Precious coral cultivation method by fusion

Abstract

The present invention relates generally to the field of precious coral cultivation and propagation, and more particularly to a method for producing or culturing precious corals, or colonies, recruits, or chimeras of precious corals from fragments, existing colonies, or recruits of precious corals belonging to the Corallinaceae family.

Description

[Technical Field]

[0001] The present invention relates generally to the field of precious coral cultivation and propagation, and more particularly to a method for producing or culturing precious corals, or colonies, recruits, or chimeras of precious corals from fragments, colonies, or existing recruits of precious corals belonging to the Corallinaceae family. [Background technology]

[0002] Since the Neolithic period, precious coral has been used as amulets, jewelry, trading currency, and in many cultures as a magical symbol, religious symbol, symbol of good luck and social status due to its aesthetic qualities. Since Roman times, precious coral has been valued for its medicinal properties and is still used today in homeopathic medicine and traditional Chinese medicine.

[0003] According to the World Jewellery Confederation (CIBJO), of the more than 6,000 species of Cnidaria, class Anthozoa, commonly known as corals, only eight species are defined as gem corals used in fine jewellery. These species all belong to the family Corallidae, suborder Scleractinia, order Alcyonacea, and subclass Octocorallia.

[0004] Precious corals not only have the density and hardness of their carbonate skeletons, which makes them extremely attractive for jewelry, but also have characteristics that make them significantly different from typical coral "reefs," particularly in their ecology. Unlike typical coral reefs, precious corals live in the mid-depth zone and aphotic zone of the deep sea (50m to 2000m). In fact, because these corals are not in a symbiotic relationship with the microalgae zooxanthellae, they do not need exposure to sunlight to carry out photosynthesis.

[0005] Precious corals are long-lived corals, but they grow slowly and have low reproductive ability. In addition, their larvae have low dispersal ability, making them highly vulnerable to overfishing.

[0006] In fact, years of overfishing have dramatically changed the population structure of precious corals. In shallower waters, in particular, fishing rates are generally high, and large colonies of precious corals have almost disappeared. Coral "forests" have been transformed into "grasslands" of young colonies with poorly developed three-dimensional structures, which no longer constitute functional habitats.

[0007] Temperate coral ecosystems are rich in biodiversity, and their ongoing disappearance poses an important problem from an ecological perspective. In order to address this critical issue, several restrictions have been imposed on the collection of precious corals, and four of the eight most valuable species are listed as "endangered species" in Appendix III (species whose trade is regulated at the regional level) created by the Convention on International Trade in Endangered Species of Flora and Fauna (CITES).

[0008] Given that the coral industry is an important area for many local communities and must face the risk of economic extinction of precious corals (FAO data shows that catches of precious corals in the Mediterranean Sea fell by 95% between 1974 and 2020), the ongoing disappearance of precious coral colonies is also an important issue from a socio-economic point of view.

[0009] To ensure greater protection, China and the United States have submitted two formal requests to add eight valuable coral species to CITES Appendix II (species whose international trade is restricted). However, such a measure was strongly opposed by the sector's major producer countries, as it would risk a rapid decline in the precious coral industry and have serious impacts on the local communities that commercially exploit these species.

[0010] To address the ongoing decline in wild populations, many marine protected areas (MPAs) have been established in recent years for the passive conservation of precious corals. However, because these species require long periods of time to grow and the recovery of their three-dimensional structure, even within the same MPAs, it has become clear that passive conservation alone is insufficient to address increasingly frequent mass mortality events (MMEs).

[0011] Therefore, there are many challenges in managing the available resources of precious corals. On the one hand, biologists are calling for the adoption of stricter regulations regarding coral harvesting, proposing the establishment of Marine Protected Areas (MPAs) and the listing of all coral species in Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES), which could result in the risk of a contraction of the industry operating in this field. On the other hand, the coral industry is seeking permission to use remotely operated underwater vehicles to explore and extract currently unavailable coral reefs, which could result in the risk of the biological extinction of these coral species.

[0012] Corals of the Staghorn coral family have long growth periods and specialized biology. Currently, it is not feasible to develop effective, large-scale strategies for cultivating these species. In fact, all culture trials employed to date have proven ineffective or economically unprofitable. Therefore, wild coral collection remains the only source of supply in this field.

[0013] As mentioned above, precious corals grow very slowly, taking several decades to reach a basal diameter of 7 mm (i.e., the minimum size allowed for harvest in the Mediterranean Sea as determined by the General Fisheries Commission for the Mediterranean Sea (GFCM)). Even after that, they require a very long cultivation period, making them economically unsustainable.

[0014] All currently employed active repopulation strategies for precious corals involve direct transplantation of coral colonies collected from the wild (Villechanoux et al., Water 2022, 14(7), 1071; Koido et al., Zoological Studies 2022, 61:46). This type of repopulation strategy, which does not involve the prior step of culturing corals in a propagating nursery, has already been demonstrated (for common corals) to be less effective in increasing total coral biomass. Initial results obtained with coralline species also showed a decrease in total biomass, with similar results observed for various species of the same family tested in the wild (Villechanoux et al., Water 2022, 14(7), 1071; Koido et al., Zoological Studies 2022, 61:46).

[0015] Among the techniques recently adopted in the development of coral repopulation projects is the technique of "micro-fragmentation" or "reskinning" developed by David Vaughan for certain types of slow-growing massive corals that form ball-shaped or very large mounds, such as brain corals ("The culture of massive corals using "micro-fragmentation" for the "reskinning" of degraded coral reefs," Christopher Page and David Vaughan, conference paper, March 2014).

[0016] Such techniques, which utilize the homotypic fusion of coral fragments to accelerate coral growth rates (growth rates), have been shown to be applicable only to common corals. While this technique significantly reduces the time it takes for corals to cover a large area with a single thin layer, it does not affect the coral's three-dimensional structure, producing only thin attachment structures rather than ball- or mountain-shaped structures (Vaughan et al., 2014). Such techniques are not effective in promoting the three-dimensional growth of arborescent corals, such as those in the Coralaceae family.

[0017] For these reasons, there is a strong demand to develop economically sustainable and effective strategies that allow the cultivation of precious corals, which would guarantee new sources of raw material for the coral industry and are of great importance from an environmental point of view, in order to limit the overexploitation of wild populations of these species, which are already severely affected by economic overfishing. Summary of the Invention

[0018] Therefore, the technical problem underlying the present invention is to provide a method for advantageously promoting the production and / or cultivation of precious corals or colonies of precious corals, as well as their recruits or chimeras, from existing fragments, colonies, or recruits belonging to the Coralline family, which makes it possible to meet the above-mentioned needs, as stated with reference to the prior art.

[0019] The present invention is based on the discovery that fixing fragments or recruits of precious corals of the Corallinae family to a support element in a suitable arrangement, particularly by placing the fragments or recruits at an appropriate distance from each other, can promote fusion between adjacent fragments or recruits. This avoids the long growth period for precious coral colonies, recruits, or chimeras, and particularly significantly shortens the time required to form new colonies, recruits, or chimeras of precious corals of a desired size. Furthermore, the method of the present invention utilizes the ability of precious coral fragments or recruits to fuse with each other through allogamy, allogamy, interspecies fusion, or chimerism when fixed to a support element in a specific arrangement devised by the inventors.

[0020] In particular, the inventors have found that fusion between adjacent gem coral fragments or recruits can be achieved or promoted by placing adjacent fragments or recruits in close contact with each other or by arranging them so that the minimum distance between them is 2 cm or less, preferably 5 mm or less, and even more preferably 0 mm. Furthermore, the inventors have found that the basal diameter of the gem coral or the resulting colony can be maximized, thereby increasing its commercial value. Surprisingly, fusion between colonies of gem coral that are 2 to 4 years old, as well as fusion between gem coral fragments arranged according to the optimal arrangement identified by the inventors, is advantageous in significantly shortening the time required to produce gem corals of the desired size from decades to just a few months. This makes sustainable aquaculture possible on an industrial scale.

[0021] In one embodiment, the method of the present invention can provide new raw gem corals of significant commercial value. In particular, "x" number of gem corals (x=2, 3,..., n, each weighing m1, m2,..., m n ) fragments, each of which is placed in close contact with at least the adjacent fragment or within 2 cm of the adjacent fragment, and cultured under conditions suitable for their growth, can induce fusion between the adjacent fragments. This allows for a total mass of m T ≒m1+m2+...m n However, the total value v T >>v1+v2+...v n As is well known, the value per kilogram of raw gem coral actually increases sharply as the size of the coral increases, especially as the diameter of the base increases.

[0022] Known techniques in the field of culturing common coral fragments are not based on fragment fusion and require long waiting periods, i.e., long cultivation periods, to obtain corals of suitable size. The method of the present invention is different from this, and is advantageous because it allows the use of a large number of precious coral fragments without prolonging the coral formation period. In fact, the inventors have discovered that the period required for fusion between fragments is highly dependent on the distance between the fragments, but is less dependent on various biological and non-biological factors (e.g., species used, temperature, water flow, nutrient concentration, pH, etc.), and is not dependent on the number of fragments used.

[0023] Therefore, in the method of the present invention, by increasing the number of fragments, it may be possible to produce raw material corals with a larger diameter than the maximum diameter obtainable in nature without extending the production period.

[0024] The geometry employed by the method of the present invention is effective in manipulating the basal diameter of precious corals and promoting their formation and three-dimensional growth, and represents a surprising novelty compared to the results observed with micro-fragmentation techniques, which are only effective at a two-dimensional level and only form thin layers of coral.

[0025] The technological solution used to fix the coral fragments to the support is innovative compared to the solutions used in micro-fragmentation techniques, allowing the diameter of the base of the precious coral to be manipulated.

[0026] In fact, placing fragments of massive common corals less than 2 cm apart during micro-fragmentation has not been shown to promote rapid, widespread recovery. This is because the growth rate of massive common corals declines rapidly once fusion between micro-fragments begins, rendering the micro-fragmentation technique ineffective if fusion occurs prematurely (Vaughan et al., 2015).

[0027] Conversely, the inventors have found that using an arrangement in which the fragments are fixed to a single support element with an inter-fragment distance of less than 2 cm, preferably less than 5 mm, and more preferably 0 mm, actually promotes growth in trunk diameter of coral colonies in a much shorter period of time than would be required in nature.

[0028] Furthermore, the new arrangement for precious corals developed by the inventors has significant advantages over the micro-fragmentation technique for massive corals. In fact, in the first qualitative experiment using red coral, it was surprising to find that the growth rate of the fragments did not drop sharply even after fusion began, unlike in the case of micro-fragmentation of massive corals. On the contrary, the fused fragments showed a higher growth rate and a higher survival rate than the reference single fragments.

[0029] This unexpected effect allows colonies generated by the method of the present invention to tolerate transplantation better and grow faster than single fragments.

[0030] This is a very important aspect in the culture of corals, especially those belonging to the Corallinae family, as it allows overcoming the main technical challenges that make the culture of these corals so difficult: long culture periods in nurseries and biomass loss during transplantation.

[0031] Without wishing to be bound by theory, it is hypothesized that this occurs because fused coral fragments are larger in size than single fragments and have a greater number of initial polyps. In fact, for corals of the same age, larger corals generally have higher growth and survival rates, produce far more eggs and sperm sacs per polyp, and are more reproductive. They also have better resilience and regeneration abilities, and a greater photosynthetic capacity. They also have more available energy because they are exposed to more water and can capture more prey.

[0032] Another important advantage of the method of the present invention is that, unlike the techniques currently used in the cultivation of slow-growing corals such as massive corals, it is effective in shortening the cultivation period of precious corals, regardless of the fusion mechanism selected between fragments or recruits.

