal at once inside the shell, such as blue mussels and oysters. Shellfish such as crabs, lobsters and shrimp, where all the meat is not packed into one whole shell, the application of "opening (or opening up) shells of shellfish and releasing meat from the shells of shellfish" refers to the separation of any shell protecting any meat of the animal as a whole. Thus, the opening of shells and release of shellfish meat applies to all types of species of shellfish.

[0078] As used herein, the term "detach (detached, or detaching)", "release (releasing, or released)", and "free" are interchangeably used. The "detaching meat from shellfish" or "releasing meat from shellfish" means that meat of shellfish is removed from its shell.

[0079] As used herein, the term "nutrients" refers to substances that provide energy and physical components to the organism, allowing it to survive, grow, and reproduce.Nutrients can be basic elements or complex macromolecules and can be found in any organic matter.

[0080] The use of the terms "a," "an," and "the," and similar referents in the context of describing the disclosure (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms "comprising," "having," "including," and "containing" are to be construed as open-ended terms ( / .e., meaning "including, but not limited to,") unlessAttorney Docket: BCHR-002WO otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. For example, if the range 10-15 is disclosed, then 11, 12, 13, and 14 are also disclosed. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "such as") provided herein, is intended merely to better illuminate the embodiments of the disclosure and does not pose a limitation on the scope of the disclosure unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the embodiments of the disclosure.

[0081] As used herein, the term "about" used in connection with an amount indicates that the amount can vary by 10 % of the stated amount. For example, "about 100" means an amount of from 90-110. Where about is used in the context of a range, the "about" used in reference to the lower amount of the range means that the lower amount includes an amount that is 10% lower than the lower amount of the range, and "about" used in reference to the higher amount of the range means that the higher amount includes an amount 10 % higher than the higher amount of the range. For example, from about 100 to about 1000 means that the range extends from 90 to 1100.

[0082] The term "and / or" as used herein a phrase such as "A and / or B" is intended to include both A and B; A or B; A (alone); and B (alone). Likewise, the term "and / or" as used herein a phrase such as "A, B, and / or C" is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0083] It is understood that aspects and embodiments of the present disclosure described herein include "comprising," "consisting," and "consisting essentially of" aspects and embodiments.

[0084] It is appreciated that certain features of the present disclosure, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of this disclosure, whichAttorney Docket: BCHR-002WO are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination. All combinations of the embodiments pertaining to this disclosure are specifically embraced by the present disclosure and are disclosed herein just as if each and every combination was individually and explicitly disclosed. In addition, all sub-combinations of the various embodiments and elements thereof are also specifically embraced by the present disclosure and are disclosed herein just as if each and every such sub-combination was individually and explicitly disclosed herein.

[0085] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.2. Mechanism of action that opens shells and detaches meat from shells of shellfish

[0086] In some aspects, the present disclosure provides a method for processing shellfish by incubating under isochoric conditions. More specifically, the present disclosure provides a method for releasing or detaching meat from shellfish by incubating under isochoric conditions. Furthermore, the present disclosure provides a method for opening shells of shellfish and releasing meat from the shells by incubating under isochoric conditions.

[0087] Aspects of the present disclosure further provide methods for partially opening shells of shellfish and partially releasing meat from the shells of shellfish through treatment with isochoric freezing, i.e. partial shucking of shellfish. In some aspects of the present disclosure, certain pressures, significantly lower than those used in high-pressure processing, can lead to partial shell opening, enabling faster shucking with a manual or mechanical method in the next step, and thereby increasing the efficiency of the overall process. The term "partial shucking" refers to a partial shell opening, where the shellfish remains mostly closed but is structurally weakened. Also, the attachment of the meat to the shells is weakened. This results in easier shucking of the shellfish, enabling savings in time and money. In other words, the present disclosure provides methods that ensure that the shells remain closed post-process toAttorney Docket: BCHR-002WO retain the liquor and avoid meat dehydration, which significantly increases the quality of the shellfish's meat.

[0088] In some embodiments, the method of the present disclosure comprises a) placing shellfish in an isochoric chamber filled with a fluid and b) isochoric freezing the fluid in the isochoric chamber containing the shellfish at a subfreezing temperature over a treatment time. Specifically, the "b) isochoric freezing the fluid in the isochoric chamber containing shellfish" means that applying isochoric freezing cycle to the fluid in the isochoric chamber by lowering the temperature of the isochoric chamber system below 0 °C and thereby initiating formation of ice, thereby facilitating the opening of the shells and release of the meat from the shell of the shellfish through the conditions of low temperature and low pressure created by isochoric freezing.

[0089] The key mechanisms of action of how the subfreezing temperature with the aid of pressure open the shells and release the meat from the shells are the following:

[0090] A) Mechanical Properties of Tissue: The subfreezing temperature in the chamber alters the mechanical properties of the connective tissues in shellfish, i.e. these tissues become more vulnerable and brittle. With the addition of only low pressure to these brittle bonds, they fracture more easily. As seen in Example 1 of the present disclosure, this combination at -2 °C and only 26 MPa is sufficient to break the tissues and other bonds between the shellfish and its shell. Denaturation of proteins from pressure is not necessary as is the case in HPP technologies, as 26 MPa is not sufficient alone to denature proteins at temperatures above 0°C.

[0091] B) Reduced Enzymatic Activity: Many enzymes that maintain the structural integrity of the shellfish's attachment to the shell have reduced activity at low temperatures. This reduction in enzymatic activity weakens the adhesion of the shellfish to its shell, making it easier to release. Pressures up to 30 MPa at isochoric conditions do not denature proteins in general, so the cold temperature is seen as the primary contributor in reducing enzymatic activity, e.g. in Example 1 of the present disclosure where the pressure was below 30 MPa.

[0092] C) Phase Transition of Lipids: Lipids in the shellfish's connective tissues can undergo phase transitions at low temperatures, becoming more solid or crystalline and therefore more brittle (but without freezing). Even here by applying only low pressure to theseAttorney Docket: BCHR-002WO weakened attachments between the meat and shell, leads to the release of the meat as seen at -2 °C and low pressure of 26 MPa in Example 1 of the present disclosure.

[0093] This has been demonstrated by subjecting blue mussels to isochoric freezing at -2°C for 96 hours and -6 °C for 48 hours. The shells were completely closed and the meat that was completely stuck to the shell, before subjected to isochoric freezing. After the isochoric freezing, the shells were open and the meat completely free from the shell (FIG. 7, FIGs. 8A, 8B)

[0094] Another example is that oysters were subjected to isochoric freezing at -7°C for 72 hours. The shells were completely closed and impossible to open by hand, and the meat inside was strongly connected to the shell in one place, before subjected to isochoric freezing. After the isochoric freezing, the shells were open and the meat completely free from the shell (FIGS. 9A, 9B, 9C)

[0095] In summary, the combined effects of low temperature and mild pressure disrupt the connective tissue between shell and the meat in shellfish, weaken enzymatic activities, and cause phase transitions in lipids. This all contributes to the efficient release of shellfish from its shell, without denaturation of proteins as seen from the low pressure applied in Example 1 of the present disclosure.3. Methods for processing shellfish

[0096] Aspects of the present disclosure provide a method for processing shellfish by isochoric freezing. More specifically, a method of the present disclosure is for detaching or releasing a meat inside shellfish. Furthermore, a method of the present disclosure is for opening shellfish and detaching a meat inside the shellfish. The connective tissue of shellfish to the shells is compromised by the joint effects of subfreezing temperature and pressure under the conditions of isochoric freezing. In particular, the isochoric freezing reduces or inhibits enzymatic activity in shellfish and weakens adhesion of the shellfish to its shells. The separation of meat from shellfish results both in the shells opening up on their own as the connective tissue is no longer holding the shell clamped together, as well as easy removal of meat from shell.Attorney Docket: BCHR-002WO

[0097] In some embodiments, the present disclosure provides a method processing shellfish comprising a) placing shellfish in an isochoric chamber filled with a fluid; and b) isochoric freezing the fluid in the isochoric chamber containing shellfish at a subfreezing temperature over a treatment time. In some embodiments, the method further comprises c) warming the fluid in the isochoric chamber containing the shellfish.Isochoric freezing

[0098] Herein, the isochoric freezing refers to the freezing of aqueous solutions in a constant volume chamber. The term "constant volume chamber" is used interchangeably herein with "isochoric chamber" or "rigid chamber". In certain embodiments, the isochoric chamber can be made of grade 7075 aluminum and pressure-rated for up to 110 MPa. In certain embodiments, the chamber can be connected to an electronic pressure transducer to monitor the pressure inside the chamber. The isochoric freezing technique is described in US2007 / 0042337A1 and PCT / US24 / 33896, which are incorporated by reference herein.

[0099] In the isochoric freezing system of the present disclosure, one or more shellfish are placed inside an isochoric chamber filled with an aqueous solution. In the isochoric freezing, a substantial portion of the volume remains unfrozen which is associated with an increase in pressure. The process of isochoric freezing occurs along the liquidus line in a temperature-pressure phase diagram as the temperature is dropped because the volume in an isochoric chamber is constant.

[0100] FIG. 1 illustrates the difference between isobaric freezing and isochoric freezing in a temperature-pressure phase diagram for water. It shows the triple point for water after which at lower temperatures, there is no liquid water. It also shows that at constant pressure, as soon as the temperature is below the liquidus line intersection with the constant pressure line, the entire system freezes. In contrast, in an isochoric system, pressure and temperature are interrelated through the liquidus curve in the phase diagram of the aqueous solution. Consequently, only part of the water can freeze to become ice at thermodynamic equilibrium under the pressure and constant volume conditions. This avoids the formation of ice crystals inside the biological matter if the biological matter remains in the unfrozen region. Therefore, isochoric freezing relies on pressure to avoid ice formation inside the biological matter atAttorney Docket: BCHR-002WO subfreezing temperatures. See, e.g., Rubinsky, B., Perez, P.A. and Carlson, M.E., "The thermodynamic principles of isochoric cryopreservation", 2005, Cryobiology, 50(2), pp.121-138 herein incorporated by reference in their entireties.

[0101] In the method of the present disclosure, isochoric freezing relies on Le Chatelier's principle, which states that as water expands upon freezing within a constant volume chamber (isochoric chamber), the enhanced pressures generated hinder further formation of ice. Therefore, the freezing process in an isochoric system follows the liquidus curve in the phase diagram for water depicted in FIG. 1. This minimizes the pressure for the given temperature and its potential harmful effects, such as impacting physicochemical and nutritional properties. In some embodiments, Le Chatelier's principle also allows the transformation of an aqueous liquid solution into two coexisting phases, a solid ice phase and a liquid phase, after lowering the temperature of the liquid solution to subfreezing temperatures. This controlled freezing process allows shellfish to be placed within the liquid region of the chamber, preventing ice crystal formation and the according biophysical injury. Therefore, in some embodiments, an isochoric freezing can help retain the freshness of the shellfish without substantially impacting their physicochemical and nutritional properties.

[0102] FIG. 2 shows that a substantial percentage of the volume in a liquid state to the triple point and the percentage of ice in an isochoric system as the temperature is lowered to the triple point. Therefore, in isochoric freezing it is possible to reduce the temperature while avoiding damage from the formation of ice by placing the shellfish in the unfrozen part of the volume.

[0103] In some embodiments, the shellfish is placed inside the constant volume chamber filled with an aqueous solution in isochoric freezing. The constant volume chamber is filled with a fluid in such a way as to minimize the amount of air in the chamber. For example, FIG. 3 illustrates an exemplary isochoric chamber system for isochoric freezing. In FIG. 3, the isochoric chamber system comprises a constant volume chamber in pressure vessel 301, a pressure gauge 302, and a rupture disk 303. A safety head with a rupture disk is in fluid communication with the interior of the isochoric chamber to ensure that the conditions inside the chamber do not exceed safety standards. The constant volume chamber in pressure vessel 301 is seen in cross-sectional view 304. The constant volume chamber in pressure vessel 301 isAttorney Docket: BCHR-002WO preferably hermetically sealed, and the pressure therein is monitored with pressure gage 302. Optionally, the constant volume chamber in pressure vessel 301 can be made of stainless steel, but the present invention is not limited. In some embodiments, the isochoric chamber can be also a closed chamber with rigid walls. The constant volume chamber in pressure vessel 301 is filled with fluid. In some embodiments, the fluid in the constant volume chamber is aqueous solution or pure water. In the isochoric freezing, if ice nucleates in the constant volume chamber, the pressure of the system increases, and the nucleation can be detected by the pressure gage. For example, U.S. Patent Publication No. 2007 / 0042337A1 and PCT / US24 / 33896, incorporated by reference herein, which discusses isochoric freezing system. As one example, FIG. 6 depicts mussels 601, 603 are placed in the isochoric chamber. a) Placing shellfish in an isochoric chamber filled with a fluid

[0104] In some embodiments, the method for processing shellfish comprises a) placing shellfish in an isochoric chamber filled with a fluid. In some embodiments, one or more shellfish can be placed in the isochoric chamber. For example, the number of shellfish for processing is at least 1, at least 10, at least 100, at least 1,000, at least 10,000, at least 100,000, at least 1,000,000, or at least 10,000,000.

