JP5680413B2 - Survival limit temperature tolerance imparting agent for seafood and method for culturing seafood - Google Patents

Description

  [Technical Field] The present invention relates to a fish or crustacean survivability limit temperature tolerance-imparting agent containing arginine and a method for culturing fish or crustacea characterized by feeding arginine.

  Fish and crustaceans are thermophilic animals, so they are less susceptible to changes in environmental temperature than thermostatic animals, and have biological characteristics that cause death if the water temperature deviates from the individual's survival limit. Due to the arrival of abnormal weather phenomena such as global warming in recent years, the temperature of aquaculture ponds where fish or crustaceans are cultivated often exceeds 29-30 ° C. Mass death of fish or crustaceans to be cultured Is a problem. On the other hand, in the winter season, the temperature of the pond is 7 ° C. or less, causing a large amount of death of fish or crustaceans to be cultured.

As a method for reducing the stress of fish derived from the cultivation of crustaceans and short-term temperature changes, a method of feeding a feed containing DHA or its ester and / or phospholipid is known (Patent Document 1). ).
JP-A-9-121784

  However, there are no additives and aquaculture methods that allow fish or crustaceans to withstand rearing at abnormal water temperatures for longer periods, and their development is desired.

  It is an object of the present invention to provide a novel formulation that confer fish or crustacean temperature marginal survival and to use the formulation at a water temperature that exceeds the unsuitable temperature of fish or crustaceans, i.e. It is another object of the present invention to provide an aquaculture method in which fish or crustaceans can survive.

  As a result of diligent efforts, the present inventors have found that when arginine is added to the feed and fish and crustaceans are bred, fish or crustaceans can be bred even in water temperatures where these marine animals are not viable. I found. Based on this finding, the present inventors have completed the present invention.

  In addition, the method of adding an amino acid to a feed as a nutrient for seafood is known (Japanese Patent Laid-Open No. 2001-231461), and the optimum content of arginine or tryptophan for the growth of prawns is in each feed. It is known to be 3.0% or 1.0% (Akio Kanazawa: Shrimp and Crab Augmented Culture (edited by Tachibana Takajiro et al.), Hoshiseisha Koseikaku, Tokyo, 1996, 226-250). A method of adding tryptophan or a derivative thereof to the feed to prevent cannibalism of the crustaceans (JP-A-5-7463, JP-A-6-78687), and arginine 3.5 mmol / Kg together with alanine to promote the growth of fish There is known a method for promoting insulin secretion in fish by intramuscular injection (Japanese Patent Laid-Open No. 2007-49938). However, the pharmacological effect of arginine for imparting the limit of survival temperature tolerance to fish or crustaceans is not known, and was discovered for the first time by the present inventors.

  That is, the present invention relates to the following (1) to (25).

(1) A life limit temperature tolerance imparting agent for fish or crustaceans characterized by containing arginine.

(2) The fish or crustacean survival limit temperature tolerance-imparting agent according to (1), further comprising tryptophan.

(3) The fish is red sea bream, flounder, yellowtail, amberjack, tiger puffer, tuna, eel, sweetfish or rainbow trout, and the crustacean is prawn, bovine shrimp, white leg shrimp or crab (1) or (2 ) Fish or crustacean survival limit temperature tolerance imparting agent.

(4) The survival limit temperature tolerance-imparting agent of any one of (1) to (3), wherein the arginine content is 3.5% (w / w) or more.

(5) The survival limit temperature tolerance-imparting agent for crustaceans according to (4), wherein the arginine content is 3.5 to 5.0% (w / w).

(6) The survivor temperature tolerance imparting agent for crustaceans according to any one of (2) to (5), wherein the tryptophan content is 0.6% (w / w) or more.

(7) The survivor temperature limit imparting agent for crustaceans according to (6), wherein the tryptophan content is 1.6 to 2.0% (w / w).

(8) The survival limit temperature tolerance-imparting agent for fish according to any one of (1) to (3), wherein the arginine content is 3.0% (w / w) or more.

(9) The survival limit temperature tolerance-imparting agent for fish according to (8), wherein the arginine content is 3.0 to 5.0% (w / w).

(10) When the target is a crustacean, arginine is added to the aquatic feed so that the arginine content is 3.5% (w / w) or more, and the target is fish. In this case, a method for cultivating fish or shellfish, characterized by adding arginine to aquatic feed so that the arginine content is 3.0% (w / w) or more.

(11) A method for cultivating fish or crustaceans, comprising feeding the survival limit temperature tolerance-imparting agent for fish or crustaceans according to any one of (1) to (9).

