IMPROVED METHOD OF PROCESSING SEA CUCUMBERS, WITH NEW COMPOSITIONS

OF MATTER

BACKGROUND OF THE INVENTION 1. Field of the Invention

The present invention relates generally to methods of processing sea cucumbers and, more particularly, to improved sea cucumber processing methods are disclosed that minimize water use, improve air, water and land contamination, produce marketable products, and produce distilled water as a by product .

2. Description of the Related Art

Sea cucumbers are processed world-wide for their body-wall meat, and for their longitudinal muscle meat, as well as occasionally for production of fermented parts of the intestine, especially the gonadal tissue, referred to as Konowata in Japan. Processing, in most instances, involves manual cutting open of the body wall of the animal, excision of the guts, excision of the muscle meat either manually or by machine. The muscle meat is usually frozen, and the body wall is usually boiled and dried for the Asian market. For Cucumaria frondosa, the traditional processing includes manual cutting and draining of the intestinal Coelemic fluids into fresh water flumes that carry that same water to discharge overboard into the sea. Likewise, the gut contents and top and/or bottom pieces of the animal are cut and scraped manually. These portions of the animal are then ^' discarded into the same fresh water flume whereby those solid pieces must eventually be separated before ocean discharge, with the water passing as treated or untreated into the sea.

It is a further problem within the Cucumaria frondosa processing industry, that certain flesh contact and air-borne micro-particulate matter associated with cutting and scraping the animal, brings mucosal reactivity to certain individuals,

wherein such sensitive individuals react to the processing environment with excessive nasal discharge, inflamed ocular tissues, pulmonary discharge, rash on contact areas such as the hands, wrists and forearms. In the Cucumaria frondosa processing industry as practiced in the US and Canada, the skin with muscle intact is clamped to an angled scraping board after initial cutting of the body wall, spilling of the coelemic fluids, cutting off of the anterior and/or posterior of the animal and removal of gut contents. Once clamped onto the cutting board, small pieces of gut content are brushed with a bristle brush under a continually running small stream of fresh water. That 'meat-washing-water' is directed then to a fresh-water trough or flume where it is eventually screened of solids and dumped either treated or untreated into the sea.

The Cucumaria frondosa processing industry also involves the cooking of the skins of the animal prior to drying. Since water weight of the animal is decreased and adds to the cooking water volume, there is continual boiling over of fresh cooking water into drains commonly used for all discharge wastes. Since this boiling water contains potentially fish-toxic glycosides, this water should not be considered benign to the environment, and should be contained or commercially utilized in some manner. Either at the end of the processing day or throughout the day, the water is drained and/or re-filled.

This processing methodology presents several problems such as :

1. huge amounts of water usage for transport of the un-wanted portions of the animal;

2. further need for additional separations of the solids from the transport water prior to dumping into the sea or land-fill operations;

3. the risk of fish toxicity of the treated or untreated processing and cooking water;

4. physical reactions to those sensitive individuals from aerosolized or dermal contacted sea cucumber processing moisture, or fluids;

5. decreased ease of recovering marketable products from the water carried waste processing material such as the top and bottom piece of the animal or the gut content; and

6. unnecessary expense of handling the wastes after unnecessary contamination of that waste during the processing of commercial products.

Therefore, there exists a pressing need to improve the sea cucumber processing industry that is fraught with the above mentioned problems of waste disposal, clean water environmental fish health concerns, and human worker health during processing. There also exists the need to recoup expenses and to improve profitability by adding 'value-added' products to the processing efforts.

Yet other problems exist associated with Airborne contaminants. Since there exists a potential health hazard to workers from processing of sea cucumbers; such hazard presumed to be emanating from airborne moisture-laden • particulate matter, a method of worker protection is disclosed that minimizes said exposure risk by means of a simple air-removal ducting system that is preferentially positioned near the worker cutting area where initial sea cucumber aerosolized pathogens may occur. This vacuum air-flow ducting is divided under or over the worker area, the intake suction of which is near the source of origin of aerosolized contamination of breathing or skin contact air, and is ducted though hydrophilic filters and/or heat exchangers to prevent undue loss of building heat, and finally ducted outdoors, such process of ventilation is known to those skilled in the the 'ventilation' sciences, but is a novel step brought to bear upon the aerosolized contaminants plagueing the sea cucumber

-A- processing industry presently.