[0105] In some embodiments, shellfish has one shell. In other embodiments, shellfish has two or more shells. In certain embodiments, shellfish having one shell may not need to be open for processing, such as abalone. In certain embodiments, shellfish having two or more shells may need to be open for processing, such as mussels.

[0106] In some embodiments, the shellfish can be different kinds of shellfish. In other embodiments, the shellfish can be the same kind of shellfish.

[0107] In some embodiments, the method comprises placing shellfish having one shell in an isochoric chamber filled with a fluid. In other embodiments, the method comprises placing shellfish having two or more shells in an isochoric chamber filled with a fluid.

[0108] In some embodiments, the method comprises placing different kinds of shellfish in an isochoric chamber filled with a fluid. In other embodiments, the method comprises placing the same kind of shellfish in an isochoric chamber filled with a fluid.Attorney Docket: BCHR-002WO

[0109] In some embodiments, the shellfish of the present disclosure refers to a shelled mollusk, such as a clam, a snail, an oyster as well as some less familiar animals, like a tusk shell and a chiton. In some embodiments, mollusks used as a food source by humans include, but not limited to, many species of clams, mussels, oysters, winkles, and scallops. More specifically, the shellfish of the present disclosure is selected from a bivalve, a univalve, a crustacean, a gastropod, a scaphopod, a polyplacophoran, or an echinoderm. In some embodiments, the shellfish of the present disclosure is edible. As such, the present disclosure applies to all types and species of shellfish.

[0110] In certain embodiments, the bivalve is selected from a mussel, a scallop, a clam, a winkle, and an oyster. For example, a mussel is, but not limited to, a blue mussel.

[0111] In certain embodiments, the univalve is a snail. For example, univalve includes, but is not limited to, a burgundy snail, a giant African snail, a cerithidea obtusa, an apple snail, a red-rimmed melania, a cone snail, a helix, a patella vulgate, a milk snail.

[0112] In certain embodiments, the crustacean is selected from a shrimp, a prawn, a crab, a lobster, barnacles, and crayfish. Also, the crustacean is an isopod, a decapod, or a woodlouse. For example, the shrimp is, but not limited to, a caridean shrimp, a mantis shrimp, a brine shrimp, a whiteleg shrimp, a prawn, and a krill. For example, the lobster is, but not limited to, a spiny lobster, slipper lobster. For example, the crab is, but is not limited to, a coconut crab, a king crab, a red king crab, a mole crab, a true crab or a hermit crab.

[0113] In certain embodiments, the gastropod is, but not limited to, a conch, an abalone, a limpet, and a whelk.

[0114] In certain embodiments, the scaphopoda includes, but is not limited to, a dentalium, a fissidentalium, a gadilida, a gadila, and a gadilida.

[0115] In certain embodiments, echinoderm includes, but is not limited to, starfish, brittle stars, sea urchins, sand dollars and sea cucumbers, as well as the sessile sea lilies.

[0116] In some embodiments, the shellfish is alive prior to the isochoric freezing. In other embodiments, the shellfish is raw shellfish or undercooked shellfish.

[0117] In some embodiments, the method of the present disclosure is suitable for bulk processing. For bulk processing, a large number of shellfish can be placed in the isochoric chamber and volume of the isochoric chamber is suitable for bulk processing. For example, theAttorney Docket: BCHR-002WO large number of shellfish is at least 50, at least 100, at least 1,000, at least 10,000, at least 100,000, or at least 1,000,000.

[0118] In some embodiments, meat of the large number of shellfish are detached from shells of the shellfish through the isochoric freezing during bulk processing. In certain embodiments, the large number of shellfish having shells are opened through the isochoric freezing and meat of the large number of shellfish are also detached from the two or more shells of the shellfish through the isochoric freezing during bulk processing.

[0119] In some embodiments, 30 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In other embodiments, 40 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In still other embodiments, 50 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In yet other embodiments, 60 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In further embodiments, 70 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In still other embodiments, 80 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In yet other embodiments, 90 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In further embodiments, all of the shellfish are open and the meat inside the shellfish are detached through the isochoric freezing.

[0120] In some embodiments, the shellfish can be directly placed in the isochoric chamber. FIG. 4 illustrates an exemplary embodiment of isochoric freezing configuration with an isochoric chamber for a biological mass while avoiding the formation of ice. In other embodiments, a biological matter can be placed in a container that allows transfer of pressureAttorney Docket: BCHR-002WO mass and heat. In still other embodiments, a biological matter can be placed in a container that allows transfer of only pressure and heat but not mass.

[0121] In other embodiments, the shellfish is placed in a matter container, and the matter container is placed in the fluid in the isochoric chamber. FIG. 5 illustrates an exemplary embodiment of isochoric freezing configuration with an isochoric chamber and a matter container for including a biological mass while avoiding the formation of ice. Also, FIG. 5 illustrates an exemplary embodiment of isochoric freezing configuration with an isochoric chamber and a matter container for including a biological material while avoiding the formation of ice. The matter container transfer pressure but not mass in the isochoric chamber. In some embodiments, the matter container includes a biological material. In other embodiments, the matter container including a biological material is filled with a fluid that does not freeze at the isochoric temperature. Herein, the biological material can be shellfish.

[0122] In some embodiments, shellfish is vacuum packed and placed in the isochoric chamber. For example, shellfish is collected in a polyethylene sterile vacuum bag and vacuum is then created within the bag and placed inside the isochoric chamber. In other embodiments, shellfish is collected in a polyethylene sterile bag. The bag is then sealed with negligible headspace and placed inside the isochoric chamber. In other embodiments, shellfish is not vacuum packed and is placed directly in a fluid in an isochoric chamber.

[0123] In some embodiments, the fluid in the constant volume chamber is aqueous solution or pure water with or without organic molecules therein. In some embodiments, the aqueous solution that is primarily water may contain other chemicals so that the freezing point of the water-based solution may be modified as required for a specific application. For example, the water-based solution may contain salt. In certain embodiments, the external solution is water. In some embodiments, the fluid in the container can be the same as the fluid in the isochoric chamber surrounding the container. In other embodiments, the fluid in the matter container can be different from the fluid in the isochoric chamber surrounding the matter container. In other words, the fluid inside the matter container and the fluid outside the matter container can be of different types in osmotic equilibrium with the preserved biological matter. In some embodiments the isochoric chamber is filled with a fluid that does not freeze at the storage temperature, such as a solution of glycerol.Attorney Docket: BCHR-002WO

[0124] In some embodiments, the isochoric chamber may contain a nucleating agent. The nucleating agent forms an ice crystal thereby the shellfish can be stored in the unfrozen part of the volume. The ice crystal formed by the nucleating agent generates pressures of the isochoric chamber, and the enhanced pressures hinder further formation of ice. In some embodiments, the nucleating agent can be a structural element of the isochoric freezing device. For example, the nucleating agent can be walls of the isochoric chamber itself, or walls of the matter container itself. In other embodiments, the nucleating agent can be any agent that promotes the formation of ice crystal in the isochoric chamber. For example, the ice nucleating agent is, but not limited to, minute solid particles, such as dust or food particles, large molecules, ice-nucleating proteins. In some embodiments, the nucleating agent is placed in the bottom of the isochoric chamber, and the chamber is completely filled with an aqueous solution. In some embodiments, shellfish is placed in a container in an isochoric chamber that allows transfer of only pressure and heat but not mass and the isochoric chamber is filled with a fluid and nucleation agents that cause the fluid to freeze before the shellfish in the container freezes. b) Isochoric freezing the fluid in the isochoric chamber containing the shellfish at a subfreezinq temperature over a treatment time

[0125] In some embodiments, the method for processing shellfish comprises b) isochoric freezing the fluid in the isochoric chamber containing the shellfish at a subfreezing temperature over a treatment time. After the chamber is prepared and loaded, the chamber is cooled to subfreezing temperatures described in the present disclosure, preferably in a conventional freezer.

[0126] In some embodiments of the method, treatment time refers to the time applied for isochoric freezing. In other embodiments, the time applied for isochoric freezing is the time for treating shellfish in an isochoric chamber. In some embodiments, treatment time of the present disclosure refers to a length of time from minutes to hours. In other embodiments, treatment time of the present disclosure refers to a length of time from hours to days. In some embodiments, the treatment time for isochoric freezing is not limited as long as applying isochoric freezing conditions to a shellfish product in an isochoric chamber.Attorney Docket: BCHR-002WO

[0127] In certain embodiments, the treatment time for isochoric freezing is, but not limited to, 10, 15, 20, 25, 30, 40, or 50 days. In certain embodiments, the treatment time for isochoric freezing is 1 hour or more, 4 hours or more, 12 hours or more, 16 hours or more, 20 hours or more, or 24 hours or more. In some embodiments, treatment time is, but not limited to about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 75 hours, 78 hours, 80 hours, 83 hours, 85 hours, 87 hours, 90 hours, 93 hours, or 96 hours. In certain embodiments, the treatment time for isochoric freezing is 3 hours or less. In certain embodiments, the treatment time for isochoric freezing is 4 hours or less. In certain embodiments, the treatment time for isochoric freezing is 5 hours or less. In certain embodiments, the treatment time for isochoric freezing is 6 hours or less.

[0128] In some embodiments, treatment time is in the range of about 1 hour to 50 days. In certain embodiments, the treatment time for isochoric freezing is in the range of about 1 hour to 2 hours, about 1 hour to 3 hours, about 1 hour to 4 hours, about 1 hour to 5 hours, about 1 hour to 6 hours, about 1 hour to 7 hours, about 1 hour to 8 hours, about 1 hour to 9 hours, about 1 hour to 10 hours, about 1 hour to 11 hours, about 1 hour to 12 hours, about 1 hour to 13 hours, about 1 hour to 14 hours, about 1 hour to 15 hours, about 1 hour to 16 hours, about 1 hour to 17 hours, about 1 hour to 18 hours, about 1 hour to 19 hours, about 1 hour to 20 hours, about 1 hour to 21 hours, about 1 hour to 22 hours, about 1 hour to 23 hours, about 1 hour to 24 hours, 1 hour to 48 hours, 1 hour to 72 hours, or 1 hour to 96 hours. In certain embodiments, the treatment time for isochoric freezing is about 48 hours. In certain embodiments, the treatment time for isochoric freezing is about 72 hours. In certain embodiments, the treatment time for isochoric freezing is about 96 hours.Attorney Docket: BCHR-002WO

[0129] In some aspects of the method, the subfreezing temperature for processing shellfish is much higher than the triple point along the liquidus line in a temperature pressure phase diagram depicted in FIG. 1. The triple point in the temperature pressure phase diagram depicted in FIG. 1 is about -21 °C. In some embodiments, the subfreezing temperature of the present disclosure is at least 5 °C higher than the triple point along the liquidus line in a temperature pressure phase diagram depicted in FIG. 1. In other embodiments, the subfreezing temperature of the present disclosure is at least 10 °C higher than the triple point along the liquidus line in a temperature pressure phase diagram depicted in FIG. 1. In some embodiments, the subfreezing temperature of the present disclosure is in the range of about 0 °C to -20 °C, about 0 °C to -15 °C, about 0 °C to -10 °C, about 0 °C to -5 °C, about 0 °C to -3 °C, about 0 °C to -2.5 °C, about 0 °C to -2 °C. In some embodiments, the subfreezing temperature of the present disclosure is in the range of about 0 °C to -20 °C, 0 °C to -15 °C, 0 °C to -14 °C, 0 °C to -13 °C, 0 °C to -12 °C, 0 °C to -11 °C, 0 °C to -10 °C, 0 °C to -9 °C, 0 °C to -8 °C, 0 °C to -7 °C, 0 °C to -6 °C, 0 °C to -5 °C, 0 °C to -3 °C, 0 °C to -2.5 °C, or 0 °C to -2 °C. In some embodiments, the subfreezing temperature of the present disclosure is about -1 °C, -2 °C, -2.5 °C, -3 °C, -4 °C, -5 °C, -6 °C, -7 °C, -8 °C, -9 °C, -10 °C, -11 °C, -12 °C, -13 °C, or -14 °C.