[0033] In fact, it is now believed that the only fusion mechanism available for practical purposes in coral culture is represented by allogamy, which requires the use of only corals of the same "parent" origin, since fusion between different genotypes (i.e., heterologous species) inevitably results in tissue rejection, dominance of one genotype, and often the death of the recessive genotype.

[0034] Conversely, the method of the present invention offers the possibility of using equivalent genotypes or different species to create gem corals with colors and large color contrasts that are very rare or nearly non-existent in nature.

[0035] In particular, the inventor has discovered that when placing a single recruit of precious coral that is less than 6 months old and that has been attached or added to a substrate suitable for growth on a single support element, by placing it in close contact with adjacent recruits or by setting the minimum distance between adjacent recruits to 5 mm or less, and more preferably 0 mm, the selected adjacent recruits will fuse together, making it possible to form a new chimera of precious coral.

[0036] Advantageously, placing a single recruit of precious corals less than six months old according to the distance and placement according to the method of the present invention can be used to expand the resulting colony and furthermore to be used in subsequent culture cycles according to the method of the present invention. This procedure can increase the productivity of the seed bed. Indeed, increasing the number of individuals per colony from 1 to n individuals increases the colony's biomineralization (growth rate) and the amount of food captured (energy) by approximately n-fold.

[0037] The method of the present invention is particularly versatile and can be easily implemented in open water or in controlled environmental areas such as tanks or aquaria using a wide variety of solutions and equipment suitable for cultivating and growing corals known in the art.

[0038] The present invention ensures the production of precious coral colonies of a desired size in a shorter time than methods known in the art, and is particularly capable of producing colonies that are tolerant to transplantation and have a higher reproductive potential. Currently, the cultivation of Coralline precious coral species is significantly limited by numerous technical difficulties. Therefore, the method of the present invention has great potential for the development of active repopulation projects for Coralline precious coral colonies.

[0039] For example, the method of the present invention can suitably modify conventional coral cultivation techniques that have not been effectively applied to Corallinae until now by performing a preliminary step of cultivating precious coral fragments in a land-based fusion nursery, allowing fusion between the fragments to occur before the conventional nursery culture step, followed by a step of transplanting them to a re-propagation site. The preliminary step in the fusion nursery shortens the long period required for the fragments to reach an ideal size suitable for transplantation, thereby shortening the culture period in the propagation nursery and enabling the production of colonies that are more tolerant to transplantation.

[0040] Furthermore, because large colonies have a high reproductive capacity, the fused corals obtained by the method of the present invention can serve as breeding bases, enhancing the natural recovery of coral populations that are currently being overfished.

[0041] The method for cultivating precious corals by fusion devised by the inventor is particularly useful in the jewelry industry, which is currently facing a crisis due to a shortage of raw materials. Advantageously, the method of the present invention can actually be used to produce corals and precious stones in shapes and sizes that could not be realized with so-called natural corals.

[0042] Another application of the method according to the present invention is particularly relevant to the tourism sector, which has been actively involved for many years in the development of active repopulation projects for precious coral colonies. For example, resort owners with access to the sea, especially along the Mediterranean coast, could be offered the opportunity to implement active restoration programs for precious corals based on the method according to the present invention. These programs could include, for example, the installation of energized metal structures (in-situ nurseries) in front of beaches to improve the snorkeling experience, and the installation of land-based propagation nurseries to display live corals to non-swimmers.

[0043] The benefits of this type of partnership are mutual, indeed, on the one hand it strengthens the efforts of environmental organisations to involve private actors in conservation activities, and on the other hand it benefits tourism operators, not only in terms of their image but also as various studies have shown that operators on the ground are aware of the potential increased economic benefits that can come from having usable reefs, natural barrier reefs, reefs, artificial barrier reefs and electric reefs near their beaches.

[0044] Thus, the present invention relates to a method for producing or cultivating precious corals, or precious coral colonies, recruits, or chimeras, comprising the steps of: a) Prepare at least two fragments or recruits of existing precious corals belonging to the Corallinae family. b) fixing said fragments or said accessions to at least one support element so that the minimum distance between each said fragment or said accession and at least one other said fragment or said accession is 0 cm to 2 cm, preferably 0 mm to 5 mm, more preferably 0 mm, 1 mm, 2 mm, or 3 mm; c) The fragments or accessions fixed in step b) are cultured in seawater at temperatures ranging from 2°C to 30°C.

[0045] Other important features of the method according to the invention are defined in the dependent claims. Further advantages and features of the invention will become apparent from the following detailed description.

[0046] Glossary Terms used herein are as commonly understood by those of ordinary skill in the art unless specifically stated otherwise.

[0047] Within the scope of the present invention, the term "precious coral" refers to a coral or coral colony belonging to the Corallinae family that is not a reef-forming coral, does not possess zooxanthellae, and can survive at relatively deep water depths because it feeds on plankton or obtains nutrients through osmosis. The skeleton of a precious coral is composed of a complex of calcium carbonate and protein. Generally, precious corals are dendritic, long-lived organisms with low growth and reproduction rates. In any respect in this specification and claims, the expression "precious coral belonging to the Corallinae family" can be substituted with the expression "colony of precious coral belonging to the Corallinae family."

[0048] In the context of the present invention, the expression "Corallinae" refers to a family of corals of the phylum Cnidaria, class Anthozoa, subclass Octocorallia, order Alcyonacea and suborder Scleractinia.

[0049] In the present invention, the term "red coral" is used as a synonym for the precious coral species Corallium rubrum or its colonies. In the present invention, red coral is a species of Octocorallia belonging to the family Corallinae (genus Corallium) and is widely distributed in the Mediterranean and eastern Atlantic. Red coral is the only coral species found in the Mediterranean Sea and is distributed in areas from Greece to Tunisia, including the Strait of Gibraltar, Corsica, Sardinia, Sicily, and the Balearic Islands. However, it is also widely distributed in the eastern Atlantic, including Portugal, the Canary Islands, Morocco, and the Cape Verde Islands, where it is found at depths of up to 200 meters, usually in areas with low light and sparse vegetation. Red coral prefers shady and enclosed areas (such as semi-dark caves, rock overhangs, and crevices) and can live at depths of 20-30 meters to 200 meters. Red coral colonies have been found at depths of 600-700 m in the Strait of Sicily and at depths of up to 1061 m in the waters of the Maltese Islands (Knittweis L. et al., 2016, 41st CIESM General Assembly), but the size of deep-sea red coral populations remains poorly understood.

[0050] Red corals are a very slow-growing species. Growth rate studies have shown that colonies grow an average of 0.25 to 0.66 mm in basal diameter per year. Scientific literature suggests that it takes red corals approximately 30 to 35 years to reach a basal diameter of 7 mm, and 75 to 100 years to reach a commercially valuable diameter.

[0051] Red corals form branching colonies that can grow to over 20-30 cm in height and are generally bright red in color, although they can also be pink. Completely white albino colonies are rarely observed. Red coral polyps are white and transparent, with feathery tentacle edges that can be seen when they extend their tentacles to capture prey.

[0052] In the context of this specification, the terms "coral nursery," "coral rearing facility," "fusion nursery," and "propagation nursery" are used synonymously and refer to any device known in the art suitable for cultivating corals in a controlled environment, such as in saltwater, underwater, or in natural or artificial reservoirs, tanks, aquaria, etc.

[0053] In the context of this specification, the term "thermocline" refers to the transition zone or layer between the surface mixed layer and the deep water layer in deep water bodies such as oceans or seas. The thermocline experiences a steep temperature drop over a range of several meters from the water temperature values ​​expected in the surface mixed layer to values ​​corresponding to the temperature of deep water, with a gradient of up to 10 degrees. The temperature of deep water remains relatively constant, usually below 20 degrees, even in summer.

[0054] In the context of the present invention, the expression "fragment of an existing gem coral colony" or "fragment of an existing gem coral" refers to (i) a fragment or part of a living gem coral colony, or (ii) a living gem coral colony, particularly a living gem coral colony that is between 6 months and 4 years old, preferably between 2 and 4 years old.

[0055] In any respect in this specification and claims, the expressions "fragments of precious coral colonies" or "fragments of precious coral" may be replaced with the following expressions: (i) A part of a precious coral or a part of a precious coral colony, in particular a part of a precious coral or a part of an adult precious coral colony, more in particular a tip obtained from a branch of a precious coral or a precious coral colony; or (ii) Precious coral colonies, particularly those aged between 6 months and 4 years, preferably between 2 and 4 years.

[0056] In a particular embodiment, as will be explained in more detail below, the precious coral colonies (ii) that can be used in the method according to any one of the variations of the present invention are small-scale, where "small-scale" refers to those having an average length or diameter of 0 mm to 50 mm and / or an average thickness of 0 mm to 5 mm.

[0057] At all points in this specification and claims, the term "adult" may be substituted for the phrase "over 4 years of age."

[0058] At any point in this specification and the claims, the plural form "of precious corals" may be replaced with the singular form "of precious coral."

[0059] In the context of the present invention, with respect to a fragment of a precious coral or a fragment of a precious coral colony as a part or tip of a precious coral or colony thereof, the term "length" refers to the extension along the maximum size of such a fragment, and in particular the extension along the longitudinal growth of such a fragment. In particular, the term "length" refers to the distance between the oldest and youngest ends of the fragment in question.

[0060] In the context of this invention, with respect to a precious coral colony, fragment, recruit, or chimera, the term "width" refers to the size of the cross section of the base of the coral in any direction through the center or axis. In the context of this invention, the terms "cross section," "width," and "diameter" are synonymous.

[0061] With respect to disc-shaped or approximately disc-shaped gem coral colonies according to any one of the variations described in this specification and claims, the term "thickness" can be replaced with the term "height," and preferably refers to the thickness, height, or length measured at the center of the subject colony along its biomineralization axis. In the context of the present invention, the terms "thickness," "height," and "length" are synonymous.

[0062] In the context of the present invention, the terms "precious coral recruit," "coral recruit," and "recruit" are used interchangeably and refer to one or more specimens or individuals of precious coral belonging to any one of the species described herein that have completed the process of attachment (i.e., settlement) or attachment to a suitable attachment or growth substrate but have not yet begun, or are about to begin, or have already begun the process of metamorphosis, as well as colonies that have been attached or attached to a substrate by such attachment and / or metamorphosis processes and are between 0 and 6 months old.

[0063] As known to those skilled in the art, the process by which precious coral larvae or planulae attach to a suitable attachment or growth substrate is defined in the art as "coral recruitment." During the first few weeks after settlement or attachment to the attachment substrate, the attached precious coral larvae undergo metamorphosis into polyps and then begin to expand their tissues onto the substrate to enhance attachment.

[0064] In the context of the present invention, the term "chimera" refers to a colony resulting from the fusion of two or more precious coral recruits with different genomes, attached or affixed to a suitable attachment or growth substrate, preferably in contact with each other, according to any of the embodiments described herein, particularly before the recruits have developed a self-recognition system (immune system) capable of rejecting tissue with different genomes.

[0065] In one embodiment of the present invention, when precious coral fragments or accessions according to any one of the variations described herein are fixed to at least one support element via one end or part thereof, as described in detail below, the "minimum distance" between the fragments or accessions refers to the distance between the bases of the fragments or accessions that are supported by or fixed to the support element.

[0066] In the context of the present invention and claims, the terms "attachment substrate," "growth substrate," and "substrate suitable for growth" are used interchangeably and refer to substrates known to those skilled in the art that allow attachment (i.e., settlement) of precious coral larvae and facilitate the subsequent metamorphosis and growth process to form precious coral recruits. These substrates include any type of seabed suitable for coral settlement and growth.

