METHOD FOR PREPARING A COMPOSITION DERIVED FROM LIVE SEA CHURIES

Abstract

METHOD FOR PREPARING A COMPOSITION FROM LIVE SEA CUBES The invention relates to a method for preparing a composition from sea cucumbers, said method being characterized in that: (E1) at least two live sea cucumbers are selected; in a step (E2) of contact with fresh water, said sea cucumbers are placed in a container containing water having a maximum of 15 grams of dissolved salts per liter for a predetermined period of between 20 minutes and 4 hours, each sea cucumber being alive at the end of this step of contact with fresh water; at the end of said step of contact with fresh water, said sea cucumbers are removed (E3) from said container and said water contained in said container is recovered (E4), as a composition from live sea cucumbers; said composition thus recovered is filtered (E5); the filtrate is recovered (E6). Figure for the summary: Figure 1

Description

Title of the invention: PROCESS FOR PREPARING A COMPOSITION FROM LIVE SEA CHERRY Technical field of the invention

[0001] The invention relates to a process for preparing a composition derived from sea cucumbers. The invention is particularly intended for use in the cosmetic, pharmaceutical or nutraceutical field. In particular, the invention relates to such a process for preparing a composition derived from sea cucumbers that can be used as a cosmetic and / or pharmaceutical and / or nutraceutical active ingredient. Technological background

[0002] Sea cucumbers, or holothurians, are marine invertebrates found in many marine areas, generally near the bottom of seas and oceans.

[0003] Sea cucumbers play a key role in protecting coral reefs, particularly against the devastating effects of climate change. Sea cucumbers partially protect coral growth from the harmful effects of ocean acidification. Sea cucumber excrement, resulting from the natural digestion of the sand they ingest, raises the pH of reef waters, thus counteracting the impact of ocean acidification. When sea cucumbers digest sand, they release calcium carbonate (CaCO3), a key component in the constitution of corals. To survive, coral reefs must accumulate enough calcium carbonate to compensate for or neutralize the losses caused by erosion.Research shows that in a healthy reef system, the dissolution of calcium carbonate-rich sediments by sea cucumbers and other bioeroding organisms appears to be a key contributor to the natural renewal of carbonate substrates. In addition, the ammonia released by sea cucumbers enriches the surrounding waters and stimulates coral growth through this nutrient supply.

[0004] However, sea cucumbers are widely harvested and exploited throughout the world, particularly in Asia. They are consumed as a culinary delicacy and considered a traditional tonic remedy.

[0005] Document WO2005120185 describes for example the recovery of triterpene glycosides from freeze-dried tissues, spray-dried from the cooking water or reduced to dry powder, of sea cucumbers Cucumaria frondosa which have undergone industrial treatment by extraction using ethyl acetate and water, followed by chromatography in order to separate the triterpene glycosides. Such a process is complex and involves cooking and / or reducing the sea cucumbers as such to powder.

[0006] The inventors sought to propose a process for preparing a composition derived from sea cucumbers of interest for applications in the cosmetic, pharmaceutical or nutraceutical fields while avoiding threatening sea cucumbers and their natural ecosystem. Objectives of the invention

[0007] The invention aims to provide a process for preparing a composition derived from sea cucumbers which overcomes at least some of the drawbacks of known solutions.

[0008] The invention also aims to provide, in at least one embodiment, a method for preparing a composition derived from sea cucumbers which is not only sustainable and environmentally friendly but also avoids any suffering to the sea cucumbers.

[0009] The invention also aims to provide, in at least one embodiment, a method for preparing a composition derived from live sea cucumbers, which can be implemented for applications in the cosmetic field, in the pharmaceutical field and in the nutraceutical field.

[0010] The invention also aims to provide, in at least one embodiment, a composition capable of being used as a cosmetic and / or pharmaceutical and / or nutraceutical active ingredient. Statement of the invention

[0011] To this end, the invention relates to a method for preparing a composition derived from live sea cucumbers, said method being characterized in that:

[0012] - at least two live sea cucumbers are chosen;

[0013] - in a step, called the fresh water contact step, said ho are placed sea ​​cucumbers in a container containing water containing a maximum of 15 grams of dissolved salts per liter, for a predetermined period of between 20 minutes and 4 hours, each sea cucumber being alive at the end of this step of contact with fresh water;

[0014] - at the end of said step of contacting with fresh water, said sea ​​cucumbers from said container and said water contained in said container is recovered as a composition from live sea cucumbers.