SUMMARY OF THE INVENTION

The field of this invention is the sea cucumber processing industry wherein processing and waste disposal challenges hamper economical net profit return on investment dollars. In the 'orange-footed-sea cucumber,' Cucumaria frondosa processing as it is currently employed, the waste disposal represents a considerable expense, despite patents describing the novel use of some of the particular wastes (Collin patents) . Additionally, the intestinal wastes, if contacted extensively with water that will eventually be discharged into bodies of water such as the sea or fresh-water streams, ponds or lakes, presents a certain amount of 'fish toxicity ¹ inasmuch as there exist toxins in most sea cucumbers that increase mortality in fish. Other processing chemicals -known to those skilled in the sewage remediation arts, pose environmental problems from overboard discharge outlets to public waters. It is an object of the present invention to provide novel methods to utilize hertofore discarded wastes, and compositions of matter resulting from such novel methods as well as elucidating the general utility of such compositions of matter. It is an additional object of the present invention to reduce water usage and remediation at sea cucumber processing plants, while at the same time reducing air-borne contaminants deleterious to human health. The present sea cucumber processing technology utilizes copious amounts of water; discards millions of pounds of potentially marketable products, and puts at risk, workers health, nearby environmentally sensitive bodies of water and land used for dumping such wastes.

An object of the present invention is an improved method for processing Cucumaria frondosa or other sea cucumber species, that utilizes an industrial vacuum system for immediate removal ^■ and storage of intestinal material from the

animal .

It is an object of the present invention to provide a means of removing toxic sea cucumber compounds from any over-board ocean discharge wastes, while at the same time, improving the methods of isolating certain materials for commercial ends.

Because fish processing always involves production of various kinds of organic contaminations to a water waste stream, and because such waster waste stream is in many cases, discharged over-board to the sea, there is a pressing need to purify said discharge water after any processing contamination. This, in many instances, is common technology in the fin-fish processing arts. It is not, however, known to the sea cucumber processing arts as currently practiced. Additionally, fin-fish processing, crustacean processing and other types of fish processing do not produce glycoside materials that need to be isolated for commercial ends, but also because of the need to keep such glycosides from potential for harm to fish environments where such discharges are occurring.

Therefore it is an object of the present invention to provide a means of sequestering sea cucumber glycosides from an aqueous sea cucumber processing waste stream.

An additional method is disclosed that utilizes a macerating pump prior to solids separations from intestinal mass, in which permeate through a screen produces an aqueous intestinal mass and retentate from screening produces solids for further processing. Said screening of sea cucumber processing wastes can also aid in pumping of said wastes to tanker trucks or other storage and removal equipment. In a further embodiment of the present invention, a method is disclosed describing a filtering and evaporation process for reducing cooking water wastes for later use in biomedical industries, and as food additives. A method is described for production of usable protein tissue high in beneficial lipids,

produced by screening, evaporation of inherent moisture and grinding.

It is a further object of the present invention to provide a composition of matter produced from sea cucumber intestinal mass, and method to produce same.

A method of production is disclosed to produce a lipid-bearing protein powder from sea cucumber intestinal material. The product by process has usefulness in the nutritional health sector and is raw material for further processing steps.

DESCRIPTION OF THE PREFERRED EMBODIMENTS This present invention is an improvement of traditional methods and is referred to as Vacuum, Evaporation and

Filtration (VEF Method) . This new method improves upon all of the parameters listed above and allows for further processing and merchandising of what previously was waste materials and/or the safe discard of any waste without adversely impacting the environment.

It has been surprisingly found by the inventor, and is an aspect of the present invention, that sea cucumber coelemic fluid can be isolated from any discharge processing water by a simple 'cutting grate' which allows the coelemic fluids of the animal to spill through the grate to a piped catchment apparatus that can then be clarified by known methods, and subsequently filtered by micro, ultra, nanofiltration and then reverse osmosis to obtain a novel biologically active sea cucumber fraction for further commercial exploitation or reduction under reduced pressure to form a powder. Previously, no standard process for obtaining coelemic fluids during processing was in existence, nor was there any potential use for such coelemic fluids other than for laboratory use only. Thus the specifications of the present invention address a need in the sea cucumber processing industry.