[0067] At any point in this specification and claims, the word "comprising" may be substituted with "consisting of." DETAILED DESCRIPTION OF THE INVENTION

[0068] As mentioned above, the present invention relates first to a method for producing or cultivating precious corals, or colonies, recruits, or chimeras of precious corals, comprising the steps of: a) Prepare at least two fragments or recruits of existing precious corals belonging to the Corallinae family. b) Fixing said fragments or said accessions to at least one support element so that the minimum distance between each of said fragments or said accessions and at least one other of said fragments or said accessions is 0 cm to 2 cm, preferably 0 mm to 5 mm, more preferably 0 mm, 1 mm, 2 mm, or 3 mm. c) The fragments or the accessions fixed in step b) are cultured in seawater at a temperature ranging from 2°C to 30°C, preferably from 16°C to 22°C.

[0069] The method of the present invention, according to any one of the embodiments set forth in the specification and claims, makes it possible to produce or cultivate new precious corals, or precious coral colonies, recruits, or chimeras, from precious coral fragments, existing colonies, or recruits.

[0070] In one aspect, the method of the present invention according to any one of the embodiments set forth in the specification and claims makes it possible to generate or cultivate a new precious coral or a colony thereof from at least two fragments of an existing precious coral belonging to the Corallinaceae family.

[0071] In a further aspect, the method of the present invention according to any one of the embodiments set forth herein and in the claims allows for the generation or cultivation of a new colony of precious coral from at least two recruits of an existing precious coral belonging to the Corallinaceae family.

[0072] In a further aspect, the method of the present invention according to any one of the embodiments set forth herein and in the claims allows for the generation or cultivation of new chimeras of precious corals from at least two recruits of existing precious corals belonging to the Corallinaceae family.

[0073] In certain aspects, the method of the present invention according to any one of the embodiments set forth herein and in the claims allows for the production of new precious corals or precious coral colonies of predetermined sizes from fragments, existing colonies, or recruits of precious corals belonging to the Coralline family, in a significantly shorter time than would be required to obtain the same sizes of precious corals, precious coral colonies, recruits, or chimeras in nature from fragments, existing colonies, or recruits of precious corals belonging to the Coralline family.

[0074] In fact, as mentioned above, by fixing each precious coral fragment or recruit so that it is in close contact with at least one other such fragment or recruit, or at a minimum distance of 2 cm, it is possible to promote fusion between adjacent fragments or recruits, which makes it possible to form large precious corals or colonies in a much shorter time than the time it takes to obtain corals or colonies of the same size from a single fragment or single recruit of the target species, as reported in the literature.

[0075] In particular, the method of the present invention in any one of the variations shown in this specification makes it possible to produce new colonies of precious corals with a base diameter, i.e., trunk diameter, that is much larger than that expected of a colony of the same height, degree of branching, and age in nature.

[0076] According to a preferred aspect, the method of the present invention according to any one of the embodiments set forth herein and in the claims can produce new precious corals or colonies of a predetermined size in 2 years or less, more preferably 1 year or less, and even more preferably 12 to 24 months, 8 to 12 months, or 4 to 8 months, which periods may vary within the ranges stated herein depending on the mutual distance between the fragments fixed to the support element.

[0077] According to a further aspect, the method of the present invention allows the generation of new colonies of precious corals from existing recruits of precious corals belonging to the Corallinae family in 4 years or less, or 2 years or less, more preferably 1 year or less, and even more preferably 12 to 24 months, which periods may vary within the ranges stated herein depending on the mutual distance between the individuals fixed to the support element.

[0078] According to one aspect, the fragments or recruits selected for use in the method of any one of the embodiments described herein belong to or are obtained from the same species of precious coral, the same colony of precious coral, or a different colony of the same species of pre-existing precious coral.

[0079] According to a further aspect, the fragments or accessions selected for use in the methods according to any one of the embodiments described herein belong to or are obtained from different species of precious coral or from different existing colonies of different species of precious coral.

[0080] According to a further aspect of the invention, the at least two fragments or the at least two recruits belong to or are obtained from the same species of precious coral belonging to the genus Coral or Pleurocorallium, in particular one of the red or pink corals of the genus Pleurocoral.

[0081] In a further embodiment, said at least two fragments or said at least two recruits belong to or are obtained from different species of precious coral belonging to the genus Coral or Pleurocorallium, in particular a different species of red coral or pink coral of the genus Coral.

[0082] In a preferred embodiment, the at least two fragments or the at least two accessions belong to one or more species of precious coral selected from Corallium rubrum, Corallium japonicum, Pleurocorallium elatius, Pleurocorallium konjoi, Pleurocorallium secundum, Hemicorallium regale, Hemicorallium laauense, and Hemicorallium sulcatum, or are obtained from one or more colonies of precious corals selected from these species. According to a preferred embodiment of the method of the present invention, the fragments or accessions are obtained from one or more colonies of coral belonging to one or more species and selected from the species set out in Table 1 below. [Table 1]

[0083] In a preferred embodiment of the present invention, the at least two fragments or at least two recruits are fragments of red coral recruits or are obtained from one or more colonies of red coral.

[0084] In a particular aspect, the method of the present invention according to any one of the embodiments described herein can produce chimeras of precious corals from at least two recruits belonging to the same or different species of precious corals belonging to the Corallinaceae family, particularly the genus Corallinus or Pleurocorallium.

[0085] The following are examples of further characteristics of precious coral fragments and recruits that can be used in the methods according to any one of the embodiments described herein. Fragments of precious coral or fragments of precious coral colonies

[0086] According to one aspect of the present invention, the fragments suitable for use in the methods of the present invention are apical portions of precious corals obtained from branches of colonies according to any one of the embodiments described herein, i.e., the tops or "tips" of precious coral branches.

[0087] The "tips," or tops, of precious corals are generally too thin to be processed into gemstones and have little commercial value in this sector, as most are treated as waste. This makes the use of these fragments particularly advantageous. In fact, the Mediterranean General Fisheries Commission recommends, although not obligatory, that these parts be discarded on-site directly from the vessel during fishing. The use of these fragments avoids the need to harvest parts of precious corals found in nature.

[0088] According to one aspect of the present invention, the apical end of the precious coral used in the method according to any one of the embodiments described herein has a generally cylindrical shape, although apical ends of precious corals having any shape can be used in the method according to any one of the embodiments described herein.

[0089] According to one aspect of the present invention, the fragments used in the methods of the present invention are harvested from precious corals or donor colonies of precious corals, preferably adults. Alternatively, the fragments may be obtained from fragments of such colonies previously grown in in situ and / or ex situ coral propagation nurseries. The length of the fragments that can be used in the methods according to any one of the embodiments described herein is preferably at least 0 cm or 0.5 cm.

[0090] According to a preferred aspect of the method according to any one of the embodiments described herein, the length of each selected fragment is between 2.5 cm and 5 cm, preferably between 1.5 cm and 3.5 cm, more preferably 2.5 cm.

[0091] As mentioned above, the pieces of precious coral that can be used in the method according to any one of the embodiments described herein may be portions of precious coral of various shapes, and are preferably portions of roughly cylindrical precious coral.

[0092] Fragments suitable for use in the method according to any one of the embodiments described herein are in particular portions of precious coral of any shape or diameter. In a preferred embodiment of the method according to the present invention, the fragments are top fragments having a length of 2.5 cm to 5 cm and a cross-sectional or average diameter of 2 mm.

[0093] According to one aspect of the present invention, step a) of the method according to any one of the embodiments described herein comprises cutting or severing the fragments from one or more colonies of precious coral belonging to the Coralline family selected according to any one of the variations described herein.

[0094] Fragments can be cut or trimmed from one or more of the selected source colonies using any one of the techniques known to those skilled in the art. For example, they can be trimmed using scissors or any cutting pliers conventionally used in the art for this purpose (e.g., wire cutters), provided that they have a fine, sharp blade. To expedite collection of fragments in the ocean, selected colonies can be fragmented using a coral pick or hammer. The resulting fragments can be collected and trimmed on land using scissors or other cutting means.

[0095] According to a further aspect of the present invention, the fragments suitable for use in the method of the present invention are precious coral colonies, in particular colonies of precious coral that are between 6 months and 4 years old, preferably between 2 and 4 years old.

[0096] Precious coral colonies suitable for use in the method according to any one of the embodiments described herein are those that are of sufficient size to allow detachment from an attachment or growth substrate and then fixation onto one of the support elements according to any one of the forms shown herein, particularly colonies of precious coral between six months and four years old.

[0097] Based on the species and age of the target precious coral, one skilled in the art can easily identify precious coral colonies having a size that allows them to be easily and safely detached from the attachment or growth substrate without damaging its vitality, and then fix them to at least one support element according to any one of the forms exemplified herein. Furthermore, one skilled in the art can detach colonies of a selected age from the attachment or growth substrate in a delicate and precise manner that does not damage the coral.

[0098] In a preferred embodiment, the age of the fragments as precious coral colonies is between 2 and 4 years. As known to those skilled in the art, precious coral colonies that are between 2 and 4 years old are often optimally sized to allow for detachment from the attachment or growth substrate and subsequent fixation onto a support element of any one of the forms shown herein.

[0099] In one embodiment of the present invention, the fragments as a colony of precious coral have an average width or diameter of 0.5 cm to 5 cm, and / or an average thickness or length of 2 mm to 5 mm.

[0100] Colonies of the above sizes can be, for example, asexually fragmented from a larger coral colony. Alternatively, precious coral colony fragments suitable for use in the methods of the present invention can be generated from at least two recruits of precious coral using the method of any one of the embodiments described herein, particularly the method described in Example 3.

[0101] In one aspect of the present invention, the fragment as a colony of precious coral is a precious coral chimera obtained or produced from at least two precious coral recruits by using a method according to any one of the embodiments described herein.

[0102] In a particularly preferred embodiment, the shape of the colony of precious coral according to any one of the variations described herein is disc-shaped or approximately disc-shaped, and particularly preferably, its average height or thickness is at least 2 mm, at least 3 mm, or at least 5 mm, and its average diameter is 5 mm to 15 mm.

[0103] Indeed, it is known to those skilled in the art that when precious coral recruits grow into colonies that are disc-shaped or approximately disc-shaped with an average thickness of about 2 mm to 5 mm, such colonies can be more easily removed from the attachment substrate without causing significant damage and can then be fixed to at least one support element according to any one of the variations shown in this specification.

[0104] In a more specific embodiment, the said colony of precious coral is a colony of red coral that is 2 to 4 years old, disc-shaped or approximately disc-shaped, 10 mm to 15 mm in diameter, and 2 mm to 5 mm in height or thickness.

[0105] Colony fragments aged 6 months to 4 years according to any one of the variations described herein may be collected directly from the selected area or may be generated from a single recruit of a precious coral previously grown in an in situ and / or ex situ coral propagation nursery.

[0106] In one aspect, these colonies can be produced from a single accession of precious coral attached or affixed to a suitable attachment or growth substrate by carrying out a method according to any one of the embodiments described herein. Specifically, each of the colonies can be obtained by fixing two or more accessions attached to a particularly suitable attachment or growth substrate on a suitable support element such that the minimum distance between each accession and at least one other accession is 0 mm to 5 mm, more preferably 0 mm to 3 mm, and even more preferably 0 mm to 1 mm.

[0107] Alternatively, precious coral colonies that can be used in the methods of the present invention can be obtained from precious coral recruits using any conventional method known to those skilled in the art, such as by recruiting one or more precious coral larvae onto a suitable attachment or growth substrate and culturing the recruits thus obtained for the required period of time under optimal growth conditions.