[0130] In some embodiments of the method, the isochoric freezing conditions combining the above-described subfreezing temperature and the pressure which occurs at the above-described subfreezing temperature when the closed isochoric chamber is in a state of thermodynamic equilibrium is effective in opening shells of shellfish and releasing the meat inside. Furthermore, the pressures induced by the isochoric freezing of the present disclosure have further effects on avoiding ice formation inside the shellfish at the subfreezing temperature. In the method of the present disclosure, the applied pressure occurs at the above-described subfreezing temperature when the closed isochoric chamber is in a state of thermodynamic equilibrium. Therefore, the isochoric system of the present disclosure provides convenient ways to achieve subfreezing temperatures and elevated pressures only by controlling the temperature without the need for mechanical means to elevated pressure.

[0131] In some embodiments, the isochoric freezing of the present disclosure is at a pressure in the range of about 0.1 MPa to 150 MPa, about 0.1 MPa to 145 MPa, about 0.1 MPa to 140 MPa, about 0.1 MPa to 135 MPa, about 0.1 MPa to 130 MPa, about 0.1 MPa to 125Attorney Docket: BCHR-002WOMPa, about 0.1 MPa to 120 MPa, about 0.1 MPa to 115 MPa, about 0.1 MPa to 110 MPa, about 0.1 MPa to 105 MPa, about 0.1 MPa to 100 MPa, about 0.1 MPa to 95 MPa, about 0.1 MPa to 90 MPa, about 0.1 MPa to 85 MPa, about 0.1 MPa to 80 MPa, about 0.1 MPa to 75 MPa, about 0.1 MPa to 70 MPa, about 0.1 MPa to 65 MPa, about 0.1 MPa to 60 MPa, about 0.1 MPa to 55 MPa, about 0.1 MPa to 50 MPa, about 0.1 MPa to 45 MPa, about 0.1 MPa to 40 MPa, about 0.1 MPa to 35 MPa, about 0.1 MPa to 30 MPa, or about 0.1 MPa to 25 MPa. In certain embodiments, the pressure of the isochoric freezing is in the range of about 0.1 MPa to 35 MPa or about 0.1 MPa to 60 MPa. In some embodiments, the pressure of the isochoric freezing is about 150 MPa, about 145 MPa, about 140 MPa, about 135 MPa, about 130 MPa, about 125 MPa, about 120 MPa, about 115 MPa, about 110 MPa, about 105 MPa, about 100 MPa, about 95 MPa, about 90 MPa, about 85 MPa, about 80 MPa, about 75 MPa, about 70 MPa, about 65 MPa, about 60 MPa, about 55 MPa, about 50 MPa, about 45 MPa, about 40 MPa, about 35 MPa, about 30 MPa, about 25 MPa. In other embodiments, the pressure of the isochoric freezing is about 150 MPa or less. In other embodiments, the pressure of the isochoric freezing is about 100 MPa or less. In certain embodiments, the pressure of the isochoric freezing is about 35 MPa, about 30 MPa, about 25 MPa, about 20 MPa, about 15 MPa, or about 12 MPa.

[0132] In some embodiments, the isochoric freezing is at about -2 °C, under about 26 MPa for about 96 hours. In some embodiments, the isochoric freezing is at about -6 °C, under about 65 MPa for about 48 hours. In some embodiments, the isochoric freezing is at about -7 °C, under about 75 MPa for about 72 hours.

[0133] In some embodiments, the isochoric freezing conditions combining the abovedescribed subfreezing temperature and the pressure which occurs at the above-described subfreezing temperature affect connective tissues of shellfish and reduce or inhibit enzymatic activity of shellfish, thereby opening shells of the shellfish and detaching a meat inside of the shellfish. The isochoric freezing reduces or inhibits enzymatic activity in the shellfish and weakens adhesion of the shellfish to its shells. b-2) isochoric freezing for partially opening shells of shellfishAttorney Docket: BCHR-002WO

[0134] In some embodiments, the method for partially opening shells of shellfish comprises b-2) isochoric freezing the fluid in the isochoric chamber with a subfreezing temperature of -0.1 °C or less, a pressure of 1 MPa or more, and a treatment time of 30 minutes.

[0135] In some embodiments, the treatment time for isochoric freezing is 30 minutes or more. In some embodiments, the treatment time for isochoric freezing is 1 hour or more. In some embodiments, the treatment time for isochoric freezing is 6 hours or more. In some embodiments, the treatment time for isochoric freezing is 12 hours or more. In some embodiments, the treatment time for isochoric freezing is 24 hours or more. In some embodiments, the treatment time for isochoric freezing is 36 hours or more. In some embodiments, the treatment time for isochoric freezing is 48 hours or more. In some embodiments, the treatment time for isochoric freezing is 72 hours or more. In certain embodiments, the treatment time for isochoric freezing is about 30 minutes to 96 hours. In certain embodiments, the treatment time for isochoric freezing is about 1 hour to 96 hours. In certain embodiments, the treatment time for isochoric freezing is about 6 hours to 96 hours. In certain embodiments, the treatment time for isochoric freezing is about 12 hours to 72 hours. In certain embodiments, the treatment time for isochoric freezing is about 24 hours to 72 hours. In certain embodiments, the treatment time for isochoric freezing is about 36 hours to 72 hours. In certain embodiments, the treatment time for isochoric freezing is about 48 hours to 72 hours. For example, the treatment time for isochoric freezing is about 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 75 hours, 78 hours, 80 hours, 83 hours, 85 hours, 87 hours, 90 hours, 93 hours, or 96 hours. In certain embodiments, the treatment time for isochoric freezing is about 72 hours.Attorney Docket: BCHR-002WO

[0136] In some embodiments, the subfreezing temperature is -0.1 °C or less. In some embodiments, the subfreezing temperature is -0.5 °C or less, -1.0 °C or less, -1.5 or less, -2.0 °C or less, -2.5 °C or less, or -3.0 °C or less. In some embodiments, the subfreezing temperature is in the range of about -0.1 °C to -3.5 °C. In some embodiments, the subfreezing temperature is in the range of about -1.0 °C to -3 °C. In some embodiments, the subfreezing temperature is in the range of about -2.0 °C to -3 °C. In some embodiments, the subfreezing temperature is in the range of about -2.5 °C to -3 °C. In some embodiments, the subfreezing temperature is about -0.1 °C, -0.2 °C, -0.3 °C, -0.4 °C, -0.5 °C, -0.6 °C, -0.7 °C, -0.8 °C, -0.9 °C, -1.0 °C, -1.1 °C, -1.2 °C, -1.3 °C, -1.4 °C, -1.5 °C, -1.6 °C, -1.7 °C, -1.8 °C, -1.9 °C, -2.0 °C, -2.1 °C, -2.2 i, -2.3 °C, -2.4 °C, -2.5 °C, -2.6 °C, -2.7 °C, -2.8 °C, -2.9 °C, -3.0 °C, -3.1 °C, -3.2 °C, -3.3 °C, -3.4 °C, or -3.5 °C.In certain embodiments, the subfreezing temperature of the present disclosure is about -3.0 °C.

[0137] In some embodiments, the isochoric freezing is at a pressure of 1 MPa or more. In some embodiments, the isochoric freezing is at a pressure of 5 MPa or more. In some embodiments, the isochoric freezing is at a pressure of 10 MPa or more. In some embodiments, the isochoric freezing is at a pressure of 15 MPa or more. In some embodiments, the isochoric freezing is at a pressure of 20 MPa or more. In some embodiments, the isochoric freezing is at a pressure of 25 MPa or more. In some embodiments, the isochoric freezing is at a pressure of 30 MPa or less. In some embodiments, the isochoric freezing is at a pressure of 35 MPa or less. In some embodiments, the isochoric freezing is at a pressure in the range of about 1 MPa to 35 MPa. In some embodiments, the isochoric freezing is at a pressure in the range of about 1 MPa to 30 MPa. In some embodiments, the isochoric freezing is at a pressure in the range of about 5 MPa to 30 MPa. In some embodiments, the isochoric freezing is at a pressure in the range of about 10 MPa to 30 MPa. In some embodiments, the isochoric freezing is at a pressure in the range of about 15 MPa to 30 MPa. In some embodiments, the isochoric freezing is at a pressure in the range of about 20 MPa to 30 MPa. In some embodiments, the isochoric freezing is at a pressure in the range of about 25 MPa to 30 MPa. For example, the isochoric freezing is at a pressure of about 1 MPa, 2 MPa, 3 MPa, 4 MPa, 5 MPa, 6 MPa, 7 MPa, 8 MPa, 9 MPa, 10 MPa, 11 MPa, 12 MPa, 13 MPa, 14 MPa, 15 MPa, 16 MPa, 17 MPa, 18 MPa, 19 MPa, 20 MPa, 21 MPa, 22 MPa, 23Attorney Docket: BCHR-002WOMPa, 24 MPa, 25 MPa, 26 MPa, 27 MPa, 28 MPa, 29 MPa, 30 MPa, 31 MPa, 32 MPa, 33 MPa, 34 MPa, or 35 MPa. In certain embodiments, the isochoric freezing is at a pressure of about 30 MPa.

[0138] In some embodiments, the isochoric freezing condition for partially opening shells of shellfish is a subfreezing temperature of -0.1 °C or less, a pressure of 1 MPa or more, and a treatment time of 30 minutes or more. In certain embodiments, the isochoric freezing condition for partially opening the shellfish is a subfreezing temperature of about -0.1 °C to - 3.5 °C, a pressure of about 1 MPa to 35 MPa and a treatment time of about 30 minutes to 72 hours. In certain embodiments, the isochoric freezing condition for partially opening the shellfish is a subfreezing temperature of about -0.1 °C to -3.0 °C, a pressure of about 1 MPa to 30 MPa and a treatment time of about 30 minutes to 48 hours. In certain embodiments, the isochoric freezing condition for partially opening the shellfish is a subfreezing temperature of about -1.0 °C to -3.0 °C, a pressure of about 10 MPa to 30 MPa and a treatment time of about 12 hours to 72 hours. In certain embodiments, the isochoric freezing condition for partially opening the shellfish is a subfreezing temperature of about -2.0 °C to -3.0 °C, a pressure of about 20 MPa to 30 MPa and a treatment time of about 36 hours to 72 hours. In certain embodiments, the isochoric freezing condition for partially opening the shellfish is a subfreezing temperature of about -2.5 °C to -3.0 °C, a pressure of about 25 MPa to 30 MPa and a treatment time of about 48 hours to 72 hours. In certain embodiments, the isochoric freezing condition for partially opening the shellfish is a subfreezing temperature of about -3.0 °C, a pressure of about 30 MPa and a treatment time of about 72 hours.

[0139] In some embodiments, through the isochoric freezing treatment method of the present disclosure, the shellfish remains mostly closed but is structurally weakened, thereby retaining liquor and avoiding meat dehydration. The attachment of the meat to the shells is also weakened. Herein, partially opening shellfish includes partially detaching the meat from the shells of the shellfish. c) Warming the isochoric chamber containing the shellfish

[0140] In some embodiments, the method further comprises c) warming the fluid in the isochoric chamber containing the shellfish. Once the shellfish is incubated under isochoric freezing over the treatment time, the chamber is gradually warmed above the freezingAttorney Docket: BCHR-002WO temperature of the water, allowing the ice in the chamber to melt and the pressure to decrease. The chamber is then opened, and the shellfish is taken out from the chamber. After the isochoric freezing treatment, the shellfish is opened and meat inside of the shellfish is detached. The thawing may be achieved by ceasing active cooling and allowing simple natural convection with surrounding air at a temperature above 0°C; by forced convection from a warm (equal to or above 0°C) fluid source; by forced convection from a warming blanket wrapped around the chamber, through which a warm fluid is circulated; by radiation from a heat lamp or other radiative heat source; by direct electromagnetic or joule heating; by conduction from a heating block that is itself warmed by any of the means above; or by any other means of active or passive heat transfer.