Coelemic fluids are oftentimes found coming from the fish 'totes' in which the animals are transported from the fishing boats. This fluid can accumulate in the chilled holding rooms, in the transportation truck or wherever the animals are stored. Since this fluid contains glycosides of potential medical interest, it is an object of the present invention to provide a means of sequestering these ' waste-coelemic fluids' for future use or merely for safe disposal. Specifically, any source of said sea cucumber coelemic fluids can be identified and said liquid can be pumped to evaporation tanks after traditional clarification of solids, as is commonly done in the waste-water processing industry. The resulting wastes, in a reduced form as a powder or thick salty liquid can be further utilized or when in a liquid form, can be ultra and nanofiltered and finally subjected to reverse osmosis, as is commonly utilized in other industries to form raw materials for medical research or as crude drugs of a particular molecular weight sized class. It is known that sea cucumber coelemic fluids contain certain biologically active cytokines and cytokine inhibitors of various molecular weights. It is an object of the present invention to provide a novel means of producing such raw material at an industrial quantity.

In the processing room, near said 'cutting grate' can be attached a vacuum system that can be activated from sensors or manually or can be run non-stop whereby the gut contents can be vacuumed up and discharged into holding tanks or bins, as was widely done in the US poultry industry as a means of quickly removing the poultry entrails from the meat processing environment . It is an additional aspect of the present invention that the 'meat-washing-water' can be minimized by employment of motion or other sensors on the clamping devise or other sporadically actioned device, wherein activation manually of the clamping or other device begins the ^' water flow during the 'meat-brushing' operation. This improvement decreases water

usage considerably and decreases the amount of water that needs to be treated or discharged. The brush itself can contain a vacuum ou'tlet that can discharge those 'meat-washing-wastes' to the same vessel or bin as the gut content. In another embodiment of the same aspect of this invention, such 'washing water' can be recycled after clarification, thus reducing the water usage, and the volume of water needed to be treated prior to the waste management component of the industrial process. New methods to reduce water consumption and obtain usable product. Cook-water from cooking pots is drained or pumped to clarifiers as are known to persons skilled in the waste-water processing arts, and then to a series of steam-jacketed evaporation tanks that are under vacuum of approximately 20 inches of mercury. The cook-water so processed is relieved of between 50 and 75% of its water weight, removed from the tank as a sludge and further dried by any conventional means, including freeze-drying, rotary vacuum drying, spray drying or fluid-bed drying to produce a powder. This process removes the potential for environmental harm occasioned by cooking water discharge into a waste stream destined for overboard discharge into the sea. It is also a novel method to sequester a raw material for subsequent production of sea cucumber glycosides, lipids and proteins. The cook-water can also be subjected to various filtration processes at any step during its aqueous evaporation process, as might be convenient, depending upon the constraints of any filtration process employed

Additionally, the present invention is intended to provide a means of sequestering sea cucumber glycosides from an aqueous sea cucumber processing waste stream. By way of example, and not as a limitation, the following examples are indicative of the improvements to be realized from the present invention: 1. Air exchange improvement. In a sea cucumber

processing plant, an air removal system is placed in a manifold fashion, with suction outlets positioned near each worker. PVC 4 inch piping, with caps or valving of said piping accommodates the need to take unused worker position suctions off-line if desired. The 4 inch line is connected to forced air suctions sufficient to produce a fume or smoke suction at the worker site of at least two feet of 10 cubic feet per minute. Said line is capable of quick cleaning during plant-clean-up operations by anti-microbial solution washing, ease of take-down, water flooding and the like.