[0108] As will be discussed in more detail below, precious coral larvae can be recruited directly in marine settings or artificial tanks by contacting them with a substrate capable of promoting larval attachment (or settlement). Examples of attachment or growth substrates suitable for use in capturing precious coral larvae include polyvinyl chloride (PVC), steel, glass, marble, or other materials to which larvae can be attached or attached, preferably without macroporosity that may make subsequent removal of the colony difficult. By way of example only, marble tiles of 30 mm x 30 mm or other sizes could also be used. The process of larval settlement on the substrate is stochastic, and the number of larvae captured is proportional to the surface area available for settlement. For this reason, it is preferable for the support to have as large a surface area as possible without complicating anchoring to the seafloor.

[0109] When the juveniles are captured in tanks, the size of the support used is preferably a submultiple of the tank bottom so that the entire bottom of the tank is covered. A person skilled in the art can determine the size of the support based on the method of anchoring to the seabed used or based on the size of the tank. Precious coral recruits

[0110] As noted above, according to the present invention, precious coral recruits suitable for use in the methods of any one of the embodiments described herein include or are represented by one or more individuals or specimens of precious coral of any one of the species described herein that have completed the process of attachment (i.e., settlement) to a suitable attachment or growth substrate but have not yet begun, or are about to begin, or have already begun the process of metamorphosis, as well as colonies that have attached or affixed to the substrate by such attachment and / or metamorphosis processes, and are 6 months of age or less, preferably 3 months of age or less, and more preferably 1 month of age or less.

[0111] In certain embodiments of the present invention, the recruits each comprise at least a larva of a precious coral according to any one of the species described herein, which has just completed the process of attachment or attachment to a suitable attachment or growth substrate and has not yet begun or completed the process of metamorphosis into a polyp.

[0112] In further embodiments, the recruits each comprise at least a precious coral larva from any one of the species described herein, which has been attached or affixed to the substrate and has just completed the process of metamorphosis into a polyp.

[0113] In further embodiments, the recruits each include at least colonies less than six months old that result from one or more precious coral larvae attaching (or settling) to a suitable attachment or growth substrate, metamorphosing and growing on said substrate.

[0114] As mentioned above, the recruits can be obtained from the settlement of larvae of the same or different species of precious coral according to any one of the variations described herein.

[0115] In one embodiment, the recruits can be obtained from the capture of at least two larvae of precious corals, which can be done according to any one of the methods known to those skilled in the art, such as those previously described herein.

[0116] For example, larval recruitment can be achieved near a breeding population of a selected coral species, e.g., red coral, by securing a suitable attachment substrate to the seafloor via a waterproof adhesive, such as a two-component resin or any other adhesive suitable for use in water. Alternatively, larval capture can be achieved by tethering the selected attachment substrate to the seafloor with a mechanical anchor or by tethering it to biological remains heavy enough to sink. A typical weight for biological remains is approximately 5 kg, but this can be selected by those skilled in the art based on the characteristics of the selected site, such as water currents and the type of seabed. It is important to lower the center of gravity and concentrate the weight downward to prevent the biological remains from tipping over during sinking.

[0117] In a further embodiment, larvae can be harvested by introducing at least an attachment substrate into a tank or facility suitable for growth and / or maintenance containing selected precious coral larvae. This allows the larvae to attach or settle on the substrate. This procedure is particularly advantageous because it avoids larval dispersal and allows for more detailed characterization of the larvae's genome. Furthermore, the high density of larvae in the tank increases the number of recruits per unit of surface area. Those skilled in the art can select a facility more suitable for cultivating and maintaining a selected species of precious coral and can adjust the amount and frequency of coral feeding based on the available facility tanks. By way of example only, red coral propagation colonies can be harvested before the breeding season and cultured in a tank containing a support suitable for larval production and settlement. The most commonly used solution for red coral maintenance is represented by a dual-circuit system, which operates as a closed circuit for 90 days after larval production (i.e., spawning) and then as an open circuit thereafter. The colony can be fed with Artemis larvae, homogenized algae, and homogenized fish and crustaceans, while the circuit can be supplied with filtered seawater chilled at 12 to 16°C.

[0118] In a further embodiment, the capture of larvae can be carried out in the ocean using one or more suitable nets in a process for producing wild precious coral larvae. The larvae thus captured can be transferred to a suitable holding tank and exposed to an attachment substrate according to any one of the variations exemplified herein.

[0119] Alternatively, other methods, tools and materials known in the art can be used, such as marble tiles with a hole in the middle. For anchoring, holes are drilled into the seabed using an oil drill or other suitable tool depending on the type of seabed, and the bars are then inserted into the holes and secured with two-part epoxy or other fastening methods known in the art. The tiles are inserted into the holes and secured to the bars, in this case with Parker screws or two-part epoxy.

[0120] In certain embodiments, each of the at least two accessions of precious coral used in the methods of the present invention is attached, immobilized, or affixed to an attachment or growth substrate, which is a suitable substrate for attachment and growth of the accessions.

[0121] In other words, in certain embodiments, the accessions used in the methods of the present invention are individually attached or attached to their respective substrates suitable for their growth. In step b) of the methods of the present invention, the accessions can be fixed to a support element so that the minimum distance between at least one accession attached, attached, or immobilized on a substrate and at least one accession attached or attached on a different substrate is less than 2 cm, preferably less than 5 mm.

[0122] One aspect of the present invention relates to a method for producing or culturing a precious coral, or a colony, recruit, or chimera of a precious coral, from an existing recruit of a precious coral belonging to the Corallinaceae family, particularly according to any one of the variations described herein, the method comprising the steps of: a') Providing at least two recruits of an existing precious coral of the Corallinae family according to any one of the variations described herein, each of the recruits being attached or affixed to an attachment or growth substrate. b') Fixing each of the attachment or growth substrates to at least one support element so that the minimum distance between each accession attached or affixed to each of the substrates, or at least one of the accessions, and at least one of the accessions attached to another substrate is 0 mm to 5 mm. c) The substrate fixed in step b') is incubated in seawater at a temperature ranging from 2°C to 30°C, preferably from 16°C to 22°C.

[0123] The accessions according to any one of the variations described herein, each attached or affixed to a suitable attachment or growth substrate, can be obtained or generated by exposing one or more precious coral larvae to a suitable attachment or growth substrate according to any one of the variations described herein. In some cases, the accessions are cultured for a desired period of time. Finally, the selected attachment or growth substrate is cut to obtain a plurality of separate substrates, each with at least one accession, preferably one accession, attached or affixed thereto.

[0124] Preferably, the attachment or growth substrate to which the accessions usable in the methods according to any one of the embodiments described herein and claimed are attached or affixed comprises or consists of a material that can be cut with millimeter or sub-millimeter precision.

[0125] In particular embodiments, the attachment or growth substrate comprises or consists of a plastic material, in particular a polymeric material, hi particularly preferred embodiments, the attachment or growth substrate comprises or consists of a thermoplastic polymer, preferably polyvinyl chloride (PVC).

[0126] A further aspect of the present invention then relates to a method for producing or cultivating a precious coral, or a colony, recruit or chimera of a precious coral, from an existing recruit of a precious coral belonging to the Coralline family according to any one of the variations described herein, the method comprising the steps of: a') attaching or adding at least two larvae of the existing precious coral belonging to the Coralline family according to any one of the variations described herein to an attachment substrate or growth substrate according to any one of the variations described herein so as to obtain the at least two recruits. b') cutting the attachment or growth substrate to obtain a plurality of separate substrates with one of the accessions, preferably one of the accessions, attached or attached to each of them; b'') each of the plurality of substrates is fixed to at least one support element such that the minimum distance between at least one, preferably each, of the added particles attached or affixed to each of the plurality of substrates and at least one added particle attached to another of the plurality of substrates is preferably between 0 mm and 5 mm; c) The substrates immobilized in step b'') are cultured in seawater at a temperature ranging from 2°C to 30°C, preferably from 16°C to 22°C.

[0127] In a preferred embodiment, step b') is carried out such that one individual is attached or added to each of a plurality of separate substrates.

[0128] The cutting of the attachment or growth substrate is preferably carried out in such a way as to obtain a plurality of separate substrates of the same or different shape, thickness or size, provided that each substrate contains at least one of said accessions, preferably one of said accessions.

[0129] In particular, the cutting of the attached substrate is preferably carried out in such a way as to obtain a plurality of separate substrates, each of which comprises at least one of the added entities at a suitable distance from at least one of the peripheries of the substrate thus cut. "A suitable distance from at least one of the edges" particularly means a distance at which the added entities attached or added to adjacent substrates fixed in contact with each other are spaced apart from each other at a minimum distance of preferably 0 to 5 mm when each substrate is fixed to a support element according to any one of the variants described herein.

[0130] In other words, a suitable distance from the periphery can be determined so that when each substrate is secured to the support element, the edges of the substrates are in close contact with each other and the individual members on each substrate are at a selected distance from each other, thereby avoiding gaps between the substrates after securing.

[0131] To avoid steps or height differences between the separated substrates, it is preferable to make the substrates all the same thickness and then fix them to the support element so that at least one edge is in contact with the edge of another substrate.

[0132] In a preferred embodiment, said step b'') is then carried out such that at least the periphery of each of said substrates is in contact with the periphery of at least one other substrate of said substrates.

[0133] The size of each of the substrates is preferably such that an inductee attached or added to each substrate has enough space to grow to a diameter of 1.5 cm without reaching an edge that is not in contact with another substrate.

[0134] One skilled in the art can determine the size of the substrate so that, after being secured to a support element, at least one of the members on that support element will be positioned at a desired distance from at least the other members and will have enough space to grow without reaching edges that are not in contact with other substrates, as will be explained in more detail below.

[0135] If the edges of the substrates that come into contact with one another are not precise and gaps remain due to cutting irregularities, the adhesive or other suitable material used to secure the substrates to the support element can be used to fill in the gaps between the substrates.

[0136] Before carrying out step b) of the method according to any one of the embodiments of the present invention, the substrate to which at least the precious coral fragments, recruits, or precious coral recruits according to any one of the variations described herein are attached, affixed, or immobilized is preferably stored in seawater at a temperature range of 16°C to 22°C and is not exposed to direct sunlight.

[0137] For ease of further manipulation, fragments or recruits can be separated based on species or locality of the colony, for example, into separate containers which can then be combined into a larger container suitably filled with chilled seawater.

[0138] All equipment used for cutting, preparing, or storing precious coral fragments or recruits for use in the methods of the present invention should preferably be properly sterilized or disinfected, for example with 70% ethanol, and rinsed with freshwater to avoid contamination of the fragments or selected recruits with undesirable microorganisms. It is also recommended that sterile surgical gloves be used when handling precious coral fragments or recruits during any one of the steps of the methods of the present invention.

[0139] According to one aspect of the present invention, step a) of the method according to any one of the embodiments described herein involves providing a plurality of fragments of recruits of a colony of precious coral belonging to the Corallinaceae family, or a plurality of attachment substrates to which at least recruits of precious coral according to any one of the variations described herein are attached, affixed or immobilized.

[0140] In a further aspect, step a) of the method according to any one of the embodiments described herein comprises providing at least two fragments and at least two recruits of a colony of a precious coral belonging to the Corallinaceae family according to any one of the preceding variations.

[0141] The number of fragments, accessions, or substrates selected for use in the method according to any one of the embodiments described herein is preferably between 2 and 60, more preferably at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 25, at least 30.

[0142] Those skilled in the art will be able to select a more appropriate number of fragments or recruits based on the desired size of the precious coral, or its colony or recruit, obtained by the method of the present invention, i.e., the size of the coral, or its colony or recruit, obtained by fusion of the fragments or recruits used in the method of the present invention.