[0141] In some embodiments, the warming or thawing time is in the range of about 1 minute to 24 hours, such as, about 1 minute to 22 hours, about 1 minute to 20 hours, about 1 minute to 18 hours, about 1 minute to 15 hours, about 1 minute to 13 hours, about 1 minute to 12 hours, about 1 minute to 10 hours, about 1 minute to 8 hours, about 1 minute to 5 hours, about 5 minutes to 24 hours, about 10 minutes to 24 hours, about 15 minutes to 24 hours, about 20 minutes to 24 hours, about 30 minutes to 24 hours, about 30 minutes to 2 hours, about 30 minutes to 3 hours, about 30 minutes to 4 hours, about 30 minutes to 5 hours, about 30 minutes to 6 hours, about 30 minutes to 7 hours, about 30 minutes to 8 hours, about 30 minutes to 9 hours, about 30 minutes to 10 hours, about 30 minutes to 11 hours, about 30 minutes to 12 hours, about 30 minutes to 15 hours, about 30 minutes to 18 hours, about 30 minutes to 20 hours, or about 30 minutes to 22 hours.

[0142] In some embodiments, the thawing or warming temperature is above 0 °C to 5 °C. For example, the thawing or warming temperature is above 0 °C to 5 °C, above 0 °C to 6 °C, above 0 °C to 7 °C, above 0 °C to 8 °C, above 0 °C to 9 °C, above 0 °C to 10 °C, above 0 °C to 12 °C, above 0 °C to 15 °C, above 0 °C to 20 °CJabove 0 °C to 22 °C, above 0 °C to 25 °C, above 0 °C to 27 °C, above 0 °C to 30 °C, above 0 °C to 35 °C, above 0 °C to 40 °C, above 0 °C to 45 °C, above 0 °C to 50 °C, above 0 °C to 55 °C, above 0 °C to 60 °C, above 0 °C to 65 °C, above 0 °C to 70 °C, above 0 °C to 75 °C, above 0 °C to 80 °C, above 0 °C to 85 °C above 0 °C to 90 °C, above 0 °C to 95 °C or above 0 °C and 100 °C or less.Attorney Docket: BCHR-002WO

[0143] In summary, the combined effects of subfreezing temperature and mild pressure disrupt the connective tissue between shell and the meat in shellfish, weaken enzymatic activities, and cause phase transitions in lipids. This all contributes to the efficient release of shellfish from its shell, without denaturation of proteins.

[0144] In some aspects of the method, isochoric freezing can open shells and release meat from shells of shellfish without or with minimal quality loss during short term treatment. In some embodiments, the quality loss is determined by measuring color change, texture change, water content change, humidity loss, pH change, mass change, total soluble solids change, titratable acidity (TA) change, ascorbic acid (AA) change, or antioxidant activity. In addition, the shellfish can be partially opened, and the shellfish meat can be partially detached from the shells of the shellfish under the mild isochoric freezing conditions, as described in the present disclosure. The shellfish can remain mostly closed but is structurally weakened, thereby retaining liquor and avoiding meat dehydration. The attachment of the meat to the shells is also weakened.4. Devices for processing shellfish

[0145] In some aspects, the present disclosure provides a device for processing shellfish comprising a) an isochoric chamber filled with a fluid, wherein the isochoric chamber contains shellfish; b) a temperature control system configured to cool the fluid in the isochoric chamber containing shellfish to a subfreezing temperature over a treatment time and thaw or warm the fluid in the isochoric chamber containing the shellfish, wherein a meat inside the shellfish is detached through the isochoric freezing. Also, the shellfish is opened and a meat inside the shellfish is detached thorough the isochoric freezing.Isochoric freezing

[0146] Herein, the isochoric freezing refers to the freezing of aqueous solutions in a constant volume chamber. The term "constant volume chamber" is used interchangeably herein with "isochoric chamber" or "rigid chamber". In certain embodiments, the isochoric chamber can be made of grade 7075 aluminum and pressure-rated for up to 110 MPa. In certain embodiments, the chamber can be connected to an electronic pressure transducer toAttorney Docket: BCHR-002WO monitor the pressure inside the chamber. The isochoric freezing technique is described in US2007 / 0042337A1 and PCT / US24 / 33896, which are incorporated by reference herein.

[0147] In the isochoric freezing system of the present disclosure, one or more shellfish are placed inside an isochoric chamber filled with an aqueous solution. In the isochoric freezing, a substantial portion of the volume remains unfrozen which is associated with an increase in pressure. The process of isochoric freezing occurs along the liquidus line in a temperature-pressure phase diagram as the temperature is dropped because the volume in an isochoric chamber is constant.

[0148] The Isochoric freezing is described in "3. Methods for processing shellfish" and is incorporated into this section. a) An isochoric chamber filled with a fluid

[0149] In some embodiments, the device for processing shellfish comprises a) an isochoric chamber filled with a fluid, wherein the isochoric chamber contains shellfish.

[0150] In some embodiments, the number of shellfish for processing is at least 1, at least 10, at least 100, at least 1,000, at least 10,000, at least 100,000, at least 1,000,000, or at least 10,000,000.

[0151] In some embodiments, shellfish has one shell. In other embodiments, shellfish has two or more shells. In certain embodiments, shellfish having one shell may not need to be open for processing, such as abalone. In certain embodiments, shellfish having two or more shells may need to be open for processing, such as mussels.

[0152] In some embodiments, the shellfish can be different kinds of shellfish. In other embodiments, the shellfish can be the same kind of shellfish.

[0153] In some embodiments, the isochoric chamber filled with a fluid may contain shellfish having one shell. In other embodiments, the isochoric chamber filled with a fluid may contain shellfish having two or more shells.

[0154] In some embodiments, the isochoric chamber filled with a fluid may contain different kinds of shellfish. In other embodiments, the isochoric chamber filled with a fluid may contain the same kind of shellfish.

[0155] In some embodiments, the shellfish of the present disclosure refers to a shelled mollusk, such as a clam, a snail, an oyster as well as some less familiar animals, like a tusk shellAttorney Docket: BCHR-002WO and a chiton. In some embodiments, mollusks used as a food source by humans include, but not limited to, many species of clams, mussels, oysters, winkles, and scallops. More specifically, the shellfish of the present disclosure is selected from a bivalve, a univalve, a crustacean, a gastropod, a scaphopod, a polyplacophoran, or an echinoderm. In some embodiments, the shellfish of the present disclosure is edible. As such, the present disclosure applies to all types and species of shellfish.

[0156] In certain embodiments, the bivalve is selected from a mussel, a scallop, a clam, a winkle, and an oyster. For example, a mussel is, but not limited to, a blue mussel.

[0157] In certain embodiments, the univalve is a snail. For example, univalve includes, but is not limited to, a burgundy snail, a giant African snail, a cerithidea obtusa, an apple snail, a red-rimmed melania, a cone snail, a helix, a patella vulgate, a milk snail.

[0158] In certain embodiments, the crustacean is selected from a shrimp, a prawn, a crab, a lobster, barnacles, and crayfish. Also, the crustacean is an isopod, a decapod, or a woodlouse. For example, the shrimp is, but not limited to, a caridean shrimp, a mantis shrimp, a brine shrimp, a whiteleg shrimp, a prawn, and a krill. For example, the lobster is, but not limited to, a spiny lobster, slipper lobster. For example, the crab is, but is not limited to, a coconut crab, a king crab, a red king crab, a mole crab, a true crab or a hermit crab.

[0159] In certain embodiments, the gastropod is, but not limited to, a conch, an abalone, a limpet, and a whelk.

[0160] In certain embodiments, the scaphopoda includes, but is not limited to, a dentalium, a fissidentalium, a gadilida, a gadila, and a gadilida.

[0161] In certain embodiments, echinoderm includes, but is not limited to, starfish, brittle stars, sea urchins, sand dollars and sea cucumbers, as well as the sessile sea lilies.

[0162] In some embodiments, the shellfish is alive prior to the isochoric freezing. In other embodiments, the shellfish is raw shellfish or undercooked shellfish.

[0163] In some embodiments, the device of the present disclosure is suitable for bulk processing. In some embodiments, the volume of the isochoric chamber is suitable for bulk processing. In certain embodiments, the volume of the isochoric chamber is about 50 L, about 100 L, about 1,000 L, about 1,500 L, about 2,000 L, about 3,000 L, about 4,000 L, about 5,000 L, or about 10,000 L. In certain embodiments, the volume of the isochoric chamber is about 50 LAttorney Docket: BCHR-002WO to 10,000 L, about 100 L to 10,000 L, about 500 L to 10,000 L, about 1,000 L to 10,000 L, 1,000 L to 9,000 L, 1,000 L to 8,000 L, 1,000 L to 7,000 L, 1,000 L to 6,000 L, 1,000 L to 5,000 L, 1,000 L to 4,000 L, 1,000 L to 3,000 L, or 1,000 L to 2,000 L. . In certain embodiments, the volume of the isochoric chamber is about 1,000 L to 2,000 L.

[0164] For bulk processing, the isochoric chamber contains a large number of shellfish. For example, the large number of shellfish is at least 50, at least 80, at least 100, at least 500, at least 1,000, at least 1,500, at least 2,000, at least 3,000, at least 4,000, at least 5,000, at least 6,000, at least 7,000, at least 8,000, at least 9,000, at least 10,000, at least 50,000, at least 100,000, at least 500,000, or at least 1,000,000.

[0165] In some embodiments, meat of the large number of shellfish are detached from the shellfish through the isochoric freezing. In other embodiments, the large number of shellfish are opened and meat inside the shellfish are detached through the isochoric freezing. In certain embodiments, 30 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In other embodiments, 40 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In still other embodiments, 50 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In yet other embodiments, 60 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In further embodiments, 70 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In still other embodiments, 80 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In yet other embodiments, 90 % or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, where the remaining shellfish are closed or partially opened. In further embodiments, all of the shellfish are open and the meat inside the shellfish are detached through the isochoric freezing.Attorney Docket: BCHR-002WO

[0166] In some embodiments, the shellfish can be directly placed in the isochoric chamber. FIG. 4 illustrates an exemplary embodiment of isochoric freezing configuration with an isochoric chamber for a biological mass while avoiding the formation of ice. In other embodiments, a biological matter can be placed in a container that allows transfer of pressure mass and heat. In still other embodiments, a biological matter can be placed in a container that allows transfer of only pressure and heat but not mass.

[0167] In other embodiments, the shellfish is placed in a matter container and the matter container is placed in the fluid in the isochoric chamber. FIG. 5 illustrates an exemplary embodiment of isochoric freezing configuration with an isochoric chamber and a matter container for including a biological mass while avoiding the formation of ice. Also, FIG. 5 illustrates an exemplary embodiment of isochoric freezing configuration with an isochoric chamber and a matter container for including a biological material while avoiding the formation of ice. The matter container transfer pressure but not mass in the isochoric chamber. In some embodiments, the matter container includes a biological material. In other embodiments, the matter container including a biological material is filled with a fluid that does not freeze at the isochoric temperature. Herein, the biological material can be shellfish.

[0168] In some embodiments, shellfish is vacuum packed and placed in the isochoric chamber. For example, shellfish is collected in a polyethylene sterile vacuum bag and vacuum is then created within the bag and placed inside the isochoric chamber. In other embodiments, shellfish is collected in a polyethylene sterile bag. The bag is then sealed with negligible headspace and placed inside the isochoric chamber. In other embodiments, shellfish is not vacuum packed and is placed directly in a fluid in an isochoric chamber.

[0169] In some embodiments, the fluid in the constant volume chamber is aqueous solution or pure water with or without organic molecules therein. In some embodiments, the aqueous solution that is primarily water may contain other chemicals so that the freezing point of the water-based solution may be modified as required for a specific application. For example, the water-based solution may contain salt. In certain embodiments, the external solution is water. In some embodiments, the fluid in the container can be the same as the fluid in the isochoric chamber surrounding the container. In other embodiments, the fluid in the matter container can be different from the fluid in the isochoric chamber surrounding theAttorney Docket: BCHR-002WO matter container. In other words, the fluid inside the matter container and the fluid outside the matter container can be of different types in osmotic equilibrium with the preserved biological matter. In some embodiments the isochoric chamber is filled with a fluid that does not freeze at the storage temperature, such as a solution of glycerol.