2. "Cutting board improvement." It is an object of the present invention to provide a process for industrial isolation of sea cucumber coelemic fluids, both with a goal of decreasing contamination and also for the goal of production of a novel raw material for further processing of said coelemic fluids . Worker cutting positions are equipped with a positionable cutting board with a slight containment aspect of a rim around a cutting area of between 3 and 8 inches that has as its base, a plastic removable and cleanable disk with drilled or punched holes, or other means of liquid drainage leading to piping or vessel containment beneath the table. In the case of piping, all processing stations on the processing line are equipped with connectivity to a central vessel that can be further pumped to evaporation and/or filtering processes. In the case of each station having its own vessel, such vessel can be as simple as a plastic 5 gallon bucket or the like, which is emptied as necessary. Prior to entry into a piping system or bucket, a removable and cleanable screen can be placed that grossly filters gut pieces from the coelemic liquid being saved.

3. Vacuum waste removal. A vacuum pump suitable to create sufficient vacuum in as many worker cutting stations as necessary is attached through cleanable piping to vessels which can contain the sea cucumber processing wastes, as are known to those skilled in the poultry processing arts. The

work stations are equipped with suction ends connected to the vacuum system either as flexible wands or as stationary suction ends near the cutting boards. Workers lift -the sea cucumber skin with guts up to the vacuum inlet, or apply the suction end via a flexible hose to the skin area of the animal, or with the cutting knife, scrape the loosened guts towards the vacuum opening, or if cutting is mechanized, contact the vacuum opening to the guts as possible depending upon the configuration of the mechanized operation, bring the vacuum system into contact with the gut wastes or products. 4. Direct waste removal without vacuum. In another embodiment of the present invention, the scraped guts are not contacted with water after removal, but are directed to vessels or bins that' do not need the transport properties of the flowing water, as is commonly used. This protects the water from future contamination such as with ichthyotoxins and increased levels of proteins creating biological oxygen demand and related contaminations resulting from processing contact of water with gut contents. 5. Liquifaction of gut contents. Once removed from the vacuum or non-vacuum holding vessel or containment apparatus, or while still within such vessel or apparatus, the gut contents are churned, agitated or otherwise partly macerated with pumps known to those in the sewage processing arts, which have as an object the liquefaction of the highly aqueous gut contents which is approximately 90% water as it comes from the sea cucumber. Because the scrapings of guts may also contain the removed top portion of the animal known as the tentacles or 'flower end' and referred to as T-2000 in the patents of Collin, or the removed bottom cap hereinafter referred to as

'caps,' this agitation separates such semi-solid portions from any attached aqueous gut tissue. Once this separation occurs, then the slurry is pumped to separation devices as are known to those in the 'de-watering' arts, and the solids are directed towards separate holding vessels, bins, or the like.

Liquification can also occur by simple grinding of semi-solids into the aqueous gut material as described above, or with a mincing machine as is commonly known in the fishery arts.

6. Mechanical separation of oils, water and sediments from sea cucumber processing. From the catchment basins that retain macerated gut and or gut mixed with other tissue portions of the sea cucumber, the slurry is pumped to round stainless tanks of approximately 500 to 1000 gallon size that are equipped with a motorized apparatus situated in the center of the tank, whereby a shallow draft arm may skim the uppermost layer from the said holding tank at an angle that leads or pushes the uppermost layer to a separate holding tank, such apparatus being referred to as an 'oil skimmer' as is known in the sardine processing arts. The so called "oil skimmer' may be, in fact, any apparatus that can effectively 'skim' the uppermost layer of oils or flotsam from the holding tank containing the ground-up sea cucumber tissues.

Oil and flotsam debris obtained in this manner is further subjected to filtering of various size mesh screens to remove any foreign material, and is then brought by pipes or flumes to additional centrifuges and clarifiers as are known to the sewage processing arts, that may further separate remaining contaminant/aqueous materials. Oils and oily material thus obtained are piped to oil containers of 20 to 55gallon size, are treated with anti-oxidants such as tartaric acids, or mixed tocopherols, or green tea catechins, or any other suitable anti-oxidant known to the oil-processing arts and are ready for market or further processing.

After oil skimming, the remaining water and tissue sediments are further screened and centrifuged to separate these two additional layers, an aqueous pigmented layer, and a sedimentary tissue layer.

Aqueous pigmented layer.