[0015] Sea cucumbers lack any defenses visible to the naked eye. To defend themselves against predators in their natural environment, they synthesize defense molecules that they release into the water.

[0016] Surprisingly, the inventors have found that it is possible to determine the conditions for recovering compounds of interest released spontaneously by sea cucumbers without (irreversibly) altering their state of health. The water recovered at the end of said step of contacting with fresh water after removing the sea cucumbers therefore contains the molecules released by the living sea cucumbers. Indeed, in particular, the inventors have found that it is necessary not to exceed a duration of 4 consecutive hours in contact with water containing less than 15 grams of dissolved salts per liter. Thus, each sea cucumber is still alive at the end of the step of contact with fresh water, and in particular does not show any sign of deterioration in its state of health such as evisceration or whitening of the integument or the presence of ulcers on the integument of the sea cucumbers. A method according to the invention therefore makes it possible to prepare a composition derived from live sea cucumbers without requiring their sacrifice and therefore "humanely".

[0017] The health status of the sea cucumbers subjected to a method according to the invention, and in particular to a contacting step according to the invention, is therefore not permanently affected. This even makes it possible to subsequently reuse the same sea cucumber individuals again to repeat the method and therefore to repeat the contacting step with fresh water. The steps of a method according to the invention can therefore be repeated one or more times by reusing the same sea cucumbers (identical individuals). It is of course also possible to repeat all the steps of the method with different sea cucumbers (in whole or in part).

[0018] The reflex of release of the compounds of interest by the sea cucumbers is caused by contact with so-called fresh water, and by bathing, at least by their ventral part (in particular from the mouth to the anus), in such fresh water which is distinguished from the sea water ("salt") corresponding to their usual natural environment. Using water comprising less than 15 grams of dissolved salts per liter is sufficient to allow such a release of the desired compounds, the sea water in which the sea cucumbers can live can for example comprise of the order of 28 to 36 g / L of dissolved salts. When we speak of less than 15 grams of dissolved salts per liter, this refers to all salts combined (including sodium chloride NaCl). Water with a lower salt content can also be used, for example less than 8 grams of dissolved salts per liter or less than 4 grams of dissolved salts per liter.Advantageously and according to the invention, during said step of contacting with fresh water, water containing a maximum of 1 gram of dissolved salts per liter is used. Such a title may correspond to pure water or so-called osmosis water. Advantageously and according to the invention, during said step of contacting with fresh water, osmosis water is used. The use of such water having very few dissolved salts makes it possible to control the quality and nature of the composition obtained from live sea cucumbers, ensuring better reproducibility of the process according to the invention. Thus, advantageously and according to the invention, during said step of contacting with fresh water, water, called osmosis water, obtained by a reverse osmosis step and containing a maximum of 1 gram of dissolved salts per liter is used.

[0019] Several sea cucumbers can be placed together in the same container used for said step of contacting with fresh water. The volume of water used must be sufficient for each sea cucumber to be in contact with it, at least at the start of the contacting step, i.e. each sea cucumber bathes at least partly in said water without necessarily being entirely immersed in it. Advantageously and according to the invention, during said step of contacting with fresh water, each sea cucumber is entirely immersed in fresh water. Advantageously and according to the invention, during said step of contacting with fresh water, a volume of water at least equal to 1 liter of water is placed in each container for 30 sea cucumbers to be placed in said container. This makes it possible to avoid any discomfort or inconvenience between said sea cucumbers present in the same container.More particularly, during said step of contacting with fresh water, a volume of water at least equal to 1 liter of water for 5 to 25 sea cucumbers to be placed in said container is placed in each container. There is also nothing to prevent the provision of a container having several compartments or partitions capable of separating the sea cucumbers from each other, individually or not.

[0020] Advantageously and according to the invention, during said step of bringing into contact with fresh water, a number of sea cucumbers is placed in each container such that the mass of sea cucumber placed in 1 liter of water is between 50 grams and 10,000 grams (10 kg), in particular between 300 grams and 3,000 grams (3 kg), and in particular between 1,000 grams (1 kg) and 2,000 grams (2 kg).

[0021] The containers used may have any shape suitable for their use and allowing the introduction and removal (withdrawal) of the sea cucumbers at the beginning and end of said contacting step. Each container has at least one bottom and peripheral side walls. Each container may be at least partially open at the top (“open to the air”) or even completely closed (for example using a lid or in the manner of a box, hermetically or not).