[0170] In some embodiments, the isochoric chamber may contain a nucleating agent. The nucleating agent forms an ice crystal thereby the shellfish can be stored in the unfrozen part of the volume. The ice crystal formed by the nucleating agent generates pressures of the isochoric chamber, and the enhanced pressures hinder further formation of ice. In some embodiments, the nucleating agent can be a structural element of the isochoric freezing device. For example, the nucleating agent can be walls of the isochoric chamber itself, or walls of the matter container itself. In other embodiments, the nucleating agent can be any agent that promotes the formation of ice crystal in the isochoric chamber. For example, the ice nucleating agent is, but not limited to, minute solid particles, such as dust or food particles, large molecules, ice-nucleating proteins. In some embodiments, the nucleating agent is placed in the bottom of the isochoric chamber, and the chamber is completely filled with an aqueous solution. In some embodiments, shellfish is placed in a container in an isochoric chamber that allows transfer of only pressure and heat but not mass and the isochoric chamber is filled with a fluid and nucleation agents that cause the fluid to freeze before the shellfish in the container freezes. b) A temperature control system of the device

[0171] The device of the present disclosure also comprises b) a temperature control system configured to cool the fluid in the isochoric chamber containing the shellfish to a subfreezing temperature over a treatment time and warm the fluid in the isochoric chamber containing the shellfish. The Isochoric freezing is described in "3. Methods for processing shellfish" and is incorporated into this section.

[0172] In some embodiments, the temperature control system is configured to cool the fluid in the isochoric chamber containing the shellfish to a subfreezing temperature over a treatment time and thaw or warm the fluid in the isochoric chamber containing the shellfish above the freezing temperature of the water. In some embodiments, the temperature control system can be any device configured to cool and warm the fluid in the isochoric chamberAttorney Docket: BCHR-002WO containing the shellfish. For example, the temperature control system is, but not limited to, a thermostatic control system, a proportional-integral-derivative controller, programmable logic controllers, simple relays or on / off switches, or other means of implementing active or passive control logic in a thermal system.

[0173] In some embodiments, the temperature control system comprises a cooling system and a heating system configured to cool and warm the fluid in the isochoric chamber containing the shellfish. In certain embodiments, the cooling system can be any device configured to cool the fluid in the isochoric chamber containing the shellfish, such as a conventional freezer or other types of temperature cabinets or boxes, cooling blankets that enshroud the chamber and have chilled fluids circulating through them, liquid cooling baths with fluids that stay liquid at sub-zero temperatures, and any other system by which heat may be extracted from the chamber and / or fluid within. In certain embodiments, the fluid in the isochoric chamber can be cooled by dry ice. In certain embodiments, the heating system is a conventional heater, an electronic blanket, a blanket with a circulating warm fluid within, or any device configured to warm the isochoric chamber. In exemplary embodiments, the isochoric chamber may be surrounded by an electronic blanket to be warm above the freezing temperature of the water. In exemplary embodiments, the isochoric chamber may be placed into a bigger container containing warm water. In exemplary embodiments, the isochoric chamber may be surrounded by a blanket with a fluid circulating through it, and said fluid may be circulated by an external bath capable of cooling or warming the fluid. In this embodiment, both cooling and warming may be achieved by the same blanket, by changing the temperature of the bath that is circulating the fluid.

[0174] In some embodiments of the device, the treatment time refers to the time applied for isochoric freezing. In other embodiments, the time applied for isochoric freezing is the time for treating shellfish product in an isochoric chamber. In some embodiments, the treatment time of the present disclosure refers to a length of time from minutes to hours. In other embodiments, the treatment time of the present disclosure refers to a length of time from hours to days. In some embodiments, the treatment time for isochoric freezing is not limited as long as applying isochoric freezing conditions to a shellfish product in an isochoric chamber.Attorney Docket: BCHR-002WO

[0175] In certain embodiments, the treatment time for isochoric freezing is, but not limited to, 10, 15, 20, 25, 30, 40, or 50 days. In certain embodiments, the treatment time for isochoric freezing is 1 hour or more, 4 hours or more, 12 hours or more, 16 hours or more, 20 hours or more, or 24 hours or more. In some embodiments, the treatment time is, but not limited to about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 75 hours, 78 hours, 80 hours, 83 hours, 85 hours, 87 hours, 90 hours, 93 hours, or 96 hours. In certain embodiments, the treatment time for isochoric freezing is 3 hours or less. In certain embodiments, the treatment time for isochoric freezing is 4 hours or less. In certain embodiments, the treatment time for isochoric freezing is 5 hours or less. In certain embodiments, the treatment time for isochoric freezing is 6 hours or less.

[0176] In some embodiments, the treatment time for isochoric freezing is in the range of about 1 hour to 50 days. In certain embodiments, the treatment time for isochoric freezing is in the range of about 1 hour to 2 hours, about 1 hour to 3 hours, about 1 hour to 4 hours, about 1 hour to 5 hours, about 1 hour to 6 hours, about 1 hour to 7 hours, about 1 hour to 8 hours, about 1 hour to 9 hours, about 1 hour to 10 hours, about 1 hour to 11 hours, about 1 hour to 12 hours, about 1 hour to 13 hours, about 1 hour to 14 hours, about 1 hour to 15 hours, about 1 hour to 16 hours, about 1 hour to 17 hours, about 1 hour to 18 hours, about 1 hour to 19 hours, about 1 hour to 20 hours, about 1 hour to 21 hours, about 1 hour to 22 hours, about 1 hour to 23 hours, about 1 hour to 24 hours, 1 hour to 48 hours, 1 hour to 72 hours, or 1 hour to 96 hours. In certain embodiments, the treatment time for isochoric freezing is about 48 hours. In certain embodiments, the treatment time for isochoric freezing is about 72 hours. In certain embodiments, the treatment time for isochoric freezing is about 96 hours.Attorney Docket: BCHR-002WO

[0177] In some aspects of the device, the subfreezing temperature for processing shellfish is much higher than the triple point along the liquidus line in a temperature pressure phase diagram depicted in FIG. 1. The triple point in the temperature pressure phase diagram depicted in FIG. 1 is about -21 °C. In some embodiments, the subfreezing temperature of the present disclosure is at least 5 °C higher than the triple point along the liquidus line in a temperature pressure phase diagram depicted in FIG. 1. In other embodiments, the subfreezing temperature of the present disclosure is at least 10 °C higher than the triple point along the liquidus line in a temperature pressure phase diagram depicted in FIG. 1. In some embodiments, the subfreezing temperature of the present disclosure is in the range of about 0 °C to -20 °C, about 0 °C to -15 °C, about 0 °C to -10 °C, about 0 °C to -5 °C, or about 0 °C to -2 °C. In some embodiments, the subfreezing temperature of the present disclosure is in the range of about 0 °C to -20 °C, 0 °C to -15 °C, 0 °C to -14 °C, 0 °C to -13 °C, 0 °C to -12 °C, 0 °C to -11 °C, 0 °C to -10 °C, 0 °C to -9 °C, 0 °C to -8 °C, 0 °C to -7 °C, 0 °C to -6 °C, 0 °C to -5 °C, or 0 °C to -2 °C. In some embodiments, the subfreezing temperature of the present disclosure is about -1 °C, -2 °C, -3 °C, -4 °C, -5 °C, -6 °C, -7 °C, -8 °C, -9 °C, -10 °C, -11 °C, -12 °C, -13 °C, or -14 °C.

[0178] In some embodiments of the device, the isochoric freezing conditions combining the above-described subfreezing temperature and the pressure which occurs at the above-described subfreezing temperature when the closed isochoric chamber is in a state of thermodynamic equilibrium is effective in opening shells of shellfish and releasing the meat inside. Furthermore, the pressures induced by the isochoric freezing of the present disclosure have further effects on avoiding ice formation inside the shellfish at the subfreezing temperature. In the device of the present disclosure, the applied pressure occurs at the abovedescribed subfreezing temperature when the closed isochoric chamber is in a state of thermodynamic equilibrium. Therefore, the isochoric system of the present disclosure provides convenient ways to achieve subfreezing temperature and elevated pressures only by controlling the temperature without the need for mechanical means to elevated pressure.

[0179] In some embodiments, the isochoric freezing of the present disclosure is at a pressure in the range of about 0.1 MPa to 150 MPa, about 0.1 MPa to 145 MPa, about 0.1 MPa to 140 MPa, about 0.1 MPa to 135 MPa, about 0.1 MPa to 130 MPa, about 0.1 MPa to 125 MPa, about 0.1 MPa to 120 MPa, about 0.1 MPa to 115 MPa, about 0.1 MPa to 110 MPa,Attorney Docket: BCHR-002WO about 0.1 MPa to 105 MPa, about 0.1 MPa to 100 MPa, about 0.1 MPa to 95 MPa, about 0.1 MPa to 90 MPa, about 0.1 MPa to 85 MPa, about 0.1 MPa to 80 MPa, about 0.1 MPa to 75 MPa, about 0.1 MPa to 70 MPa, about 0.1 MPa to 65 MPa, about 0.1 MPa to 60 MPa, about 0.1 MPa to 55 MPa, about 0.1 MPa to 50 MPa, about 0.1 MPa to 45 MPa, about 0.1 MPa to 40 MPa, about 0.1 MPa to 35 MPa, about 0.1 MPa to 30 MPa, or about 0.1 MPa to 25 MPa. In certain embodiments, the pressure of the isochoric freezing is in the range of about 0.1 MPa to 35 MPa or about 0.1 MPa to 60 MPa. In some embodiments, the pressure of the isochoric freezing is about 150 MPa, about 145 MPa, about 140 MPa, about 135 MPa, about 130 MPa, about 125 MPa, about 120 MPa, about 115 MPa, about 110 MPa, about 105 MPa, about 100 MPa, about 95 MPa, about 90 MPa, about 85 MPa, about 80 MPa, about 75 MPa, about 70 MPa, about 65 MPa, about 60 MPa, about 55 MPa, about 50 MPa, about 45 MPa, about 40 MPa, about 35 MPa, about 30 MPa, or about 25 MPa. In other embodiments, the pressure of the isochoric freezing is about 150 MPa or less. In other embodiments, the pressure of the isochoric freezing is about 100 MPa or less. In certain embodiments, the pressure of the isochoric freezing is about 35 MPa, about 30 MPa, about 25 MPa, about 20 MPa, about 15 MPa, or about 12 MPa.

[0180] In some embodiments, the isochoric freezing is at about -2 °C, under about 26 MPa for about 96 hours. In some embodiments, the isochoric freezing is at about -6 °C, under about 65 MPa for about 48 hours. In some embodiments, the isochoric freezing is at about -7 °C, under about 75 MPa for about 72 hours.

[0181] In some embodiments, the isochoric freezing conditions combining the abovedescribed subfreezing temperature and the pressure which occurs at the above-described subfreezing temperature affect connective tissues of shellfish and reduce or inhibit enzymatic activity of shellfish, thereby opening shells of the shellfish and detaching a meat inside of the shellfish. The isochoric freezing reduces or inhibits enzymatic activity in the shellfish and weakens adhesion of the shellfish to its shells.

[0182] In some embodiments, the isochoric freezing condition for partially opening shells of shellfish is a subfreezing temperature of -0.1 °C or less, a pressure of 1 MPa or more, and a treatment time of 30 minutes or more.Attorney Docket: BCHR-002WO

[0183] In some embodiments, the treatment time for isochoric freezing is 30 minutes or more. In some embodiments, the treatment time for isochoric freezing is 1 hour or more. In some embodiments, the treatment time for partially opening shells of shellfish is 6 hours or more. In some embodiments, the treatment time for partially opening shells of shellfish is 12 hours or more. In some embodiments, the treatment time for partially opening shells of shellfish is 24 hours or more. In some embodiments, the treatment time for partially opening shells of shellfish is 36 hours or more. In some embodiments, the treatment time for partially opening shells of shellfish is 48 hours or more. In some embodiments, the treatment time for partially opening shells of shellfish is 72 hours or more. In certain embodiments, the treatment time for partially opening shells of shellfish is about 30 minutes to 96 hours. In certain embodiments, the treatment time for partially opening shells of shellfish is about 1 hour to 96 hours. In certain embodiments, the treatment time for partially opening shells of shellfish is about 6 hours to 96 hours. In certain embodiments, the treatment time for partially opening shells of shellfish is about 12 hours to 72 hours. In certain embodiments, the treatment time for partially opening shells of shellfish is about 24 hours to 72 hours. In certain embodiments, the treatment time for partially opening shells of shellfish is about 36 hours to 72 hours. In certain embodiments, the treatment time for partially opening shells of shellfish is about 48 hours to 72 hours. For example, the treatment time for partially opening shells of shellfish is about 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 75 hours, 78 hours, 80 hours, 83 hours, 85 hours, 87 hours, 90 hours, 93 hours, or 96 hours. In certain embodiments, the treatment time for partially opening shells of shellfish is about 72 hours.Attorney Docket: BCHR-002WO

[0184] In some embodiments, the subfreezing temperature for partially opening shells of shellfish is -0.1 °C or less. In some embodiments, the subfreezing temperature for partially opening shells of shellfish is -0.5 °C or less, -1.0 °C or less, -1.5 or less, -2.0 °C or less, -2.5 °C or less, or -3.0 °C or less. In some embodiments, the subfreezing temperature for partially opening shells of shellfish is in the range of about -0.1 °C to -3.5 °C. In some embodiments, the subfreezing temperature for partially opening shells of shellfish is in the range of about -1.0 °C to -3 °C. In some embodiments, the subfreezing temperature for partially opening shells of shellfish is in the range of about -2.0 °C to -3 °C. In some embodiments, the subfreezing temperature for partially opening shells of shellfish is in the range of about -2.5 °C and -3 °C. In some embodiments, the subfreezing temperature for partially opening shells of shellfish is about -0.1 °C, -0.2 °C, -0.3 °C, -0.4 °C, -0.5 °C, -0.6 °C, -0.7 °C, -0.8 °C, -0.9 °C, -1.0 °C, -1.1 °C, -2.4 °C, -2.5 °C, -2.6 °C, -2.7 °C, -2.8 °C, -2.9 °C, -3.0 °C, -3.1 °C, -3.2 °C, -3.3 °C, -3.4 °C, or -3.5 °C.In certain embodiments, the subfreezing temperature of for partially opening shells of shellfish is about -3.0 °C.