6. Reduction of aqueous slurry wastes and preparation of dried gut wastes. Once the gut slurry is separated from the semi-solids the aqueous slurry is directed to evaporation vessels as are commonly known to those skilled in the evaporation arts. Or, if semi-solids are desired to remain in the finished slurry, the entire gut contents as it came from the cutting operation is directed to the evaporation tanks in any manner possible. Within the vacuum evaporation tanks, vacuum is created sufficient to reduce the evaporation temperature, and the moisture content of the slurry is reduced to a percent necessary and appropriate for the next operation, and is referred to as 'reduced wastes.' 7. Reduced waste drying process. Such reduced wastes as described above are then produced into additional useful products, the steps of which can include but are not limited to: a. Rotary drum, tray, heat-pump, air or other types of industrial drying to produce an approximate 10% moisture dry product b. Freeze-drying to produce a 1 -8% dry product c. Tray drying to produce a dry product d. Centrifugation /separation of gut tube wall away from gut contents by common methods which may include cell-wall disruption, grinding, liquefaction and the like. e. Acidification of wastes to stabilize microbial degradation f. Acidification of reduced wastes with water content appropriate for protein precipitation with subsequent additional steps comprising but not limited to: i. Application of dissolved air and filtration as is commonly used in sewage treatment

plants ii. Floation or precipitation of proteinaceous/lipidic material away from aqueous material, iii. Removal of products of precipitation by common methods iv. Filtration of aqueous portion of acidification through micro, ultra, and nanofiltration as is commonly used in the filtration arts, with final reverse osmosis filtration to separate target compounds or to simply remove remaining salts. v. Classification of the products of the various filtration steps according to molecular weight cut-off filters utilized and desired target material, which may include, but is not limited to:

1. separation of lipids and phospholipids away from non-lipid material

2. separation of Glycolipids and saponins by use of filtration processes g. enzyme hydrolysis of any step above to hydrolyze the gut content proteins wherein the so hydrolyzed slurry is ready for any future product development step such as molecular weight cut-off filtration, centrifugation, clarification, classification, and the like, depending upon the target products so desired.

8. Cook-water development. Cook-water (CW) is first screened by various sized screens and then clarified as per methods known in the sewage processing arts, (with solids being removed and disposed of appropriately) and the clarified water is then directed to vacuum evaporation vessels for reduction of the water content to a desired percentage, with optional additional steps including but not limited to:

a. Micro-ultra and nanofiltration with reverse osomosis to remove protein and/or lipidic compounds for commercial ends i. Specifically, microfiltration is utilized on crude clarified cook-water to remove lipids and proteins entrained in the water, which are sequestered as retentate ii . Specifically, ultra-filtration uses as starting material the permeate from previous step of micro-filtration, with molecular weight cut-offs of between 2,000 and 30,000 to remove as retentates, proteins or peptides of commercial value, and especially a filtration step consisting of passing the permeate from a 2,000 mw filter through a filter of between 500mw and 1,500 approximate mw to obtain glycosides in the range of 800 to 1400mw. iii . It is an object of the present invention to provide a means of cleaning sea cucumber processing water that has been contaminated with fish toxins related to glycosides. Passing such cook-water and process water through micro, ultra and nano-filtrations filter systems produces a relatively toxin-free eluate that is acceptable to most regulatory agencies having oversight of aqueous discharge to the environment. b. Acidification of the aqueous CW aimed at separation/precipitation of proteins away from other compounds c. Acidification of the aqueous CW aimed at separation/precipitation of lipidic materials away from more polar compounds d. Spray drying of the aqueous material from any point of classification or filtration step, or prior to any such step or reduction of water content e. Separation of saponins from the cook-water by use of

ultra-filtration molecular weight cut-off filter separation in the range of 1000 to 2000 mw.