(12) The fish is red sea bream, flounder, yellowtail, amberjack, tiger puffer, tuna, eel, sweetfish or rainbow trout, and the crustacean is prawn, bovine shrimp, white leg shrimp or crab (10) or (11 ) Fish or crustacean culture methods.

(13) The method for culturing fish or crustaceans according to any one of (10) to (12), wherein the water temperature of the farm is 29 ° C or higher or 12 ° C or lower.

(14) The method for culturing fish or crustaceans according to (13) when the water temperature of the farm is 29 to 33 ° C or 7 to 12 ° C.

(15) Fish or crustacean aquaculture characterized by feeding a feed containing 800 mg or more of arginine per kg body weight of fish or crustacean per day when the water temperature of the farm is 29 ° C or higher or 12 ° C or lower Method.

(16) The method for cultivating a fish or crustacean according to (15), further comprising feeding a feed containing 180 mg or more of tryptophan per kg body weight of the fish or crustacean per day.

(17) Use of arginine for producing a crustacean surviving limit temperature tolerance-imparting agent.

(18) Use of arginine according to (17), which is added to an aquatic feed so that the arginine content is 3.5% (w / w) or more.

(19) Use of arginine according to (18), which is added to an aquatic feed so that the arginine content is 3.5 to 5.0% (w / w).

(20) Use of arginine for producing an agent for imparting a survival limit temperature tolerance of fish.

(21) Use of arginine according to (20), which is added to an aquatic feed so that the arginine content is 3.0% (w / w) or more.

(22) Use of arginine according to (21), which is added to an aquatic feed so that the arginine content is 3.0 to 5.0% (w / w).

(23) Arginine for conferring survival temperature tolerance on fish or crustaceans.

(24) Feed to the crustaceans to be 3.5% (w / w) or more in the aquatic feed, or 3.0% (w / w) or more in the aquatic feed (23) Arginine to feed fish including.

(25) The arginine according to (23) or (24), wherein fish or crustaceans having a water temperature of 29 ° C. or higher or 12 ° C. or lower are fed with 800 mg or more per kg of body weight per day.

  This specification includes the contents described in the specification and / or drawings of Japanese Patent Application No. 2008-190853, which is the basis for the priority of the present application.

FIG. 1 shows the results of changes in survival rate of prawn prawns after high temperature load in the test group administered with the arginine-containing preparation and the control group administered with the comparative preparation. FIG. 2 shows the results of changes in survival rate of red sea bream after high temperature load in the test group administered with the arginine-containing preparation and the control group administered with the comparative preparation.

  Hereinafter, the present invention will be described in detail.

  The fish or crustacean survival limit temperature tolerance-imparting agent characterized by containing the arginine of the present invention is an agent that is administered at an inappropriate temperature by administering arginine in excess of the normal fish or crustacean nutrition. It is a marine product having an effect of reducing growth disorders or improving the survival rate at the survival limit temperature.

  In the present invention, the unsuitable temperature refers to a temperature range from the temperature at which fish or crustaceans develop a growth disorder to a lethality, that is, a temperature range from a survival limit temperature to a temperature range including the unsuitable temperature and the survival limit temperature. Usually, it is 7 to 12 ° C and 29 to 33 ° C although it varies depending on the species.

  Further, the fish or crustacean survival limit temperature tolerance-imparting agent characterized by containing arginine of the present invention may contain an effective amount of tryptophan in addition to arginine. Furthermore, generally, since the marine medicinal product and the feed additive are added to the feed and fed, they may contain components contained in a general marine feed pellet.

  The survival limit temperature tolerance imparting agent for fish or crustaceans containing arginine according to the present invention may be any one that contains arginine and can be consumed by fish or crustaceans. Arginine preparations that are specially prepared for addition to feed prior to feeding, etc. can be mentioned, but in order to efficiently express the pharmacological effects of arginine, arginine is added to fishery feed. It is preferable that they are formed.

  The aquatic feed may be any feed as long as it is used as a fish or shellfish aquatic feed. For example, aquatic feed pellets such as dry pellets and moist pellets may be used. Is preferably used. As the aquatic feed pellet, a commercially available product or a product prepared by blending components commonly used in aquatic feed by a conventional method can be used.