[0143] For roughly cylindrical fragments, if many fragments are used to achieve a size larger than that obtainable in nature, multiple fusions can be performed to avoid excessive tissue necrosis in the central fragment and minimize the possibility of abnormal skeletal density in the final colony. For example, four fragments can be first secured to the vertices of a square, then fused to form an octagon by placing four additional fragments at the center of each side of the square, then eight additional fragments at the center of each side of the octagon, adding another 16 fragments, and so on.

[0144] Prior to carrying out fixation step b) according to the method of the present invention, the selected fragments or recruits are preferably visually inspected, for example to assess the presence of pathologies and possible tissue necrosis, and, if possible, to check the vitality of the polyps. Such inspection is useful for discarding fragments or recruits that are not viable or do not meet the desired requirements.

[0145] As mentioned above, in step b) of the method according to any one of the embodiments of the present invention, the fragments or recruits prepared in step a), or the attachment substrate to which at least one of the above recruits is attached, added or immobilized, respectively, are fixed to at least one support element in a precise arrangement suitable for promoting fusion between adjacent fragments or recruits, which then results in the formation of a precious coral, its colony or recruit having a desired size.

[0146] Fusion between adjacent fragments or adjacent recruits spaced at a minimum distance as defined in any one of the limitations defined herein and in the claims can occur by different mechanisms, such as, for example, homotypic fusion, heterotypic fusion, interspecies fusion and / or chimerism, depending on the type of fragment or recruit selected.

[0147] In particular, fusion between adjacent recruits spaced at a minimum distance as defined in any one of the limitations set forth in this specification and claims can occur by chimerism, resulting in the formation of one or more chimeras of precious corals.

[0148] As mentioned above, in order to promote fusion between immobilized fragments or recruits, it is important that the fragments or recruits are immobilized so that the minimum distance between each fragment or recruit and at least one other fragment or recruit is 2 cm or less, preferably 1 cm or less.

[0149] In particularly preferred embodiments, the minimum distance is between 0 mm and 5 mm, such as 0 mm, 1 mm, 2 mm, 3 mm, or 5 mm.

[0150] In one aspect, step b) of the method according to any one of the embodiments set forth herein and in the claims includes fixing the plurality of fragments or accessions provided in step a) to at least one support element such that the minimum distance between each of the fragments or accessions and at least one other of the fragments or accessions, i.e., at least an adjacent fragment or accession, is 0 cm to 2 cm, 0 cm to 1 cm, preferably 0 mm to 5 mm, and more preferably 0 mm, 1 mm, 2 mm, or 3 mm.

[0151] In certain embodiments, each substrate to which at least one affixed individual according to any one of the variations described herein is attached, affixed, or immobilized is fixed to a support element such that the minimum distance between the affixed individual on that substrate and at least one affixed individual attached or affixed to another of the substrates is 5 mm or less.

[0152] In a specific aspect, step b) of the method according to any one of the embodiments set forth herein and in the claims then comprises fixing the substrates provided in step a) to at least one support element, each of which has fixed thereto at least one of the precious coral recruits according to any one of the variations described herein, such that the minimum distance between each recruit attached, immobilized, or affixed to each of the substrates and at least one recruit attached, immobilized, or affixed to a different substrate is 0 mm to 5 mm.

[0153] According to a preferred embodiment of the present invention, each fragment or accession fixed to a support element is in contact with at least one adjacent fragment or accession fixed to a support element at least at one point. In other words, according to a preferred embodiment, each fragment or accession fixed to a support element is positioned so that the minimum distance between it and at least its adjacent fragment or accession is 0 mm.

[0154] According to one embodiment of the invention, in step b), the fragments or adducts, in particular the substrates to which the adducts are respectively attached, attached or immobilized, are fixed to a plurality of separate support elements, with the proviso that at least two fragments, adducts or substrates are fixed to each support element.

[0155] One support element suitable for use in the method according to any one of the embodiments of the present invention is any material known in the art suitable for securing or "attaching" corals or their recruits.

[0156] Suitable supports for use in step b) of the method according to any one of the embodiments described herein include materials selected from cement, ceramic, marble, travertine, plastic, in particular polyvinyl chloride (PVC), or other materials known in the art capable of fixing precious coral fragments, or combinations of these materials.

[0157] According to a preferred embodiment, the support element or elements used in step b) of the method of the present invention are obtained by mixing type 2 Portland cement with sand and fresh water, preferably in a ratio of about 1:4:1. The resulting mixture is cast to obtain the shape of the desired support, and the resulting shape is air-dried for at least 24 hours.

[0158] According to a further aspect of the invention, the one or more support elements used in step b) of the method of the invention comprise or consist of one or more seabed elements or surfaces suitable for the anchoring or settlement of corals, said seabed elements or surfaces being selected in particular from coral rock, carbonate rock, detrital skeletons of coral colonies, or combinations thereof. Seabed elements or surfaces suitable for use as support elements according to the invention have non-biological conditions suitable for the growth of precious corals.

[0159] The shape and size of the support element(s) used in step b) of the method of the present invention can be varied according to the particular needs, in particular the number of fragments or recruits selected and the type of nursery used for the culture process. Those skilled in the art will be able to select the optimal shape and / or size for the support element(s) depending on the number of fragments or recruits used, the desired final size of the resulting precious corals, colonies or recruits, and the type of nursery used.

[0160] According to one aspect of the present invention, the support element(s) used in step b) of the method according to any one of the embodiments described herein are generally disc-shaped. By way of example only, the method according to the present invention may provide for the use of one or more disc-shaped support elements with a diameter of 10 cm, known in the art as "cookies."

[0161] Fragments of precious coral recruits, particularly substrates to which recruits according to any one of the variations described herein are attached, affixed, or immobilized, can be secured to one or more selected support elements by using suitable securing means according to any one of the variations known to those skilled in the art. Non-limiting examples of securing means suitable for use in the methods of the present invention include glue, or synthetic or natural adhesives, such as cyanoacrylate glue or epoxy resin, as well as other adhesives or stickies that are resistant to marine environments, threads, bands, suitable supports made of PVC or other materials, or other mechanical securing means. Preferably, two-part epoxy resin or cyanoacrylate glue, or other adhesives that are resistant to marine environments, are used to secure the fragments or substrates to the support element.

[0162] Alternatively, the substrate to which the fragment, or the affixed individual according to any one of the variations described herein, is attached, affixed or immobilized may itself be fixed to one or more support elements as the fixing means.

[0163] In particular, according to one embodiment of the method of the present invention, the at least one support element used in step b) is made of fishing line or nylon line, or the same material as that used to secure the fragments at the desired mutual distance. In this case, the selected fragments can be connected to each other using the same support element, i.e., fishing line, as a fastening means, provided that the minimum mutual distance between each fragment and at least one other fragment when secured falls within one of the ranges specified in the present specification and claims. Furthermore, as described in detail below, the fragments connected by fishing line can be suspended in any device known in the art for cultivating or propagating corals in the sea, tanks, or aquaria (e.g., dendritic propagation beds), or can be inserted between the meshes of one or more ropes commonly used in the art. The ropes generally have a braided structure. Thus, the fragments are inserted between the braided mesh and are held in place by the tension of the rope itself.

[0164] In one aspect of the present invention, in step b) of the method according to any one of the embodiments described herein, each fragment is fixed to the at least one support element via its end. The end is preferably the end corresponding to the oldest part of the fragment, for example, the end resulting from cutting the fragment from the tip or apical end of the precious coral colony. In this configuration, a first end of each fixed fragment is fixed to the at least one support element, and a second end is a free end. The first end includes a base for being supported or fixed on a support element.

[0165] According to one aspect of the present invention, in step b) of the method according to any one of the embodiments described herein, the selected fragments are fixed to the at least one support element in a vertical position, in particular so that the longitudinal direction of the recruitment or fragment is perpendicular to the surface of the support element. According to a preferred aspect, in step b) of the method according to any one of the embodiments described herein, the ends of each fragment resulting from cutting or trimming the fragments obtained from the existing colony of precious coral are fixed to the at least one support element so that each fragment is in a vertical position.

[0166] In a further aspect, when the method of the present invention provides for the use of precious coral fragments as disc-shaped colonies (preferably, for example, colonies having a diameter of 5 mm to 15 mm, a thickness of 2 mm to 5 mm, and an age of 2 to 4 years), the colonies are fixed to the at least one support element in a vertical position. In particular, the colonies are fixed so that a portion of the periphery of each colony is in contact with the support element. In such a configuration, since the contact area between the colonies and the support element is very small, it is preferable to use an adhesive that can ensure suitable fixing resistance to the support element, such as a two-component resin, PVC, or a suitable support made of another suitable material, to fix the colonies.

[0167] In one embodiment, the fragments or individuals according to any one of the embodiments described herein are fixed to the at least one support element in a generally parallel position, in particular such that the longitudinal extension directions of each fragment are parallel to one another.

[0168] In a further embodiment, selected pieces can be secured to a support element so that they are stacked on top of each other, for example, by using mechanical fasteners such as PVC rods secured to the support element in a vertical position.

[0169] According to one aspect of the present invention, in step b) of the method according to any one of the embodiments described herein, the substrate to which the fragments or selected individuals, in particular the individuals according to any one of the variations described herein, are attached, affixed or immobilized, is fixed to said at least one support element so that the fragments or selected individuals form a motif or pattern.

[0170] In other words, precious coral fragments or accessions, or substrates to which said accessions according to any one of the variations described herein have been attached, affixed, or immobilized, can be fixed to one or more selected support elements to form a particular arrangement. Such motifs and arrangements can be selected and predetermined by those skilled in the art depending on the type of individual, accession, fragment, or substrate selected, and on the desired final shape of the precious coral, colony, accession, or chimera.

[0171] Preferably, the fixed pattern or arrangement is linear, striped, beam-like, triangular, rectangular, or circular, particularly any shape suitable for plates such as cameos and gem coral pellets, as well as other shapes of interest in the jewelry field.

[0172] By way of example only, the addenda, fragments, or substrate to which the addenda are attached or affixed may be secured to respective support elements in sets of two to form a line, in sets of three to form a triangle, or in sets of four to form a square.

[0173] In a preferred embodiment of the present invention, the selected fragments are secured to form one or more lines or one or more beams. This arrangement is useful for forming cameos and / or plate-shaped gem coral pellets, which are intended for jewelry, and the final size can be varied depending on the number of fragments used. By way of example only, securing seven pieces of gem coral in a line can produce a cameo with a long axis of approximately 4 cm and a short axis of approximately 2.5 cm, while securing seven pieces in a beam (six pieces arranged in a hexagon and one piece in the center) can produce pellets 7 mm to 8 mm wide (with a 1 mm interfragment spacing). Furthermore, securing gem coral fragments with a colony diameter of approximately 15 mm and a height of approximately 5 mm can produce small colonies of red coral, resulting in pellets 10 mm to 12 mm wide.

[0174] The fragments or recruits can be secured to the support element to form shapes and patterns not found in naturally occurring corals, such as wavy plates, letters, numbers, and other fantastical shapes.

[0175] As mentioned above, each pattern or specific arrangement of fragments or recruits fixed to the support element can be achieved by using fragments or recruits from the same colony (homologous fusion), or fragments or recruits from the same species but from a different colony (heterologous fusion), or fragments or recruits belonging to different species (interspecific fusion), or recruits less than 6 months old from the same species but from a different colony (chimerism).

[0176] In step c) of the method according to any one of the embodiments described herein, the substrate to which the fragments or accessions, particularly the accessions according to any one of the variations described herein, are attached, affixed, or immobilized is suitably fixed to one or more support elements according to a specific arrangement devised by the inventors, and transplanted into seawater, where it is cultured for a period of time sufficient to promote fusion between adjacent fragments or accessions, resulting in the formation of a precious coral, colony, accession, or chimera having a desired shape and size, particularly a desired diameter.