[0022] The temperature of the water used may vary and may or may not be constant throughout said contacting step. In general, the water used is at room temperature. Advantageously and according to the invention, during said contacting step with fresh water, water at a temperature between 0°C and 50°C, in particular between 15°C and 35°C, is used.

[0023] All species of sea cucumbers can be used in a method according to the invention. It is possible to use sea cucumbers of the same species or of different species during said step of contacting with fresh water. Advantageously and according to the invention, each sea cucumber is chosen from the group formed by the species Holothuria fuscogilva (white teat sea cucumbers) and the species Ho-lothuria whitmaei (black teat sea cucumbers).

[0024] Advantageously and according to the invention, each sea cucumber is chosen such that each sea cucumber has a mass of between 5 grams and 5,000 grams (5 kg), in particular between 15 grams and 2,000 grams (2 kg), in particular such that each sea cucumber has a mass of between 100 grams and 250 grams.

[0025] Each sea cucumber used in a method according to the invention may be taken from a sample in the natural environment or preserved within an aquaculture farm and / or come from an aquaculture hatchery.

[0026] Advantageously and according to the invention, at the end of said step of bringing into contact with fresh water, said sea cucumbers removed from said container are placed in water containing at least 20 grams of dissolved salts per liter.

[0027] In order to best preserve the health of the sea cucumbers, it is useful to return them to an environment with conditions close to their natural living conditions (or identical) at the end of each step of contact with fresh water. Advantageously and according to the invention, after said step of contact in fresh water, each sea cucumber is left in water having at least 20 grams of dissolved salts per liter for at least 12 hours so as to allow each sea cucumber to be subjected to a possible new step of contact with fresh water. For example, a rest period of 24 hours or even 48 hours may be provided for said sea cucumbers between several successive steps of contact with fresh water in the context of a method according to the invention.

[0028] Advantageously and according to the invention, after removing the sea cucumbers and recovering the composition from live sea cucumbers, said composition thus recovered is filtered and the filtrate resulting from this filtration is recovered. The step of filtering the composition from live sea cucumbers recovered at the end of the step of bringing into contact with fresh water makes it possible to separate any possible debris or unwanted object such as grains of sand or fragments of algae possibly transported by the sea cucumbers. Any filtration technique can be used (sieve, filter paper, etc.) or even a separation technique by decantation and / or sedimentation.Advantageously and according to the invention, at least one step of filtration of the composition recovered at the end of said step of contacting with fresh water is carried out using a filtration device whose pores have an average diameter of between 0.45 pm and 800 pm, in particular between 40 pm and 600 pm.

[0029] The composition obtained from live sea cucumbers after filtration can be preserved as is or concentrated or dried by any suitable technique such as freeze-drying. Thus, advantageously and according to the invention, after having recovered said filtrate, said filtrate is freeze-dried.

[0030] The composition from live sea cucumbers obtained after filtration can be stored, for example away from light, for a period of several days at several months, or even several years, with a view to its use as a cosmetic and / or pharmaceutical and / or nutraceutical active ingredient.

[0031] A method according to the invention respecting the sea cucumbers and not causing any irreversible effect on their state of health, it is entirely possible and particularly advantageous to be able to reuse the same sea cucumbers several times. According to an alternative embodiment, after having recovered said composition from live sea cucumbers, said steps are repeated starting from said step of bringing into contact with fresh water.

[0032] The invention also relates to a method and a composition, characterized in combination by all or part of the characteristics mentioned above or below. List of figures

[0033] Other aims, characteristics and advantages of the invention will appear on reading the following description given solely for non-limiting purposes and which refers to the appended figure in which:

[0034] [Fig-1] is a schematic view of a method according to a first embodiment of the invention.

[0035] Detailed description of an embodiment of the invention

[0036] A method of preparing a composition derived from sea cucumbers comprises the following steps illustrated in [Fig.l]:

[0037] - in a first step E1 at least two live sea cucumbers are chosen;

[0038] - in a second step E2 of contacting in fresh water, the ho are placed sea ​​cucumbers in a container containing water containing a maximum of 15 grams of dissolved salts per litre for a predetermined period of between 20 minutes and 4 hours, each sea cucumber being alive and with no sign of significant deterioration in its state of health at the end of this stage of contact with fresh water;

[0039] - at the end of the step of contacting with fresh water, in a third step E3, each sea cucumber is removed from the container and in a fourth step E4, the water contained in said container is recovered as a composition comprising a composition derived from live sea cucumbers;

[0040] - in a fifth step E5, the composition recovered in step E4 is filtered;

[0041] - then, in a sixth step E6, the filtrate from the filtration carried out at step E5,

[0042] - in an optional seventh step E7, a new filtration of the filtrate is carried out recovered in step E6, and

[0043] - in an optional eighth step E8, the filtrate from the filtration is recovered operated in step E7.