9. Saponin Development. Dry Cook-water or any product resulting from the CW processing steps listed above, or additional steps that are evident to any person skilled in the saponin isolation arts, is contacted with a non-polar de-lipidizing solvent such as supercritical CO2, non-polar organic solvent as is commonly known to those in the deplipidizing arts, to remove non-polar materials from the CW or CW sub-fraction as described herein. The de-lipidized material is then contacted with supercritical C02/ co-solvents (70-30) of ethanol/water (85-15) to produce an extract that is then subjected to ultra and nanofiltration to sequester any compound between the molecular weights of 1000 and 2000, as a crude saponin mixture. Optionally, the saponin-bearing extract can be complexed with cholesterol in ethanol or other liquid solvent, with additional steps of organic solvent or supercritical CO2 separation of the cholesterol from the complexed polar saponin, the steps of which may include: a. Addition of saponin bearing extract from above to cholesterol (1:15) in an organic solvent, with agitation at room temperatures for 5 hours. b. Evaporation of said solvent to produce a complex of saponins and cholesterol c. Separation and isolation of saponins from cholesterols by removal of cholesterol by non-polar solvents as are known to the chemistry arts.

10. Peptide production from sea cucumber processing wastes. The Collin patent provides a novel means of producing anti-inflammatory compounds by means of utilizing a 'de-boner/mincer, and sequestering the outer epithelial layer away from the inner endothelial layer. The same method is used to produce a non-bitter protein product of the gut

wastes. The methods of which include, but are not limited to: a. Drying of the gut wastes by any means, as described in the U.S. Patent No. 6,055,936 and 6,399,105, both issued to Collin, and incorporated herein by reference as if fully rewritten herein. b. Removal of non-polar lipids, as described in the U.S. Patent No. 6,055,936 and 6,399,105, both issued to Collin, and incorporated herein by reference as if fully rewritten herein. c. Obtaining of a ' de-lipidized ¹ gut material, which still contains polar lipids such as phosphatidylcholine, and other phospholipids classes, including Glycolipids d. Supercritical CO2 plus-co-solvent extraction of the de-lipidized material, or by polar solvents such as water/alcohol, alcohol, serial extraction by acetone followed by alcohol, or water/alcohol solvents, removal of all solvents by traditional means, and sequestering the products of said solvent process as 'polar lipids' and lecithins, suitable for future processing as is known in the lecithin processing arts, leaving a light tan power or granular material that is thus more or less completely de-lipidized and suitable for inclusion as a food ingredient. e. The 'protein powder of the previous step' is then mixed with water and hydrolyzed enzymatically by any means suitable and as known to those skilled in the protein hyrolysate arts, and filtered through a 10,000 molecular weight cut-off filtration step. This step produces as a permeate, a white peptide material if spray dried after this step. Additionally, if in its aqueous 10,000 mw permeate condition it is further filtered through a 2,000mw filter, there will remain as a retentate, a mild-tasting white peptide powder which can be produced through spray drying. The permeate from this step contains glycosides and salts and can be the raw

material of a glycoside-bearing commercial material used as is, or as a feed-stock for further purification of any glycoside contained therein. Through further filtering of this 2,000mw permeate with a 500mw nano-filter, salts can be removed, thus further purifying the glycoside material. The above disclosures do not mean to be limiting in their steps and any person skilled in the filtration arts can discern additional steps and implications, without adding novelty to the process. f. Production of lipids, glycosides and proteins from processing waste. In any fishery waste system that utilizes the separation of proteins away from the aqueous ^ι components of the waste stream, such as what is known as a 'Disolved Air Filtration' system, additional steps to utilize commercial products from that protein/lipid separation step can be instituted that include but are not limited to the steps of: i. Sequestering said protein-bearing phase by means known to the sewage-treatment arts ii . Drying said protein-bearing material through any known means available iii. Solvent or supercritical or sub-critical co2 or other gas contacting of the material in its dry or moist condition to selectively remove lipids away from proteins, glycosides away from proteins and/or lipids, phosphorus-bearing compounds away from non-phosphorous-bearing compounds, as are known to those skilled in the solvent or supercritical or sub-critical co2 extraction arts. The foregoing descriptions of specific embodiments of the present invention have been presented for purposes of illustration and description. They are not intended to be ^■ exhaustive or to limit the invention to the precise forms disclosed, and obviously many modifications and variations are possible in light of the above teaching. The embodiments were

chosen and described in order to best explain the principles of the invention and its practical application, to thereby enable others skilled in the art to best utilize the invention and various embodiments with various modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the Claims appended hereto and their equivalents. Therefore, the scope of the invention is to be limited only by the following claims.