[0177] Optionally, in order to immobilize or hook the support element(s) used in step b) to a suitable device for culturing / growing the fragments in the sea and / or in a controlled environment, such support element(s) may be provided with appropriately drilled holes or mechanical hooking means, as exemplified below.

[0178] Alternatively, any suitable fixation support or fixation method known in the art can be used, for example, the fragments or accessions can be fitted directly into small PVC tubes which are then fitted into the mesh of plastic netting, or the fragments or accessions can be glued to garden stakes which are then fitted into plastic netting.

[0179] The use of mechanical hooks is particularly advantageous when there is excessive contamination or a large amount of microalgae, for example in fish farms during the summer. Indeed, supports fixed to the nursery bed by mechanical means can usually be easily removed from the nursery bed, shaken to decontaminate the colonies, and then fixed back to the nursery bed. Alternatively, the water can be stirred by hand, or a lance can be used to generate a water current using compressed air from a cylinder.

[0180] Preferably, a numbered tag, e.g., a metal tag, can be attached to each support element used in step b) to allow identification of the samples being cultured and to record the selected fixation arrangement, type of fusion and / or start date of culture, as well as other information that may be relevant to the coral species used in the method of the present invention.

[0181] According to one aspect of the invention, prior to carrying out step c) of the method according to any one of the embodiments described herein, the fragments or accessions suitably fixed in step b) are transferred in seawater, preferably to a "quarantine" or "accumulation" tank, where they are maintained under controlled conditions, preferably at a temperature ranging from 2° C. to 30° C., preferably from 16° C. to 20° C., for a period of 7 to 30 days, preferably at least 30 days. This period is long enough to significantly reduce the chance of a subsequent outbreak of disease in the nursery.

[0182] Such procedures are useful for better assessing the condition and vitality of selected fragments or recruits and for accumulating an appropriate number of fragments or recruits before placement in a suitable culture environment, such as open water. Ultimately, they are useful for reducing the number of dives required to properly place fragments or recruits in the selected culture area. If necessary, targeted therapy or the use of disinfectants of the type used in aquaria, according to the manufacturer's formulation and concentration, can also be used.

[0183] The seawater used in step c) for culturing fragments or recruits of precious coral colonies according to any one of the embodiments described herein can be artificial seawater or natural seawater, preferably natural seawater.

[0184] According to one aspect of the invention, step c) of the method according to any one of the embodiments described herein comprises transplanting the plurality of fragments or accessions fixed to the at least one support element into a host environment selected from: Open waters, and / or Controlled marine environments selected from natural or artificial bodies of water, e.g., fish farms or protected marine areas; tanks, e.g., land-based tanks; or aquaria.

[0185] According to a preferred aspect of the present invention, step c) of the method according to any one of the embodiments described herein comprises at least the following steps: c') The fragments or accessions fixed in step b) are cultured in seawater at a temperature ranging from 2°C to 30°C, preferably from 16°C to 22°C, in a controlled marine environment selected from natural or artificial bodies of water, tanks or aquaria, until fusion of the fragments or accessions is obtained. c'') The above fused fragments or fused individuals are transplanted into open waters.

[0186] According to one aspect of the invention, when transplanting fragments or recruits into open waters, the at least one support element and the fragments or recruits attached thereto are placed at a depth below the summer thermocline of the selected water area, optionally using a suitable culture device as exemplified below.

[0187] Preferably, in open waters, the fragments or recruits are placed at a depth of 15 to 100 meters. This is important to ensure that the precious coral recruit fragments are maintained at a fairly constant temperature between 2 and 30°C throughout the entire cultivation and propagation process. Depending on the origin of the selected precious coral species, those skilled in the art can identify the optimal temperature range in which the fragments or recruits should be maintained during cultivation step c) and, in the case of underwater cultivation, determine the ocean depth at which the immobilized fragments or recruits should be maintained at the desired temperature. By way of example only, red coral fragments are preferably maintained at a temperature between 16 and 22°C, while fragments of Hemicorallium laauense, which often inhabit deep waters, are preferably maintained at a temperature between 5 and 12°C.

[0188] In order to ensure that the coral recruit fragments subjected to cultivation step c) in the sea are maintained at a suitable temperature within the ranges specified in the present specification and claims, the location, i.e. the depth at which said at least one support element and then the fragments or recruits secured thereto are placed, may be varied throughout the year based on seasonal thermocline fluctuations in the selected sea area.

[0189] By way of example only, in the case of red coral, the fragments or recruits secured to the at least one support element may be placed in a selected area at a depth of 18 metres in winter and then at a depth of 40 metres in summer.

[0190] 40 metres is the maximum depth that amateur divers can safely dive to using air, and the depth that professional divers can safely reach without using gas mixtures or decompression stages.

[0191] Alternatively, particularly in the case of precious coral species that live only in the deep sea (aphotic zone), it is possible to place the nursery in the depth zone where the species to be used live and then recover them at the end of the cultivation. In this case, the corals are preferably placed in the nursery before diving according to the above-mentioned configuration, and the nursery is preferably provided with a device, such as a metal ring or other device known in the art, that can be used to hook the corals for recovery.

[0192] Depth buoys used to ensure good seedbed positioning must be designed to prevent implosion at the selected water depth. There are several materials, capacities, and pressure resistance capabilities available on the market, and a skilled artisan can select a suitable product based on the manufacturer's technical specifications and the depth at which the seedbed will be placed.

[0193] Since corals feed on suspensions carried by ocean currents (suspension feeders), this parameter is an important factor for the culture of such species.

[0194] In the case of transplantation / cultivation in open waters, it is preferred to place the fragments or accessions fixed to said at least one support element in a water area with a strong enough water current suitable to ensure a sufficient supply of nutrients, which can be assessed by a person skilled in the art based on the species and the chosen water area.

[0195] When transplanting / cultivating fragments or recruits into a controlled marine environment, such as an artificial or natural body of water, a tank or aquarium, it is preferable to generate a current of seawater in such an environment that circulates around one or more support elements to which the fragments or recruits are fixed.

[0196] According to one aspect of the invention, food materials for growth can be added to the stream, examples of which are small planktonic organisms, non-living particulates, dissolved organic matter, or combinations thereof.

[0197] The seawater used in step c) for culturing precious coral fragments or recruits according to any one of the embodiments described herein preferably has a certain salinity, in particular a concentration of 28‰ to 40‰, which concentration varies in relation to the precious coral species of interest. Those skilled in the art will be able to identify the most suitable salinity range based on the various precious coral species selected.

[0198] As mentioned above, in one aspect of the invention, step c) of the method according to any one of the embodiments described herein comprises transplanting the plurality of fragments or recruits fixed to the at least one support element into a controlled marine environment representative of a fish farm. Placement in a fish farm is particularly advantageous as it can take advantage of concentrated nutrients produced by organic residues from the farm to increase the rate of biomineralization of the colony.

[0199] According to one aspect of the present invention, in step c) of the method according to any one of the embodiments described herein, the at least one support element is fixed to or placed in an apparatus suitable for culturing corals in seawater and / or saltwater.

[0200] According to a further embodiment, the one or more support elements used in step c) of the method of the present invention may be fixed in one or more containers (known in the art as "trays") suitable for being fixed to or placed in the selected culture device.

[0201] For fastening the support elements to the tray or directly to the selected cultivation device, as well as for the possibility of fastening the same tray to the selected device, conventional fastening means known in the art, such as fishing line, plastic bands or mechanical fastening means, can be used, with care taken to remove excess parts of such fastening means in the case of submerged cultivation in order to avoid algae accumulation.

[0202] Apparatus usable in step c) of the method according to any one of the embodiments described herein for culturing fragments or recruits immobilized in seawater and / or seawater may include any apparatus suitable for culturing or growing corals.

[0203] Devices particularly suitable for culturing corals in sea and / or saltwater include "coral nursery" devices, which are coral nurseries known in the art. Non-limiting examples of such devices include various types of nurseries, such as floating nurseries, fixed nurseries, electric or electrified nurseries, string nurseries, and / or predation-resistant nurseries.

[0204] Depending on the host environment selected for the cultivation of the fragments or recruits, and the particular needs, funding, and equipment available, one skilled in the art will be able to identify the most suitable type of equipment or nursery to use in the cultivation in seawater of the particular type of coral fragments used in the methods of the present invention.

[0205] According to a preferred aspect of the present invention, in step c) of the method according to any one of the embodiments described herein, the at least one support element is fixed to a floating mini-nursery, and the cultivation of coral fragments or coral recruits is carried out in open water. This floating mini-nursery is a very inexpensive and practical device. It can be assembled on land, requires little maintenance, and significantly reduces the problem of algae and debris accumulation. This type of nursery can be easily raised to a more comfortable working depth and then lowered back to the selected cultivation depth. This greatly simplifies underwater operations and is particularly suitable for installation at dangerously high depths where only short dives are permitted.

[0206] Preferably, the floating mini-bed consists of a substantially rectangular pallet, in particular made of plastic and measuring 120 cm x 100 cm, anchored to the seabed, with buoyancy buoys fixed to the corners of the pallet, suitable for floating at a predetermined distance from the seabed. In use, the top surface of the pallet of the fusion mini-bed faces the water surface and the bottom surface faces the seabed.

[0207] The one or more support elements are preferably secured to the upper and / or lower surfaces of the nursery pallet. Securing the one or more support elements to the underside of the pallet allows the fragments secured to the lower surface to be positioned "upside down," mimicking the preferred growth conditions of gem corals, such as red coral, which grow on sediment-free cave ceilings and under rock corners.

[0208] According to one aspect of the present invention, the method according to any one of the embodiments described herein further comprises at least a step of examining the fragments or participants immobilized on the at least one support element cultured in step c).

[0209] Testing is preferably carried out at regular intervals, for example weekly or monthly, which is useful not only to monitor the progress of the culture, but also to facilitate cleaning of fragments, inoculations, support elements, and equipment, if used.

[0210] In fact, visual inspection allows us to assess the condition of the corals, the progress of fusion, and the structural integrity of the equipment, if used, allowing us to remove as many corals as possible that show signs of necrosis, tissue lesions, or pathology, and to prevent their spread.

[0211] Inspection is also useful for cleaning algae and sediment, removing fouling and potentially predatory sponges or their larvae, and performing temporary maintenance if the culture equipment used has structural defects, as well as for performing structural modifications to said equipment. As corals grow and algae and debris accumulate, the flotation of the nursery beds may change, and over the course of a long period of culture, this flotation may need to be corrected.

[0212] For example, a soft bristle brush or toothbrush can be used to clean algae and debris, while a knife or other tool known in the art can be used to remove sponges and encrustations.

[0213] To facilitate inspection and / or cleaning procedures when the cultivation step c) is carried out at a suitable depth, e.g. 40 m at sea, it is generally desirable to transfer the fragments or recruits fixed on the at least one support element or any cultivation device that may be used to a shallower working depth, e.g. 9 m.

[0214] The duration of step c) of the method according to any one of the embodiments described herein or in the claims is preferably 4 years or less, more preferably 2 years or less, even more preferably 1 to 2 years, even more preferably 4 to 8 months.

[0215] A further aspect of the present invention is represented by a precious coral, or a precious coral colony, recruit, or chimera, obtainable by a method according to any one of the embodiments described herein.

[0216] The following examples are provided for the purpose of more clearly illustrating the methods exemplified herein, and such examples should not be construed as limiting the scope of the foregoing description and the claims that follow. [Example]

[0217] Example 1: Cultivation of red coral fragments by fusion We have devised a method for culturing red coral fragments in the sea, which involves the following materials and steps: 1) Preparation of coral fragments

[0218] As starting material, fragments of red coral measuring 2.5 cm to 5 cm were used.