[0044] Using water containing less than 15 grams of dissolved salts per liter is sufficient to allow such a release of the desired compounds. It is also possible to use water having a lower salt content, for example less than 8 grams of dissolved salts per liter or less than 4 grams of dissolved salts per liter. For example, water containing a maximum of 1 gram of dissolved salts per liter is used. Such a content can correspond to pure water or so-called osmosis water.

[0045] A number of sea cucumbers are placed in each container such that the mass of sea cucumber placed in 1 liter of water is between 50 g and 10 kg, in particular between 300 g and 3 kg, and in particular between 1 kg and 2 kg. Each sea cucumber has a mass of between 5 g and 5 kg, in particular between 15 g and 2 kg, between 25 g and 1 kg and more particularly a mass of between 100 g and 250 g.

[0046] It is possible to repeat steps E1 to E4, or E1 to E6 or E1 to E8, after having left the sea cucumbers to stand in sea water (or water containing at least 20 grams of dissolved salts per liter) for at least a few hours (for example 12 to 48 hours).

[0047] EXAMPLE 1

[0048] 40 juvenile sea cucumbers aged 20 months and belonging to the species Ho-lothuria fuscogilva were chosen. These juvenile sea cucumbers were produced in a hatchery and then transferred to a nursery (in tanks on land) located at the Ifremer Pacific center in Vairao (Tahiti, French Polynesia). Following the nursery stage, these sea cucumbers are transferred to sea pens for the pre-growing and grow-out phases. These sea pens allow these sea cucumbers to evolve at a depth of between 0.50 m and 1.50 m below sea level.

[0049] There are 4 containers each with a capacity of 1.5 L. Each harvested sea cucumber is placed directly in one of the 4 containers without cleaning them (i.e. without removing any sand that may be present on their bodies so as not to stress them). 10 sea cucumbers are placed per container. One liter of osmosis water (at 25°C) is poured into each container. Each container is a plastic tub closed with a lid after adding the osmosis water to each tub at the start of the contact stage with fresh water.

[0050] The table below indicates the duration for which the sea cucumbers were placed in contact with osmosis water and their weight measured at the end of the step of placing them in contact with osmosis water.

[0051] The sea cucumbers are removed from each tank at the end of the freshwater contact step and the remaining composition is recovered from each tank. Each recovered composition is filtered using a filter screen having a 500 μm mesh size and each collected filtrate is stored. Each live sea cucumber composition thus collected has a brown color. Each live sea cucumber composition is stored in a leak-proof flexible bag in a freezer at -18°C.

[0052] The general condition of the sea cucumbers is monitored for 3 days following the end of immersion in reverse osmosis water. To do this, they are kept in stabling tanks for 3 days before being returned to their original enclosure.

[0053] [Tableauxl] Duration of the bath in osmosis water (in hours) Total weight of the 10 sea cucumbers (in grams) Average weight (in grams) Survival after 3 days Tank No. 1 1,804 80.40 100% Tank No. 2 2,981 98.10 100% Tank No. 3 3,1085 108.50 100% Tank No. 4 4,786 78.60 100%

[0054] It is therefore observed that such a process makes it possible to obtain a composition comprising compounds of interest released by the sea cucumbers, not only without causing their death (neither during the step of bringing them into contact with fresh water itself, nor as a result thereof), but also without harming their state of health. The sea cucumbers used can be reused subsequently to repeat the same steps of the process.

[0055] EXAMPLE 2

[0056] 40 juvenile sea cucumbers aged 21 months and belonging to the species Ho-lothuria fuscogilva were chosen. These juvenile sea cucumbers were produced in a hatchery and then transferred to a nursery (in tanks on land) located at the Ifremer Pacific center in Vairao (Tahiti, French Polynesia). Following the nursery stage, these sea cucumbers are transferred to sea pens for the pre-growing and grow-out phases. These sea pens allow these sea cucumbers to evolve at a depth of between 0.50 m and 1.50 m below sea level.