[0185] In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure of 1 MPa or more. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure of 5 MPa more. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure of 10 MPa or more. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure of 15 MPa or more. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure of 20 MPa or more. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure of 25 MPa or more. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure of 30 MPa or less. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure of 35 MPa or less. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure in the range of about 1 MPa to 35 MPa. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure in the range of about 1 MPa to 30 MPa. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure in the range of about 5 MPa to 30 MPa.Attorney Docket: BCHR-002WOIn some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure in the range of about 10 MPa to 30 MPa. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure in the range of about 15 MPa to 30 MPa. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure in the range of about 20 MPa to 30 MPa. In some embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure in the range of about 25 MPa to 30 MPa. For example, the isochoric freezing for partially opening shells of shellfish is at a pressure of about 1 MPa, 2 MPa, 3 MPa, 4 MPa, 5 MPa, 6 MPa, 7 MPa, 8 MPa, 9 MPa, 10 MPa,11 MPa, 12 MPa, 13 MPa, 14 MPa, 15 MPa, 16 MPa, 17 MPa, 18 MPa, 19 MPa, 20 MPa, 21 MPa, 22 MPa, 23 MPa, 24 MPa, 25 MPa, 26 MPa, 27 MPa, 28 MPa, 29 MPa, 30 MPa, 31 MPa, 32 MPa, 33 MPa, 34 MPa, or 35 MPa. In certain embodiments, the isochoric freezing for partially opening shells of shellfish is at a pressure of about 30 MPa.

[0186] In some embodiments, the isochoric freezing condition for partially opening shells of shellfish is a subfreezing temperature of -0.1 °C or less, a pressure of 1 MPa or more, and a treatment time of 30 minutes or more. In certain embodiments, the isochoric freezing condition for partially opening the shellfish is a subfreezing temperature of about -0.1 °C to - 3.5 °C, a pressure of about 1 MPa to 35 MPa and a treatment time of about 30 minutes to 72 hours. In certain embodiments, the isochoric freezing condition for partially opening the shellfish is a subfreezing temperature of about -0.1 °C to -3.0 °C, a pressure of about 1 MPa to 30 MPa and a treatment time of about 30 minutes to 48 hours. In certain embodiments, the isochoric freezing condition for partially opening the shellfish is a subfreezing temperature of about -1.0 °C to -3.0 °C, a pressure of about 10 MPa to 30 MPa and a treatment time of about12 hours to 72 hours. In certain embodiments, the isochoric freezing condition for partially opening the shellfish is a subfreezing temperature of about -2.0 °C to -3.0 °C, a pressure of about 20 MPa to 30 MPa and a treatment time of about 36 hours to 72 hours. In certain embodiments, the isochoric freezing condition for partially opening the shellfish is a subfreezing temperature of about -2.5 °C to -3.0 °C, a pressure of about 25 MPa to 30 MPa and a treatment time of about 48 hours to 72 hours. In certain embodiments, the isochoric freezing for partially opening shells of shellfish is a subfreezing temperature of about -3.0 °C, a pressure of about 30 MPa and a treatment time of about 72 hours.Attorney Docket: BCHR-002WO

[0187] In some embodiments, the warming or thawing time is in the range of about 1 minute to 24 hours, such as, about 1 minute to 22 hours, about 1 minute to 20 hours, about 1 minute to 18 hours, about 1 minute to 15 hours, about 1 minute to 13 hours, about 1 minute to 12 hours, about 1 minute to 10 hours, about 1 minute to 8 hours, about 1 minute to 5 hours, about 5 minutes to 24 hours, about 10 minutes to 24 hours, about 15 minutes to 24 hours, about 20 minutes to 24 hours, about 30 minutes to 24 hours, about 30 minutes to 2 hours, about 30 minutes to 3 hours, about 30 minutes to 4 hours, about 30 minutes to 5 hours, about 30 minutes to 6 hours, about 30 minutes to 7 hours, about 30 minutes to 8 hours, about 30 minutes to 9 hours, about 30 minutes to 10 hours, about 30 minutes to 11 hours, about 30 minutes to 12 hours, about 30 minutes to 15 hours, about 30 minutes to 18 hours, about 30 minutes to 20 hours, or about 30 minutes to 22 hours.

[0188] In some embodiments, the thawing or warming temperature is above 0 °C to 5 °C. For example, the thawing or warming temperature is above 0 °C to 5 °C, above 0 °C to 6 °C, above 0 °C to 7 °C, above 0 °C to 8 °C, above 0 °C to 9 °C, above 0 °C to 10 °C, above 0 °C to 12 °C, above 0 °C to 15 °C, above 0 °C to 20 °CJabove 0 °C to 22 °C, above 0 °C to 25 °CJabove 0 °C to 27 °C, above 0 °C to 30 °C, above 0 °C to 35 °C, above 0 °C to 40 °C, above 0 °C to 45 °C, above 0 °C to 50 °C, above 0 °C to 55 °C, above 0 °C to 60 °CJabove 0 °C to 65 °C, above 0 °C to 70 °Cjabove 0 °C to 75 °C, above 0 °C to 80 °C, above 0 °C to 85 °C above 0 °C to 90 °C, above 0 °C to 95 °C or above 0 °C and 100 °C or less. c) Optional elements

[0189] In some embodiments, the device of the present disclosure optionally includes a pressure monitoring element. In other embodiments, the device of the present disclosure optionally includes a pressure relief mechanism. In exemplary embodiments, the pressure monitoring element is a digital pressure transducer, analogue pressure gauge, or a strain gauge. In exemplary embodiments, the pressure relief mechanism is a pressure relief valve or a rupture disk.

[0190] In some embodiments, the device of the present disclosure optionally includes multiple discrete sub-systems for physically separating shellfish from ice. In exemplary embodiments, each of the sub-systems is a vacuum sealed polymer bag containing theAttorney Docket: BCHR-002WO shellfish. In exemplary embodiment, one or more of the sub-systems is a basket, mesh, or other container which localizes the shellfish towards the center of the chamber, and away from contact with the chamber walls, where ice will form initially.

[0191] In summary, the combined effects of subfreezing temperature and mild pressure disrupt the connective tissue between shell and the meat in shellfish, weaken enzymatic activities, and cause phase transitions in lipids. This all contributes to the efficient release of shellfish from its shell, without denaturation of proteins.

[0192] In some aspects of the device, the isochoric freezing can open shells and release the meat from the shells of the shellfish without or with minimal quality loss during short term treatment. In some embodiments, the quality loss is determined by measuring color change, texture change, water content change, humidity loss, pH change, mass change, total soluble solids change, titratable acidity (TA) change, ascorbic acid (AA) change, or antioxidant activity.

[0193] Certain aspects of the presently disclosed subject matter having been stated hereinabove, which are addressed in whole or in part by the presently disclosed subject matter, other aspects will become evident as the description proceeds when taken in connection with the accompanying Examples and Figures as best described herein below.EXAMPLES OF NON-LIMITING ASPECTS OF THE DISCLOSURE

[0194] Aspects, including embodiments, of the present subject matter described above may be beneficial alone or in combination, with one or more other aspects or embodiments. Without limiting the foregoing description, certain non-limiting aspects of the disclosure are provided below. As will be apparent to those of skill in the art upon reading this disclosure, each of the individually numbered aspects may be used or combined with any of the preceding or following individually numbered aspects. This is intended to provide support for all such combinations of aspects and is not limited to combinations of aspects explicitly provided below:

[0195] Aspect 1. A method for processing shellfish comprising: a) placing shellfish in an isochoric chamber filled with a fluid; andAttorney Docket: BCHR-002WO b) isochoric freezing the fluid in the isochoric chamber at a subfreezing temperature over a treatment time; wherein a meat inside the shellfish is detached through the isochoric freezing.

[0196] Aspect 2. The method of Aspect 1, further comprising c) warming the isochoric chamber containing the shellfish.

[0197] Aspect 3. The method of Aspect 1 or 2, wherein the treatment time is 1 hour to96 hours.

[0198] Aspect 4. The method of any one of Aspects 1-3, wherein the treatment time is1 hour to 48 hours.

[0199] Aspect 5. The method of Aspect 4, wherein the treatment time is from 1 hour to 24 hours.

[0200] Aspect 6. The method of any one of Aspects 1-5, wherein the subfreezing temperature is in the range of about 0 °C to -20 °C.

[0201] Aspect 7. The method of Aspect 6, wherein the subfreezing temperature is in the range of about 0 °C to -10 °C.

[0202] Aspect 8. The method of Aspect 6, wherein the subfreezing temperature is in the range of about 0 °C to -5 °C.

[0203] Aspect 9. The method of Aspect 6, wherein the subfreezing temperature is in the range of about 0 °C to -2 °C.

[0204] Aspect 10. The method of any one of Aspects 1 to 9, wherein the isochoric freezing is at a pressure in the range of about 0.1 MPa to about 150 MPa.

[0205] Aspect 11. The method of Aspect 10, wherein the isochoric freezing is at a pressure in the range of about 0.1 MPa to about 100 MPa.

[0206] Aspect 12. The method of Aspect 10, wherein the isochoric freezing is at a pressure in the range of about 0.1 MPa to about 55 MPa.

[0207] Aspect 13. The method of Aspect 10, wherein the isochoric freezing is at a pressure in the range of about 0.1 MPa to about 25 MPa.

[0208] Aspect 14. The method of any one of Aspects 1-13, wherein the method comprises placing a large number of shellfish in the isochoric chamber for bulk processing.Attorney Docket: BCHR-002WO

[0209] Aspect 15. The method of Aspect 14, wherein volume of the isochoric chamber is suitable for bulk processing.

[0210] Aspect 16. The method of Aspect 15, wherein the volume of the isochoric chamber is about 50L to 10,000 L.

[0211] Aspect 17. The method of Aspect 15, wherein the volume of the isochoric chamber is about 1,000 L to 2,000 L.

[0212] Aspect 18. The method of any one of Aspects 1-17, wherein the shellfish is placed directly in the isochoric chamber.

[0213] Aspect 19. The method of any one of Aspects 1 to 17, wherein the shellfish is placed in a matter container and the matter container is placed in the fluid in the isochoric chamber.

[0214] Aspect 20. The method of any one of Aspects 1-17, wherein the shellfish is vacuum packed and placed in the isochoric chamber.

[0215] Aspect 21. The method of any one of Aspects 1-20, wherein the isochoric chamber contains a nucleating agent.

[0216] Aspect 22. The method of any one of Aspects 1-21, wherein the fluid in the isochoric chamber is an aqueous solution or water.

[0217] Aspect 23. The method of any one of Aspects 1-22, wherein the shellfish is alive prior to the isochoric freezing.

[0218] Aspect 24. The method of any one of Aspects 1-23, wherein the shellfish is raw shellfish or undercooked shellfish.

[0219] Aspect 25. The method of any one of Aspects 1-24, wherein the shellfish is a bivalve, a univalve, a crustacean, a gastropod, a scaphopod, a polyplacophoran, or an echinoderm.

[0220] Aspect 26. The method of Aspect 25, wherein the bivalve is selected from a mussel, a scallop, a clam, a winkle, and an oyster.