  The aquatic feed is an aquatic feed composed of natural ingredients, erythromycin preparation, ampicillin preparation, praziquantel preparation, lysozyme chloride preparation, oxytetracycline hydrochloride preparation, spiramycin preparation, sodium niflustyrene ester preparation, lincomycin hydrochloride preparation, flumequin. Pharmaceuticals, marine products such as glutathione preparations, vitamins such as vitamin C, vitamin B1, vitamin A, vitamin D and vitamin E, nutritional supplements such as amino acids such as lysine, methionine and histidine, β-carotene, astaxanthin, It may be an aquatic feed in which pigments such as canthaxanthin and the like (hereinafter, these substances are collectively referred to as aquatic additive substances) are mixed or spread.

  These aquatic additives are preferably contained in the feed in an amount of 0.01 to 10% (w / w), more preferably 1.0 to 5.0% (w / w). Examples of other components contained in the fishery feed include crude protein, crude fat, crude fiber, and crude ash.

  When arginine is mixed with aquatic feeds and used as a crustacean survival limit temperature-imparting agent, the final arginine is 3.5% (w / w) or more in combination with the arginine normally contained in crustacean feeds. It may be contained, but since growth failure may occur at a high concentration, arginine is preferably 3.5 to 7.0% (w / w), particularly preferably 3.5 to 5.0% (w / W) may be contained.

  Furthermore, when tryptophan is mixed with the arginine-containing aquatic feed, tryptophan may be finally added in an amount of 0.6% (w / w) or more, preferably 1.6% (w / w) or more. In this case, the upper limit of the content is preferably set to 3.0% or less from the viewpoint of growth inhibition, and particularly preferably 1.6 to 2.0% (w / w) of tryptophan may be contained.

  In addition, 2.8-3.0% (w / w) of arginine derived from natural raw materials and about 0.5-1.0% (w / w) of tryptophan are pre-existing nutrients in crustacean marine feed In this case, the necessary amounts of arginine and tryptophan obtained by subtracting the contents of arginine and tryptophan derived from natural raw materials in the feed from the final concentration may be added.

  Therefore, in the present invention, the content of the shellfish feed including arginine derived from a natural raw material is 3.5% (w / w) or more, preferably 3.5 to 7.0% (w / w), Particularly preferably, arginine is 0.5 to 0.7% (w / w) or more, preferably 0.5 to 4.2% (w / w) to be 3.5 to 5.0% (w / w). ), Particularly preferably 0.5 to 2.2% (w / w) may be required to be added. Further, if necessary, the content of the shellfish feed including tryptophan derived from natural raw materials is 0.6% (w / w) or more, preferably 1.6 to 2.0% (w / w), Tryptophan may be added in an amount of 0.1% (w / w) or more, preferably 0.6 to 1.5% (w / w).

  When arginine is mixed with aquatic feed and used as an agent for providing a survival limit temperature for fish, the final arginine is 3.0% (w / w) together with the arginine derived from natural ingredients contained in the fish feed. Although it is sufficient to contain the above, since a growth disorder may occur at a high concentration, arginine is 3.0 to 7.0% (w / w), particularly preferably 3.0 to 5.0% (w / w). w) may be contained.

  In addition, about 1.7 to 2.6% (w / w) of natural raw material-derived arginine may exist in advance as a nutrient in fish feed, but in that case, the content in the feed from the above final concentration What is necessary is just to add arginine which remove | eliminated each.

  Therefore, in the present invention, the content of arginine derived from natural raw materials in fish feed is preferably 3.0 to 7.0% (w / w), particularly preferably 3.0 to 5.0% ( w / w) It may be necessary to add arginine, preferably 0.4 to 5.3% (w / w), particularly preferably 0.4 to 3.3% (w / w).

  The method for producing a fish or crustacean survival limit temperature tolerance-imparting agent characterized by containing arginine may be prepared by preparing a feed preparation raw material prepared in advance so as to contain the desired amount of arginine as a pellet, The desired amount of arginine may be added to the pellets for aquatic feed.

  As a method for producing a feed preparation raw material prepared in advance so as to contain the desired amount of arginine as a pellet, any method may be used as long as the desired amount of arginine is contained. Manufacturing is mentioned.

  Arginine is 3.5% (w / w) or more in the case of feed pellets for crustaceans, and 3.0% or more in the case of fish pellets for fish. Addition of tryptophan and fishery products is necessary. To the feed raw material prepared so that the desired amount of the substance is blended, an oil and fat mixed with at least 3 to 10% of oil and fat such as fish oil, lecithin and sterols is added to the feed raw material, and If necessary, an aqueous solvent selected from water, hydrous ethanol and the like is added and mixed in an amount of 10 to 40% based on the total amount of raw materials, and then a survival limit temperature resistance-imparting agent is produced by an extrusion granulator and a fluidized bed granulator.