[0219] The fragments were searched for by licensed, professional fishermen, who typically "remove" the "tips" of the corals they catch - the tops of the twigs that are too thin to be processed into gemstones. This is because, although the commercial value of the tips is very low, they count towards daily and annual fishing quotas. Tips are currently a waste product that is recommended, but not mandatory, to be discarded from boats in the field by the Mediterranean General Fisheries Commission.

[0220] These coral tips are "ideal fragments" that allow us to avoid directly collecting fragments found in nature.

[0221] Selected fragments were stored in seawater at temperatures between 16 and 22°C and separated based on their locality to avoid confusion. To keep them separate, perforated plastic containers (i.e., molds), commonly used in cheese processing, were used. The molds were immersed in larger containers filled with chilled seawater using improvised ice packs, taking care to avoid direct sunlight.

[0222] Bonsai shears were used to cut the pieces.

[0223] All materials used were first disinfected with 70% ethanol and rinsed thoroughly with fresh water. Sterile surgical gloves were used when handling the fragments. 2) Fixing the fragments to the support element

[0224] The selected red coral fragments were fixed onto 10 cm diameter discs (cookies) with four holes for later fixing into the fusion nursery.

[0225] The cookies were made from Type 2 Portland cement, river sand, and fresh water (approximately 1:4:1) and allowed to dry for at least 24 hours.

[0226] Numbered metal tags were attached to the supports to record the fixation location, type of fusion, and date of culture initiation.

[0227] To achieve fixation to the support, each "cut" piece was properly dried using a clean cotton cloth and glued to the support with cyanoacrylate adhesive.

[0228] Again, the pieces were handled with sterile gloves during this step.

[0229] The sections were carefully inspected visually to assess the presence of lesions, tissue necrosis, and polyp vitality, if possible. If defective, they were discarded.

[0230] The pieces were fixed to each support in sets of 2 (lines), 3 (triangles), or 4 (squares) at 0, 1, or 2 mm intervals. Each configuration was replicated three times.

[0231] To achieve larger cameos and pellets, we fixed a set of seven fragments arranged in a line (0mm and 1mm intervals) and a set of seven fragments arranged in a beam (0mm and 1mm intervals). Each configuration was repeated three times.

[0232] Each construction was performed using both fragments from the same colony (homologous fusions) and fragments from different colonies (homologous fusions).

[0233] Finally, 30 single pieces were used as references (10 pieces for each of the seedbeds used), with three repeats for each configuration for three different seedbeds.

[0234] Once the fragments are fixed onto the cookies, they are stored in refrigerated seawater at 16°C to 22°C until they are placed in fusion nurseries.

[0235] To simplify the undersea procedure, each cookie was fixed on board the ship to a 4x4 (30cm x 50cm) plastic electrowelded net tray, which was then fixed to the fused nursery bed. Alternatively, plastic net trays with different mesh sizes (not too fine to prevent excessive algae accumulation) and made from different materials could be used, or the cookies could be fixed directly to the nursery bed.

[0236] For fixation, we used 2.5 mm plastic bands, taking care to remove excess to prevent algae buildup, but alternatively, bands of different sizes, fishing line, or mechanical fixation methods can be used. 3) Subsea culture of fragments using floating fusion nurseries

[0237] The fusion of selected fragments was carried out in three miniature nurseries floating for underwater cultivation. These are very cheap (materials cost 100 euros), practical and can be assembled on land, require little maintenance and significantly reduce problems with predators, diseases, algae and debris buildup. They can then be easily raised to a more comfortable working depth and then returned to the cultivation depth. This significantly reduces underwater work and makes them particularly suitable for installation at high depths where diving time is short and dangerous.

[0238] Each nursery bed consisted of a plastic pallet (120 cm × 100 cm) with four 1-liter buoyancy buoys fixed to its corners and secured to the bottom with 50 kg cement weights.

[0239] Cookies are typically placed on top of nurseries, but red corals tend to grow upside down on the ceilings of caves or under rock corners where sediment is absent. Therefore, one of the three nurseries used in the study was a box pallet assembled inverted to simulate a cave. Coral fragments were hung upside down, considering that this would likely result in better results in direct transplantation tests. This configuration not only replicates the "preferred" position of red corals, but also avoids the generation of sediment. The edges of the box were perforated to allow water flow while protecting the corals.

[0240] Alternatively, different sizes, different materials, different means of anchoring to the seabed, and any type of nursery known or suitable for coral propagation (floating, fixed, electric, predation-resistant, etc.) can be used depending on the site and project contingencies.

[0241] In the sea area chosen for the experiment (the Mediterranean Sea), surface temperatures can exceed 26°C in summer.

[0242] However, the red coral used in the experiment cannot tolerate temperatures above 22°C for long periods of time.

[0243] Therefore, the nursery was placed below the region's summer thermocline (a zone of several metres with a steep temperature gradient of up to 10°C), where temperatures are more constant and do not exceed 20°C even in summer.

[0244] In the past, this area was known for its shallow thermocline (around 12 m) in summer, and in fact, corals were already thriving at depths of 18 or 20 m.

[0245] However, in recent years, both fishermen and biologists have reported a deeper thermocline (35 m) or its absence, causing mass mortality (MME) of gorgonia (a coral relative) and red corals above 35 m.

[0246] One nursery was kept at 40 m, while the other was positioned along the seasonal thermocline. The simulated cave was fixed at 18 m and lowered to 40 m in summer.

[0247] In fact, corals in this area also grow above 35m, but only in dimly lit caves and canyons.

[0248] Furthermore, 40m is the maximum depth at which non-professional divers can safely perform air diving, and is a fairly safe depth for professional divers, reaching depths without the use of mixed gases or decompression.

[0249] Furthermore, corals feed on suspended nutrients carried by ocean currents (suspension feeders), meaning that water currents are more important to them than the concentration of nutrients (phytoplankton and microplankton).

[0250] To ensure suitable biological conditions, nursery placement depths were chosen at locations where the presence of colonies had been recorded by coral experts. 4) Preliminary results

[0251] Fixing the coral fragments at a selected distance promoted fusion between adjacent fragments, and in initial qualitative experiments, we observed a higher survival rate for fused fragments than for single fragments used as references.

[0252] Without wishing to be bound by theory, this phenomenon can be explained by considering that coral growth and survival rates depend more on size and polyp number than on age, so a higher number of polyps and the larger size of fused fragments compared to single fragments may explain this effect.

[0253] As with other modular organisms, size appears to be more important than age for many biological processes in corals, and size rather than age can sometimes better explain coral developmental patterns.

[0254] At the same age, larger corals have higher growth and survival rates, are more likely to reproduce (corals reach sexual maturity at a certain size, not at a certain age), are better able to adapt and recover, are more efficient at photosynthesis, and have more available energy because they are exposed to more water and can capture more prey. Example 2: Cultivation of red coral colonies by fusion

[0255] We devised a method for culturing fragments of red coral consisting of 2- to 4-year-old recruits with any genome (any parent) by homotypic and heterotypic fusion. This method involves the following materials and procedures. 1) Larval recruitment and incubation on seedbeds

[0256] For example, new colonies of red coral were generated from 2- to 4-year-old red coral recruits with largely random genomes by fusion according to the methods involved in this application.

[0257] The use of recruits of this kind has great advantages from an ecological point of view, and in our opinion, the recruitment of wild larvae to attachment substrates is the most sustainable way to collect and supply the fragments used in the method of the present invention.

[0258] Alternatively, recruits may be collected directly from the wild marine environment to provide a natural substrate, or may be produced according to any method generally known to those skilled in the art.

[0259] As mentioned above, the preferred method is to collect adults from wild colonies, breed them in suitable aquaria, and immerse them in the aquaria with a support on which the young can settle. All the young thus produced will be of the same parent and will have compatible genes that allow homozygous fusion, which, as is well known, is more stable than homozygous fusion. However, this method requires a suitable system.

[0260] In the case described herein, for example, larvae were captured in a wild marine environment approximating a surface population of red coral at depths of 35 to 55 meters, and the larvae have essentially any genetic makeup that sporadically forms isomorphic pairs. In the case described herein, colonies formed by isomorphic fusion are only sporadically formed.

[0261] By way of example only, juvenile recruitment was carried out on 30cm x 30cm marble tiles with a 1cm thick hole in the centre, which were secured to the vicinity of red coral breeding areas at depths of 35mt to 55mt using two-component resin via 1cm galvanised threaded rods.

[0262] Alternatively, any method known in the art can be used for juvenile recruitment. Tiles were placed in May-June and harvested in October-November. Recruits attached to the capture substrate ranged in age from approximately 0 to 6 months.

[0263] The recruitment substrates with attached recruits were assembled on floating mini-nursery beds of the type described above, located near aquaculture facilities with an operating depth of 25 mt, on the seabed at depths of 35 to 40 m.

[0264] Once the explants reach a thickness sufficient for detachment, they are removed from the explant substrate and are ready for synculture. In the examples described herein, seed bed cultures continued for 2 to 4 years.

[0265] The seedbed production cycle described as an example herein lasts from two to four years, but this only affects the start of the seedbed, since juveniles are harvested annually, and year-round harvesting of juveniles will result in annual production after four years. Production depends primarily on the total area dedicated to juvenile establishment. 2) Cultivation by fusion of red coral fragments produced in the seed bed according to the method of the present invention 2.1) Selection of recruits and fixation to one support element

[0266] Disc-shaped red coral colonies were used as starting fragments for the fusion culture method. The colonies were 2 to 4 years old, approximately 5 to 15 mm in diameter, and approximately 2 to 5 mm in height or thickness (measured at the center of the base). Alternatively, fragments of younger precious corals could be used, provided they were large enough to be detached from the growth substrate used. Older fragments could also be used, but would require a longer incubation period in the seed bed.

[0267] By way of example only, colonies were fixed to rectangular travertine supports measuring 10 cm x 10 cm and 0.5 cm thick. The shape and size of the support elements were selected to optimize spacing in the nursery. The selected colonies were fixed in a vertical position on each support element, in four rows with parallel bases, spaced apart by less than 1 mm, and more preferably 0 mm. In particular, the edges of each disk-shaped piece were fixed to the support element using a two-component resin. A single recruit was also fixed to each support for reference.

[0268] Alternatively, other configurations can be used, provided that the distance between adjacent pieces is less than 5 mm, preferably less than 1 mm, and even more preferably 0 mm. For example, the disk-shaped pieces can be stacked on top of each other (like a voltaic cell) using a mechanical fixture consisting of four polyvinyl chloride rods fixed perpendicularly to a support to form a rectangle whose diagonal is slightly smaller than the diameter of the colony. 2.2) Culture by allogeneic hybridization in floating seedbeds

[0269] The colonies thus fixed are then arranged in the same nursery beds previously used for the seedbeds, according to the aforementioned configuration, and cultured in seawater. The resulting colonies are cylindrical with irregular sides. For tightly fixed colonies, the expected culture period is 18 to 24 months, which is sufficient time for the minimum diameter of the cylinders to reach the desired size. The final product is represented by small colonies of red coral, with pellets ranging in size from 8 mm to 15 mm. 15 mm represents the maximum size obtainable from pellets produced by this particular embodiment. Example 3: Production of large chimeras or recruits for use in producing precious corals according to the method of the present invention

[0270] We have devised a method for culturing large-sized chimeras or recruits of precious corals produced by chimerism or allomorphic fusion from recruits of red corals that are less than 6 months old, more preferably less than 3 months old, and even more preferably less than 1 month old.

[0271] Replacing the substrate monthly, quarterly, or biannually can ensure that captured recruits are young enough to chimerize.