[0221] Aspect 27. The method of Aspect 25, wherein the univalve is a snail.

[0222] Aspect 28. The method of Aspect 25, wherein the crustacean is selected from a shrimp, a prawn, a crab, a lobster, barnacles, and a crayfish.Attorney Docket: BCHR-002WO

[0223] Aspect 29. The method of Aspect 27, wherein the gastropod is selected from a conch, an abalone, a limpet, and a whelk.

[0224] Aspect 30. The method of any one of Aspects 1-29, wherein 30% or more, 50% or more, 70% or more, or 90% or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, wherein the remaining shellfish are closed or partially opened.

[0225] Aspect 31. The method of Aspect 30, wherein all of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing.

[0226] Aspect 32. The method of any one of Aspects 1-31, wherein the connective tissue of the shellfish to the shells is compromised by joint effects of subfreezing temperature and pressure under the conditions of isochoric freezing.

[0227] Aspect 33. The method of any one of Aspects 1-32, wherein the isochoric freezing reduces or inhibits enzymatic activity in the shellfish and weakens adhesion of the shellfish to its shells.

[0228] Aspect 34. A method for partially opening shellfish comprising: a) placing shellfish in an isochoric chamber filled with a fluid; and b-2) isochoric freezing the fluid in the isochoric chamber with a subfreezing temperature of -0.1 °C or less, a pressure of 1 MPa or more, and a treatment time of 30 minutes or more.

[0229] Aspect 35. The method of Aspect 34, comprising c) warming the isochoric chamber containing the shellfish.

[0230] Aspect 36. The method of Aspect 34 or 35, wherein the subfreezing temperature is about -0.1 °C to -3.0 °C, the pressure is 1 MPa to 35 MPa, and the treatment time is 48 hours or more.

[0231] Aspect 37. The method of Aspect 36, wherein the subfreezing temperature is about -2.5 °C to -3.0 °C, the pressure is about 25 MPa to 30 MPa and the treatment time is about 48 hours to 72 hours.

[0232] Aspect 38. The method of Aspect 36, wherein the subfreezing temperature is about -3.0 °C, the pressure is about 30 MPa and the treatment time is about 72 hours.Attorney Docket: BCHR-002WO

[0233] Aspect 39. The method of any one of Aspects 34-38, wherein the method comprises placing a large number of shellfish in the isochoric chamber for bulk processing.

[0234] Aspect 40. The method of any one of Aspects 34-39, wherein the shellfish is placed directly in the isochoric chamber.

[0235] Aspect 41. The method of any one of Aspects 34-39, wherein the shellfish is placed in a matter container, and the matter container is placed in the fluid in the isochoric chamber.

[0236] Aspect 42. The method of any one of Aspects 34-39, wherein the shellfish is vacuum packed and placed in the isochoric chamber.

[0237] Aspect 43. The method of any one of Aspects 34-42, wherein the isochoric chamber contains a nucleating agent.

[0238] Aspect 44. The method of any one of Aspects 34-43, wherein the fluid in the isochoric chamber is an aqueous solution or water.

[0239] Aspect 45. The method of any one of Aspects 34-44, wherein the shellfish is alive prior to the isochoric freezing.

[0240] Aspect 46. The method of any one of Aspects 34-44, wherein the shellfish is raw shellfish or undercooked shellfish.

[0241] Aspect 47. The method of any one of Aspects 34-46, wherein the shellfish is a bivalve, a univalve, a crustacean, a gastropod, a scaphopod, a polyplacophoran, or an echinoderm.

[0242] Aspect 48. The method of Aspect 47, wherein the bivalve is selected from a mussel, a scallop, a clam, a winkle, and an oyster.

[0243] Aspect 49. The method of Aspect 47, wherein the crustacean is selected from a shrimp, a prawn, a crab, a lobster, barnacles, and a crayfish.

[0244] Aspect 50. The method of Aspect 47, wherein the gastropod is selected from a conch, an abalone, a limpet, and a whelk.

[0245] Aspect 51. The method of Aspects 34-50, wherein the shellfish remains mostly closed but is structurally weakened, thereby retaining liquor and avoiding meat dehydration.

[0246] Aspect 52. A device for processing shellfish comprising:Attorney Docket: BCHR-002WO a) an isochoric chamber filled with a fluid, wherein the isochoric chamber contains shellfish; and b) a temperature control system configured to cool the fluid in the isochoric chamber containing the shellfish to a subfreezing temperature over a treatment time and warm the fluid in the isochoric chamber containing the shellfish.

[0247] Aspect 53. The device of Aspect 52, wherein the temperature control system comprises a cooling system and a heating system configured to cool and warm the fluid in the isochoric chamber containing the shellfish.

[0248] Aspect 54. The device of Aspect 52 or 53, wherein the device comprises a pressure monitoring element.

[0249] Aspect 55. The device of any one of Aspects 52-54, wherein the device comprises a pressure relief mechanism.

[0250] Aspect 56. The device of any one of Aspects 52-55, wherein the device comprises multiple discrete sub-systems for physically separating shellfish from ice.

[0251] Aspect 57. The device of any one of Aspects 52-56, wherein the treatment time is 1 hour to 96 hours.

[0252] Aspect 58. The device of Aspect 57, wherein the treatment time is 1 hour to 48 hours.

[0253] Aspect 59. The device of Aspect 57, wherein the treatment time is from 1 hour to 24 hours.

[0254] Aspect 60. The device of any one of Aspects 52-59, wherein the subfreezing temperature is in the range of about 0 °C to -20 °C.

[0255] Aspect 61. The device of Aspect 60, wherein the subfreezing temperature is in the range of about 0 °C to -10 °C.

[0256] Aspect 62. The device of Aspect 60, wherein the subfreezing temperature is in the range of about 0 °C to -5 °C.

[0257] Aspect 63. The device of Aspect 60, wherein the subfreezing temperature is in the range of about 0 °C to -2 °C.

[0258] Aspect 64. The device of any one of Aspects 51-63, wherein the isochoric freezing treatment is at a pressure in the range of about 0.1 MPa to about 150 MPa.Attorney Docket: BCHR-002WO

[0259] Aspect 65. The device of Aspect 64, wherein the isochoric freezing treatment is at a pressure in the range of about 0.1 MPa to about 100 MPa.

[0260] Aspect 66. The device of Aspect 64, wherein the isochoric freezing treatment is at a pressure in the range of about 0.1 MPa to about 55 MPa.

[0261] Aspect 67. The device of Aspect 64, wherein the isochoric freezing treatment is at a pressure in the range of about 0.1 MPa to about 25 MPa.

[0262] Aspect 68. The device of any one of Aspects 52-56, wherein the subfreezing temperature is -0.1 °C or less, the pressure is 1 MPa or more, and the treatment time is 30 minutes or more.

[0263] Aspect 69. The device of Aspect 68, wherein the subfreezing temperature is about -0.1 °C to -3.0 °C, the pressure is about 1 MPa to 35 MPa, and the treatment time is 48 hours or more.

[0264] Aspect 70. The device of Aspect 69, wherein the subfreezing temperature is about -2.5 °C to -3.0 °C, the pressure is about 25 MPa to 30 MPa and the treatment time is about 48 hours to 72 hours.

[0265] Aspect 71. The device of Aspect 70, wherein the subfreezing temperature is about -3.0 °C, the pressure is about 30 MPa and the treatment time is about 72 hours.

[0266] Aspect 72. The device of any one of Aspects 52-67, wherein the device enables opening the shells of the shellfish and / or detaching a meat from the shells.

[0267] Aspect 73. The device of any one of Aspects 68-71, wherein the device enables partially opening the shells of the shellfish and / or partially detaching a meat from the shells, thereby retaining liquor and avoiding meat dehydration.

[0268] Aspect 74. The device of Aspects 52-73, wherein the device is suitable for bulk processing.

[0269] Aspect 75. The device of Aspect 74, wherein the volume of the isochoric chamber is about 50 L to 10,000 L.

[0270] Aspect 76. The device of Aspect 74, wherein the volume of the isochoric chamber is about 1,000 L to 2,000 L.

[0271] Aspect 77. The device of any one of Aspects 52-76, wherein the isochoric chamber contains a nucleating agent.Attorney Docket: BCHR-002WO

[0272] Aspect 78. The device of any one of Aspects 52-77, wherein the fluid in the isochoric chamber is an aqueous solution or water.EXAMPLES

[0273] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperature, etc.) but some experimental errors and deviations should be accounted for.

[0274] All publications and patent applications cited in this specification are incorporated by reference herein as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference.

[0275] The present invention has been described in terms of particular embodiments found or proposed by the present inventor to comprise preferred modes for the practice of the invention. It will be appreciated by those of skill in the art that, in light of the present disclosure, numerous modifications and changes can be made in the particular embodiments exemplified without departing from the intended scope of the invention.Example 1. Opening up blue mussel shells and detach the meat from the shell by subjecting mussels to isochoric freezing at -2 °C

[0276] Blue mussels were packed into isochoric chambers and subjected to isochoric freezing at -2 °C for 96 hours, where 26 MPa pressure was created within the chamber. The blue mussels were alive when they went into the chamber and were closed shut. Some mussels were vacuum packed before going into the chamber, while others were put straight into the chamber without vacuum packing. After the isochoric freezing period, all the shells were open and the meat completely free from the shell, i.e. the meat simply dropped out of the shell all by itself (FIG. 7). This applied regardless of if the mussels were vacuum pack or not.Attorney Docket: BCHR-002WOExample 2. Opening up blue mussel shells and detaching meat from the shells by subjecting mussels to isochoric freezing at -6 °C

[0277] Blue mussels were packed into isochoric chambers and subjected to isochoric freezing at -6 °C for 48 hours, where 65 MPa pressure was created within the chamber. The blue mussels were alive when they went into the chamber and were closed shut, where opening them up manually required time and effort. Furthermore, the meat that was completely stuck to the whole inside of the shell and very difficult to scrape out, as demonstrated when a few mussels from the batch were forced open (FIG. 8A). Some mussels were vacuum packed before going into the chamber, while others were put straight into the chamber without vacuum packing. After the isochoric freezing period, all the shells were open and the meat completely free from the shell, i.e. the meat simply dropped out of the shell all by itself (FIG. 8B). This applied regardless of if the mussels were vacuum pack or not.Example 3. Opening up oysters and detaching the meat from the shell by subjecting the oysters to isochoric freezing

[0278] Oysters were packed into isochoric chambers and subjected to isochoric freezing at -7°C for 72 hours, where 75 MPa pressure was created within the chamber. The oysters were alive when they went into the chamber and were closed shut and proved impossible to open manually without tools (FIG. 9A). Furthermore, once an oyster from the batch had been forced open with tools, the meat that was firmly stuck to the inside of the shell in one place and somewhat difficult to remove from the shell (FIG. 9B). Some oysters were vacuum packed before going into the chamber, while others were put straight into the chamber without vacuum packing. After the isochoric freezing period, the shells of the vacuum packed oysters were open and the meat completely free from the shell, i.e. the meat simply dropped out of the shell all by itself (FIG. 9C). In the case of the oysters that were not vacuum packed, about half of them opened easily and the other half stayed shut.Example 4. Partial Shucking via Mild Isochoric Freezing

[0279] In some aspects of the present disclosure, certain pressures, significantly lower than those used in high-pressure processing, can lead to partial shell opening, enabling fasterAttorney Docket: BCHR-002WO shucking with a manual or mechanical method in the next step, and thereby leading to monetary gain.

[0280] This was demonstrated in experiments on different types of oysters. All conditions were kept under 35 MPa, which simplifies scaling for practical use down the line and minimizes the risk of any damage to the shellfish meat and its attributes.

[0281] Under subfreezing conditions with pressures at or below 35 MPa for storage periods exceeding 48 hours, oysters developed front-facing cracks in the shell, allowing easy insertion of a shucking knife and possible hand opening. These cracks occurred without causing full shell opening, and the adductor muscle was weakened to the extent that the meat could be removed cleanly with minimal force, preserving sensory properties and presentation.

[0282] One unexpected but commercially significant discovery was the retention of oyster liquor following mild isochoric freezing. In contrast, high-pressure processing (HPP) typically causes full shucking, resulting in the oysters gaping open and allowing the liquor to escape. To prevent this, producers often apply rubber bands to the oysters prior to HPP treatment, ensuring the shells remain closed post-process to retain the liquor and avoid meat dehydration.