[0057] A stainless steel container with a capacity of 10.18 L is provided. Each harvested sea cucumber is placed directly in the container after having gently cleaned them to remove any algae and sand that may be attached to them. The 40 sea cucumbers are placed in the container (stainless steel tank) by placing them next to each other and avoiding stacking them on top of each other. 4 liters of previously osmosis water (at 25°C) are poured into the container before placing a lid closing the opening of the container. At the start of the contact stage with fresh water, each sea cucumber is completely submerged. The sea cucumbers are left in the osmosis water for 3 hours (protected from ultraviolet radiation, taking care to avoid any other external disturbance).

[0058] The sea cucumbers are removed from the stainless steel tank at the end of the plating step. contact in fresh water and the remaining composition is recovered. The recovered composition is filtered using a first filter sieve with a mesh size of 100 μm, followed by a second filter sieve with a mesh size of 41 μm and the collected filtrate is stored in a glass jar. Each composition from live sea cucumbers is directly frozen in stainless steel trays in a freezer at -10°C and freeze-dried.

[0059] EXAMPLE 3

[0060] 30 juvenile sea cucumbers with an average mass of 84 grams and belonging to the species Holothuria fuscogilva were chosen. These sea cucumbers came from the same hatchery and lived in the same conditions as those of the previous examples 1 and 2.

[0061] There are 3 containers each with a capacity of 1.5 L. There are 10 sea cucumbers per container. In each container, a different quantity of osmosis water is added (IL (tank no. 5), 0.75 L (tank no. 6) and 0.5 L (tank no. 7)). Each container is a plastic container closed with a lid after adding the osmosis water to each tank at the start of the contacting step. The duration of the contacting step in fresh water is 3 hours.

[0062] The sea cucumbers are removed from each tank at the end of the freshwater contact step and the remaining composition is recovered from each tank. Each recovered composition is filtered using a filter screen with a mesh size of 41 μm and each collected filtrate is stored. Each composition from live sea cucumbers is directly frozen in a stainless steel tray in a freezer at -20°C for 24 hours and then freeze-dried using a freeze-dryer marketed by the company CRYOTECH (Saint-Gély-du-Fesc, France) under the reference COSMOS® and having a capacity of 3 kg.

[0063] The volumes of compositions from filtered live sea cucumbers are respectively 1.10 L, 0.78 L and 0.55 L for tanks No. 5, 6 and 7. The masses of freeze-dried compositions are respectively 9.36 g, 11.05 g and 7.78 g for tanks No. 5, 6 and 7. We therefore observe an increase in yield proportional to the increase in the number of sea cucumbers per liter of fresh water (osmosed here).

[0064] The general condition of the sea cucumbers is monitored for 24 hours following the end of immersion in the osmosis water. To do this, they are kept in a tank before being returned to their original enclosure.

[0065] Turning tests are carried out immediately after the contact step in the osmosis water. They make it possible to assess the state of health of each sea cucumber, a sea cucumber placed on its dorsal part having the reflex to turn over in order to be ventral part towards the bottom of the container in which it is located.

[0066] The following table groups together the durations measured until the complete reversal of each sea cucumber.

[0067] For comparison, the turning time was also measured for 10 sea cucumbers that had not undergone a prior step of contact with fresh water. This average time is 12 minutes (with a standard deviation between the 10 measurements equal to 14). For comparison, the turning time was also measured for 10 sea cucumbers in one liter of osmosis water after 1 minute of contact and 1 hour of contact. This average turning time is 19 minutes (with a standard deviation between the 10 measurements equal to 17) after 1 minute of contact, and the average turning time is 60 minutes (with a standard deviation between the 10 measurements equal to 30) after 1 hour of contact.

[0068] These measurements show that the freshwater contact step lengthens the turning time of the sea cucumbers but does not inhibit it. The turning of the sea cucumbers is therefore only delayed by contact with freshwater for a period of less than 4 hours. These results also show that an increasing number of sea cucumbers per liter of water (10 sea cucumbers per 1 liter to 20 sea cucumbers per 1 liter of osmosis water) does not negatively impact this turning reflex.