[0272] The size of such recruits is close to zero, and they are merely point-like attachments. 1) Juvenile recruitment

[0273] Precious coral juveniles under six months old are too thin to be detached from the attachment substrate. To achieve juvenile recruitment, it is necessary to use a substrate that can be cut with millimeter or submillimeter accuracy, and to use recruits attached to a recruitment substrate in the method of the present invention.

[0274] For example, a 1mm thick PVC sheet measuring 30cm x 30cm can be used. Alternatively, substrates of different sizes, thicknesses, and materials can be used, provided that the material is conducive to coral attachment and settlement and can be cut to millimeter or sub-millimeter accuracy. Examples include glass, steel, and marble.

[0275] The PVC sheet is attached to a rigid support element with a centrally perforated hole using a marine-resistant adhesive, preferably applied only to the edges of the sheet and around the hole, allowing it to be easily removed from the rigid support element later using a cutter or other suitable tool.

[0276] The substrate is preferably a 30x30 marble tile with a hole in the center, and the joining substrate is secured to the marble substrate with a marine environment-resistant adhesive applied to the edges and around the hole in the substrate.

[0277] A rigid support is not essential, but is practical and an alternative is PVC, but only greater than 1mm thick, or any other type of support that can be coral-received and cut to millimetre accuracy.

[0278] Juvenile recruitment was conducted in a natural marine environment near the surface area of ​​red coral at depths of approximately 35 to 55 meters. Therefore, the collected juveniles have virtually any genome. Alternatively, for larval recruitment, tank spawning is preferred because it allows for the generation of larvae derived from a single parent. However, any technique known in the art for promoting larval settlement is effective. 2) Generation of recruits or chimeras

[0279] Once the rigid support on which the attachment substrate with attached or attached larvae is secured is recovered, the PVC substrate is removed from the rigid support and cut into small pieces, each containing at least one recruit, particularly at least one recruit located at a suitable distance from one of the edges of the substrate.

[0280] A suitable distance from the edge means that when a substrate containing one accession is fixed to a support for cultivation in a nursery, the distance between each accession and at least one other accession is less than 10 mm, more preferably less than 3 mm, even more preferably less than 1 mm, or 0 mm.

[0281] By way of example only, in generating chimeras, PVC sheets were cut into 2 cm x 2 cm square pieces, with one recruit less than 1 mm from one edge and approximately 1 cm from the adjacent edge. To generate larger recruits for use in the method of Example 2, PVC sheets were cut into 2 cm x 2 cm square pieces, with one recruit approximately 1.2 mm and 3 mm from one edge and approximately 1 cm from each adjacent edge.

[0282] The PVC pieces obtained as described above were fixed to a suitable support as pairs of two or four pieces per square by tightly contacting the edges of the substrate pieces near the joining individuals present on the substrate pieces, taking care not to leave any steps or gaps between the PVC pieces.

[0283] The shape, size and material of the support for fixing each PVC piece is not relevant in this example. As an example, a 10 cm x 10 cm travertine tile was used with four holes drilled for fixing it to the fused bed with plastic bands. Alternatively, variations of the above or those already known in the art can be used.

[0284] The supports were then cultured in a fusion bed.

[0285] As fusion beds, floating mini-beds were used as already described and arranged in the fish farming facility in the manner already described, or any variant known in the art or already described above could be used.

[0286] Since the genotype of recruits in the present invention is almost arbitrary, the majority of recruits in the present invention are allogeneic pairs (different parent colonies), with sporadically observed allogeneic pairs (same parent colonies).

[0287] Allogamy produces new recruits of red corals with larger polyp size and number, whereas allogamy produces chimeras of red corals.

[0288] The required incubation period in the fusion nursery before accessions can be used for cultivation using the method of the present invention is 4 years, preferably 2 to 3 years.

[0289] Alternatively, given that homotypic fusion and chimerism result in stable colonies, accessions generated according to the fusion methods described herein can be kept in culture for much longer periods, even for 10 years or more, and then used for transplantation or as starting fragments for the methods of the invention or for commercial purposes.

[0290] The fragments and chimeras produced in this way, consisting of two or four accessions fused together, are larger and have greater biomass and polyp numbers than the fragments produced in the seed bed described in Example 2 using fragments composed of a single accession, allowing the maximum diameter of the seed bed described above to exceed 1.5 cm, generally improving performance.

[0291] Based on what is known about chimeras, this particular example could be used to promote horizontal evolution between precious corals and common corals through chimerism, and perhaps even induce horizontal evolution in their variants with known genotypes. Indeed, it has been hypothesized that it may be possible to further enhance resistance by selecting corals with specific resistance to climate change (e.g., corals that have survived mass mortality or bleaching events) and chimerizing these corals with each other. For the first time, the method presented here makes it possible to promote chimerism among precious coral recruits using any genome, or genomes selected to be highly advantageous for coral repopulation projects.

[0292] Furthermore, given that coral hybrids or interspecies hybrids exist in nature, it is possible to hypothesize that this particular embodiment of the present invention may be used to promote interspecies chimerism, where chimerism promotes interspecies fusion between young recruits, and subsequently produce interspecies hybrids of corals, particularly gem coral hybrids.

Claims

1. 1. A method for producing precious corals, or precious coral colonies, recruits, or chimeras, comprising: a) providing at least two fragments or recruits of existing precious corals belonging to the Corallinaceae family; b) fixing said fragments or said members to at least one support element so that the minimum distance between each of said fragments or members and at least one other of said fragments or members is between 0 cm and 2 cm, preferably between 0 mm and 5 mm; c) culturing the fragments or the accessions fixed in step b) in seawater at a temperature ranging from 2°C to 30°C, preferably from 16°C to 22°C; A method comprising:

2. 2. The method of claim 1, wherein the at least two fragments or the at least two recruits belong to one or more species of precious coral selected from red coral (Corallium rubrum), red coral (Corallium japonicum), pink coral (Pleurocorallium elatius), white coral (Pleurocorallium konjoi), Pleurocorallium secundum, Hemicorallium regale, Hemicorallium laauense, and water coral (Hemicorallium sulcatum).

3. The method according to claim 1 or 2, wherein the at least two fragments or the at least two recruits belong to the same or different species of the precious coral belonging to the Corallinaceae family.

4. 4. The method of any one of claims 1 to 3, wherein the at least two fragments or the at least two accessions are red coral fragments or accessions.

5. 5. The method according to claim 1, wherein the at least two fragments are apical portions of precious coral obtained from branches of the precious coral colony.

6. 10. The method of claim 9, wherein each of said segments has a length of from 0 cm to 10 cm, more preferably from 2.5 cm to 5 cm.

7. 5. The method according to claim 1, wherein the at least two fragments are colonies of precious coral belonging to the Corallinaceae family and are aged between 0 and 10 years, more preferably between 6 months and 4 years, and even more preferably between 2 and 4 years.

8. 10. The method of claim 1, wherein the at least two fragments are colonies of precious coral belonging to the Coralline family and are approximately disc-shaped.

9. 10. The method of claim 9, wherein the colonies have an average thickness of 0 mm to 20 mm, more preferably 2 mm to 5 mm, and / or an average diameter of 0 mm to 20 mm, more preferably 5 mm to 15 mm.

10. 10. The method according to any one of claims 1 to 9, wherein the at least two fragments or the at least two members are fixed to the at least one support element in a substantially parallel position relative to one another.

11. 11. The method according to any one of claims 1 to 10, wherein the at least two pieces are fixed to the at least one support element so as to be stacked on top of each other.

12. 12. The method of claim 1, wherein the at least two recruits of precious corals belonging to the Corallinae family are less than six months old.

13. 13. The method of claim 1, wherein each of the at least two recruits of a precious coral belonging to the Corallinae family is attached or affixed to an attachment substrate or growth substrate.

14. The method comprises: a') preparing at least two recruits of an existing precious coral belonging to the Corallinaceae family, each of which is attached or attached to an attachment substrate or growth substrate; b') fixing the attachment substrate or the growth substrate, respectively, to at least one support element so that the minimum distance between at least one of the accessions attached or added to the attachment substrate or the growth substrate, respectively, and at least one of the accessions attached to another attachment substrate or the growth substrate, is 0 mm to 5 mm; c) culturing the substrate fixed in step b') in seawater at a temperature ranging from 2°C to 30°C, preferably from 16°C to 22°C; 14. The method of any one of claims 1 to 13, comprising:

15. The method comprises: a') attaching or adding at least two juveniles of the pre-existing precious coral belonging to the Corallinae family to an attachment substrate or a growth substrate so as to obtain the at least two recruits; b') cutting the attachment substrate or the growth substrate to obtain a plurality of separate substrates, each having at least one of the accessions attached or attached thereto; b'') fixing each of the plurality of substrates to at least one support element so that the minimum distance between each of the added particles attached or added to each of the plurality of substrates and at least one added particle attached to another substrate of the plurality of substrates is 0 mm to 5 mm; c) culturing the plurality of substrates fixed in step b'') in seawater at a temperature ranging from 2°C to 30°C, preferably from 16°C to 22°C; 14. The method of any one of claims 1 to 13, comprising:

16. 10. The method of claim 9, wherein step b'') is performed such that a peripheral edge of each of the plurality of substrates contacts a peripheral edge of at least one other substrate of the plurality of substrates.

17. 17. The method of any one of claims 13 to 16, wherein the attachment substrate or the growth substrate is a sheet comprising or consisting of a polymeric material, preferably comprising or consisting of polyvinyl chloride (PVC), more preferably comprising or consisting of PVC.

18. 18. The method of any one of claims 1 to 17, wherein the at least one support element is made from a material selected from concrete, ceramic, marble, plastic, or a combination thereof, or the at least one support element consists of nylon thread, preferably the at least one support element is made from type 2 Portland cement mixed with sand and water.

19. 19. A method according to any one of claims 1 to 18, wherein the at least one support element comprises or consists of a seabed element, preferably selected from carbonate rock, coral rock, the remains of a precious coral colony, or a combination thereof.

20. 20. The method according to any one of claims 1 to 19, wherein the fragments or the members are fixed to the at least one support element so as to form a predetermined pattern, in particular the pattern is linear, striped, beam-shaped, triangular, square or circular.

21. 21. The method according to any one of claims 1 to 20, wherein step a) comprises at least one step of cutting off an existing colony of precious coral to obtain the at least two fragments.

22. Step c) comprises: open sea, A controlled marine environment selected from natural or artificial bodies of water, tanks, or aquaria 22. The method of any one of claims 1 to 21, comprising transplanting the cell culture medium into a host environment selected from the group consisting of:

23. 10. The method of claim 9, wherein the at least one support element is positioned in the open sea at a depth below the summer thermocline of the open sea, preferably in the depth range of 15 m to 2000 m, more preferably 15 m to 60 m.

24. 24. The method of any one of claims 1 to 23, wherein the at least one support element is fixed to an apparatus suitable for culturing corals in the sea or in a coral nursery.

25. 25. A method according to any one of the preceding claims, wherein the at least two fragments are chimeras of precious corals belonging to the family Corallinae, and the chimeras are generated from at least two recruits of precious corals using a method according to any one of claims 1 to 24.

26. A method for culturing precious corals, or colonies, recruits, or chimeras of precious corals, comprising: a) providing at least two fragments or recruits of existing precious corals belonging to the Corallinaceae family; b) fixing said fragments or said members to at least one support element so that the minimum distance between each of said fragments or members and at least one other of said fragments or members is between 0 cm and 2 cm, preferably between 0 mm and 5 mm; c) culturing the fragments or the accessions fixed in step b) in seawater at a temperature ranging from 2°C to 30°C, preferably from 16°C to 22°C; A method comprising:

27. 26. The method of the preceding claim, wherein the fragment or the participant is as defined in the method of any one of claims 1 to 25.

28. 28. The method of claim 26 or 27, comprising the steps of the method of any one of claims 1 to 25.

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