[0283] The partial shucking method according to embodiments of the present invention circumvents this complication. Because the oysters only develop a controlled anterior crack without fully opening, they retain their natural liquor without requiring rubber bands or other interventions. This not only enhances flavor and freshness but also simplifies the processing workflow, reducing handling time and improving product presentation.

[0284] After treatment, the oyster meat was evaluated for sensory properties, such as texture and hardness, and was found to be indistinguishable from fresh hand-shucked raw oysters. This balance — enabling mechanical access while maintaining raw-like quality — addresses one of the major limitations of conventional HPP, which can alter texture due to excessive pressure.

[0285] These outcomes were consistent across both Pacific and Atlantic oysters.Quantitative ResultsAttorney Docket: BCHR-002WO

[0286] To better characterize the effectiveness of these conditions, the following table summarizes the shucking rates observed at different storage durations and pressure / temperature combinations:Table 1.

[0287] FIG. 10 shows the relationship between storage time and partial shucking efficiency. The shaded area represents the range of partial shucking observed in each condition. Labels denote pressure and temperature used. Results show a progressive increase in shucking percentage with time and pressure, peaking at 100% at 72 hours with 30 MPa and - 3°C.

[0288] For example, FIG. 11A shows Atlantic oysters after 48 hours of storage at 25 MPa and -2.5°C. The front of the shell shows a visible crack suitable for easy knife insertion. FIG. 11B shows Pacific oysters in the same condition (25 MPa, -2.5°C, 48 hours). Partial opening is evident while maintaining shell integrity. FIG. 11C shows that meat can be removed cleanly with minimal force due to weakening of the adductor muscle. Liquor remains inside, preserving flavor and presentation. FIG. 11D shows Pacific oysters at the same condition (30 MPa, -3°C, 72 hours). Partial opening is evident while maintaining shell integrity. FIG. HE shows an oyster with clean detachment of the adductor muscle, illustrating ease of removal and quality of presentation.

[0289] Collectively, these findings validate a novel use of mild isochoric freezing to achieve controlled shell weakening, enabling shucking without compromising meat quality or requiring additional processing aids. Also, the mild isochoric freezing of the present disclosure provides benefit of preserving flavor and presentation.

[0290] While the present invention has been described with reference to the specific embodiments thereof, it should be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit andAttorney Docket: BCHR-002WO scope of the invention. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, process, process step or steps, to the objective, spirit and scope of the present invention. All such modifications are intended to be within the scope of the claims appended hereto.

Claims

Attorney Docket: BCHR-002WOCLAIMSWhat is claimed is:

1. A method for processing shellfish comprising: a) placing shellfish in an isochoric chamber filled with a fluid; and b) isochoric freezing the fluid in the isochoric chamber at a subfreezing temperature over a treatment time; wherein a meat inside the shellfish is detached through the isochoric freezing.

2. The method of claim 1, further comprising c) warming the isochoric chamber containing the shellfish.

3. The method of claim 1 or 2, wherein the treatment time is 1 hour to 96 hours.

4. The method of any one of claims 1-3, wherein the treatment time is 1 hour to 48 hours.

5. The method of claim 4, wherein the treatment time is from 1 hour to 24 hours.

6. The method of any one of claims 1-5, wherein the subfreezing temperature is in the range of about 0 °C to -20 °C.

7. The method of claim 6, wherein the subfreezing temperature is in the range of about 0 °C to -10 °C.

8. The method of claim 6, wherein the subfreezing temperature is in the range of about 0 °C to -5 °C.

9. The method of claim 6, wherein the subfreezing temperature is in the range of about 0 °C to -2 °C.Attorney Docket: BCHR-002WO10. The method of any one of claims 1 to 9, wherein the isochoric freezing is at a pressure in the range of about 0.1 MPa to about 150 MPa.

11. The method of claim 10, wherein the isochoric freezing is at a pressure in the range of about 0.1 MPa to about 100 MPa.

12. The method of claim 10, wherein the isochoric freezing is at a pressure in the range of about 0.1 MPa to about 55 MPa.

13. The method of claim 10, wherein the isochoric freezing is at a pressure in the range of about 0.1 MPa to about 25 MPa.

14. The method of any one of claims 1-13, wherein the method comprises placing a large number of shellfish in the isochoric chamber for bulk processing.

15. The method of claim 14, wherein volume of the isochoric chamber is suitable for bulk processing.

16. The method of claim 15, wherein the volume of the isochoric chamber is about 50L to 10,000 L.

17. The method of claim 15, wherein the volume of the isochoric chamber is about 1,000 L to 2,000 L.

18. The method of any one of claims 1-17, wherein the shellfish is placed directly in the isochoric chamber.

19. The method of any one of claims 1 to 17, wherein the shellfish is placed in a matter container, and the matter container is placed in the fluid in the isochoric chamber.Attorney Docket: BCHR-002WO20. The method of any one of claims 1-17, wherein the shellfish is vacuum packed and placed in the isochoric chamber.

21. The method of any one of claims 1-20, wherein the isochoric chamber contains a nucleating agent.

22. The method of any one of claims 1-21, wherein the fluid in the isochoric chamber is an aqueous solution or water.

23. The method of any one of claims 1-22, wherein the shellfish is alive prior to the isochoric freezing.

24. The method of any one of claims 1-23, wherein the shellfish is raw shellfish or undercooked shellfish.

25. The method of any one of claims 1-24, wherein the shellfish is a bivalve, a univalve, a crustacean, a gastropod, a scaphopod, a polyplacophoran, or an echinoderm.

26. The method of claim 25, wherein the bivalve is selected from a mussel, a scallop, a clam, a winkle, and an oyster.

27. The method of claim 25, wherein the univalve is a snail.

28. The method of claim 25, wherein the crustacean is selected from a shrimp, a prawn, a crab, a lobster, barnacles, and a crayfish.

29. The method of claim 25, wherein the gastropod is selected from a conch, an abalone, a limpet, and a whelk.Attorney Docket: BCHR-002WO30. The method of any one of claims 1-29, wherein 30% or more, 50% or more, 70% or more, or 90% or more of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing, wherein the remaining shellfish are closed or partially opened.

31. The method of claim 30, wherein all of the shellfish are opened and the meat inside the shellfish are detached through the isochoric freezing.

32. The method of any one of claims 1-31, wherein the connective tissue of the shellfish to the shells is compromised by joint effects of subfreezing temperature and pressure under the conditions of isochoric freezing.

33. The method of any one of claims 1-32, wherein the isochoric freezing reduces or inhibits enzymatic activity in the shellfish and weakens adhesion of the shellfish to its shells.

34. A method for partially opening shellfish comprising: a) placing shellfish in an isochoric chamber filled with a fluid; and b-2) isochoric freezing the fluid in the isochoric chamber with a subfreezing temperature of -0.1 °C or less, a pressure of 1 MPa or more, and a treatment time of 30 minutes or more.

35. The method of claim 34, comprising c) warming the isochoric chamber containing the shellfish.

36. The method of claim 34 or 35, wherein the subfreezing temperature is about -0.1 °C to - 3.0 °C, the pressure is 1 MPa to 35 MPa, and the treatment time is 48 hours or more.

37. The method of claim 36, wherein the subfreezing temperature is about -2.5 °C to -3.0 °C, the pressure is about 25 MPa to 30 MPa and the treatment time is about 48 hours to 72 hours.Attorney Docket: BCHR-002WO38. The method of claim 37, wherein the subfreezing temperature is about -3.0 °C, the pressure is about 30 MPa and the treatment time is about 72 hours.

39. The method of any one of claims 34-38, wherein the method comprises placing a large number of shellfish in the isochoric chamber for bulk processing.

40. The method of any one of claims 34-39, wherein the shellfish is placed directly in the isochoric chamber.

41. The method of any one of claims 34-39, wherein the shellfish is placed in a matter container, and the matter container is placed in the fluid in the isochoric chamber.

42. The method of any one of claims 34-39, wherein the shellfish is vacuum packed and placed in the isochoric chamber.

43. The method of any one of claims 34-42, wherein the isochoric chamber contains a nucleating agent.

44. The method of any one of claims 34-43, wherein the fluid in the isochoric chamber is an aqueous solution or water.

45. The method of any one of claims 34-44, wherein the shellfish is alive prior to the isochoric freezing.

46. The method of any one of claims 34-44, wherein the shellfish is raw shellfish or undercooked shellfish.

47. The method of any one of claims 34-46, wherein the shellfish is a bivalve, a univalve, a crustacean, a gastropod, a scaphopod, a polyplacophoran, or an echinoderm.Attorney Docket: BCHR-002WO48. The method of claim 47, wherein the bivalve is selected from a mussel, a scallop, a clam, a winkle, and an oyster.

49. The method of claim 47, wherein the crustacean is selected from a shrimp, a prawn, a crab, a lobster, barnacles, and a crayfish.

50. The method of claim 47, wherein the gastropod is selected from a conch, an abalone, a limpet, and a whelk.

51. The method of claims 34-50, wherein the shellfish remains mostly closed but is structurally weakened, thereby retaining liquor and avoiding meat dehydration.

52. A device for processing shellfish comprising: a) an isochoric chamber filled with a fluid, wherein the isochoric chamber contains shellfish; and b) a temperature control system configured to cool the fluid in the isochoric chamber containing the shellfish to a subfreezing temperature over a treatment time and warm the fluid in the isochoric chamber containing the shellfish.

53. The device of claim 52, wherein the temperature control system comprises a cooling system and a heating system configured to cool and warm the fluid in the isochoric chamber containing the shellfish.

54. The device of claim 52 or 53, wherein the device comprises a pressure monitoring element.

55. The device of any one of claims 52-54, wherein the device comprises a pressure relief mechanism.Attorney Docket: BCHR-002WO56. The device of any one of claims 52-55, wherein the device comprises multiple discrete sub-systems for physically separating shellfish from ice.

57. The device of any one of claims 52-56, wherein the treatment time is 1 hour to 96 hours.

58. The device of claim 57, wherein the treatment time is 1 hour to 48 hours.

59. The device of claim 57, wherein the treatment time is from 1 hour to 24 hours.

60. The device of any one of claims 52-59, wherein the subfreezing temperature is in the range of about 0 °C to -20 °C.

61. The device of claim 60, wherein the subfreezing temperature is in the range of about 0 °C to -10 °C.

62. The device of claim 60, wherein the subfreezing temperature is in the range of about 0 °C to -5 °C.

63. The device of claim 60, wherein the subfreezing temperature is in the range of about 0 °C to -2 °C.

64. The device of any one of claims 52-63, wherein the isochoric freezing treatment is at a pressure in the range of about 0.1 MPa to about 150 MPa.

65. The device of claim 64, wherein the isochoric freezing treatment is at a pressure in the range of about 0.1 MPa to about 100 MPa.

66. The device of claim 64, wherein the isochoric freezing treatment is at a pressure in the range of about 0.1 MPa to about 55 MPa.Attorney Docket: BCHR-002WO67. The device of claim 64, wherein the isochoric freezing treatment is at a pressure in the range of about 0.1 MPa to about 25 MPa.

68. The device of any one of claims 52-56, wherein the subfreezing temperature is -0.1 °C or less, the pressure is 1 MPa or more, and the treatment time is 30 minutes or more.

69. The device of claim 68, wherein the subfreezing temperature is about -0.1 °C to -3.0 °C, the pressure is about 1 MPa to 35 MPa, and the treatment time is 48 hours or more.

70. The device of claim 69, wherein the subfreezing temperature is about -2.5 °C to -3.0 °C, the pressure is about 25 MPa to 30 MPa and the treatment time is about 48 hours to 72 hours.

71. The device of claim 70, wherein the subfreezing temperature is about -3.0 °C, the pressure is about 30 MPa and the treatment time is about 72 hours.

72. The device of claims 52-67, wherein the device enables opening the shells of the shellfish and / or detaching a meat from the shells.

73. The device of any one of claims 68-71, wherein the device enables partially opening the shells of the shellfish and / or partially detaching a meat from the shells, thereby retaining liquor and avoiding meat dehydration.

74. The device of any one of claims 52-73, wherein the device is suitable for bulk processing.

75. The device of claim 74, wherein volume of the isochoric chamber is about 50 L to 10,000 L.

76. The device of claim 74, wherein the volume of the isochoric chamber is about 1,000 L to 2,000 L.Attorney Docket: BCHR-002WO77. The device of any one of claims 52-76, wherein the isochoric chamber contains a nucleating agent.

78. The device of any one of claims 52-77, wherein the fluid in the isochoric chamber is an aqueous solution or water.

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