[0069] [Tables2] Container Tank No. 5 Tank No. 6 Tank No. 7 Time until sea cucumber re-turning in minutes 35.83 29.50 28.58 36.43 31.00 31.00 47.17 33.33 31.00 49.67 40.15 33.50 63.68 41.58 35.83 64.43 43.17 42.43 64.83 45.08 43.98 67.10 48.73 47.17 74.23 49.00 51.98 56 56.78 55.82 Average time in minutes 56 40 38 Standard deviation 14 9 10

[0070] Histological analyses of the integument of podias of the dorsal and ventral parts of sea cucumbers (after 1 hour of contact in a liter of osmosis water and those of tanks No. 5, No. 6 and No. 7) were also carried out. The integument of the podias is composed of five superimposed layers, of which the cuticle, which is acellular, constitutes the outermost layer. Underneath the cuticle is a cellular layer forming an epithelium. At the dorsal level, the subepithelium is distinguished by large empty alveoli (ossicula lacunae) following decalcification treatment. We then find two merged laminar layers which are differentiated by their density, of which the densest layer is located under the subepithelium. We find, whether in the dorsal or ventral integument, brown pigments distributed in a non-homogeneous manner in the epithelium and in the subepithelium. Unlike the dorsal integument, the ventral part contains fewer ossicula lacunae and more secretory glands in the subepithelium. The glands are characterized by an alignment of nuclei delimiting this secretory structure with a higher concentration of nuclei on the cuticle.

[0071] From 1 hour of contact in a liter of osmosis water, a detachment of the cuticle is observed on the dorsal and ventral integuments. Associated with this, a lysis of the epithelium cells is also observed. In the ventral integument, an atrophy of the glands located in the sub-epithelium can be seen in the treatments corresponding to tank No. 6 and tank No. 7. However, the morphology of the laminar zones (dense and diffuse) is not affected by contact with the osmosis fresh water.

[0072] Thus, contact with fresh water causes detachment of the cuticle of the integument and lysis of the epithelium cells, from 1 hour of treatment without affecting the deep layers of the integument and this is also true for contact for 3 hours at different densities of sea cucumbers per liter of osmosis water. It is concluded that the treatments only stress the integument superficially and show that the defense molecules contained in the integument are released by the lysis of the epithelium cells.

[0073] Taking into account the turning reflex, the recovery rate 24 hours after contact and the condition of the integument, it was found that the stress suffered by the juveniles treated by a method according to the invention is superficial. A 1-hour treatment effectively slows down the turning reflex but does not abolish it. A similar evisceration rate is found between juveniles bathed in osmosis water or seawater.

Claims

Claims

1. Process for preparing a composition derived from live sea cucumbers, said process being characterized in that it comprises the following steps: • (E1) at least two live sea cucumbers are chosen; • in a step, called step (E2) of contacting with fresh water, said sea cucumbers are placed in a container containing water having a maximum of 15 grams of dissolved salts per liter, for a predetermined period of between 20 minutes and 4 hours, each sea cucumber being alive at the end of this step of contacting with fresh water; • at the end of said step of contacting with fresh water, (E3) said sea cucumbers are removed from said container and (E4) said water contained in said container is recovered as a composition derived from live sea cucumbers.

2. Method according to claim 1, characterized in that it further comprises the following steps: • (E5) said composition recovered in step (E4) is filtered; • (E6) said filtrate is recovered.

3. Method according to one of claims 1 or 2, characterized in that during said step of contacting with fresh water, water is used, called osmosis water, obtained by a reverse osmosis step and containing a maximum of 1 gram of dissolved salts per liter.

4. Method according to one of claims 1 to 3, characterized in that during said step (E2) of contacting with fresh water, a volume of water at least equal to 1 liter of water for 30 sea cucumbers to be placed in said container is placed in each container.

5. Method according to one of claims 1 to 4, characterized in that during said step (E2) of contacting with fresh water, water at a temperature between 0°C and 50°C is used, in particular water at a temperature between 15°C and 35°C.

6. Method according to one of claims 1 to 5, characterized in that each sea cucumber is chosen from the group formed by the species Holothuria

7.

8.

9.

10.

11. fuscogilva and the species Holothuria whitmaei. Method according to one of claims 1 to 6, characterized in that each sea cucumber is chosen such that each sea cucumber has a mass of between 5 grams and 5,000 grams, in particular such that each sea cucumber has a mass of between 100 grams and 250 grams. Method according to one of claims 1 to 7, characterized in that at the end of said step (E2) of bringing into contact with fresh water, said sea cucumbers removed from said container are placed in water containing at least 20 grams of dissolved salts per liter. Method according to claim 8, characterized in that after said step (E2) of contacting with fresh water, each sea cucumber is left in water containing at least 20 grams of dissolved salts per liter for at least 12 hours. Method according to claim 2, characterized in that after having recovered said filtrate, said filtrate is lyophilized. Method according to one of claims 1 to 10, characterized in that after having recovered said composition from live sea cucumbers, said steps are repeated starting from said step (E2) of contacting with fresh water.