  Moreover, there is no restriction | limiting in particular in the method of spreading arginine on the commercially available feed pellet for fisheries, For example, the following methods can be mention | raise | lifted. That is, preferably 0.1 to 20% (v / w), more preferably 1.0 to 10% (v / w) water is placed in a tray or the like with respect to the fishery feed pellets, and the pellets are placed therein. The feed pellets are hydrated by a method of dipping and dipping, or a method of spraying water using a spray or the like.

  Next, the final concentration in the case of shellfish feed pellets is 3.5% (w / w) or more, and in the case of fish feed pellets, the final concentration is 3.0% relative to the weight of the fishery feed pellets. w / w) Above arginine, if necessary, add the desired amounts of tryptophan and aquatic additive and sprinkle the surface of the aquatic feed pellet as it is, or sprinkle using a sieve or the like.

  Alternatively, in the case of shellfish feed pellets in a suitable solvent such as water, buffer, ethanol, etc., the final concentration of the feed pellet weight is 3.5% (w / w) or more of the feed pellet weight for fish. In this case, arginine at a final concentration of 3.0% (w / w) or more is dissolved or suspended if necessary in the desired amount of tryptophan and aquatic additive, and the resulting solution or suspension is Arginine may be spread on the surface of the aquatic feed pellets by immersing the pellets or spraying the solution or suspension onto the aquatic feed pellets. The content of the fishery additive after spreading is preferably 0.01 to 10% (w / w), and 1.0 to 5.0% (w / w) with respect to the fishery feed pellets. More preferably.

  The survival limit temperature resistance-imparting agent processed into the pellet form may be coated with a coating agent in order to absorb arginine more efficiently.

  The coating agent used in the present invention may be any type, but it is preferable to use vegetable zein, and the coated survival limit temperature tolerance-imparting agent has a final concentration of zein of preferably 1.0 to 15%. (W / v), more preferably 2.0 to 10% (w / v), more preferably 4.0 to 8.0% (w / v), preferably 50 to 90% zein (V / v), more preferably 50 to 70% (v / v) can be prepared by dissolving in an aqueous alcohol solution.

  Twein can be prepared by a known method (for example, the method described in JP-B-07-84477, JP-B-07-25797, JP-A-05-30917, JP-A-61-167700).

  Moreover, you may use a twein (for example, Showa Sangyo Co., Ltd. product). Examples of the alcohol that dissolves twein include ethanol and edible alcohol, but it is preferable to use ethanol.

  If necessary, fatty acid or the like as a plasticizer may be added to the coating agent in an amount of 0.1 to 30% (w / v), preferably 1.0 to 10% (w / v). The fatty acid is preferably a fatty acid having high fusion activity with respect to cells, and examples thereof include oleic acid, linoleic acid, linolenic acid, icosapentaenoic acid, docosahexaenoic acid, and monoglycerides and lecithin composed of the above fatty acids that can be dissolved in an alcohol aqueous solution. Oleic acid is particularly preferably used.

  The method of coating with a coating agent may be any method as long as it is a method capable of coating aquatic feed pellets. For example, a method of immersing a fishery feed pellet in a coating agent or a method of coating a fishery feed pellet by spraying the coating agent with a spray or the like can be used.

  Specifically, the concentration of zein is preferably 1.0 to 15% (w / v), more preferably 2.0 to 10% (w / v), still more preferably 4.0 to 8.0%. Twein is preferably dissolved in an aqueous alcohol solution of 50 to 90% (v / v), more preferably 50 to 70% (v / v) so that the concentration becomes (w / v). The feed pellets for aquatic products are coated by the above immersion method or spraying method. After coating, it is allowed to stand at room temperature for 0.5 to 1 hour, whereby a coated aquatic feed pellet (hereinafter abbreviated as the aquatic feed of the present invention) can be obtained. It is preferable that content of the coating agent with respect to the survival limit temperature tolerance imparting agent of the present invention after coating is 1.0 to 10% (w / w), and 2.0 to 3.0% (w / w). More preferably.

  In the method for culturing fish or crustaceans according to the present invention, a feed containing 800 mg or more of arginine per kg body weight of fish or crustaceans per day is given to fish or crustaceans in order to confer fish or crustacean tolerance for survival temperature. It is characterized by feeding. In the aquaculture method, the water temperature of the aquaculture farm can be used at the unsuitable temperature or the survival limit temperature. The feeding period of this aquaculture method can be 1 to 4 months, preferably 2 months.

  The fish or crustacean culture method of the present invention may further feed the fish or crustacean with a feed containing 180 mg or more of tryptophan per 1 kg of fish or crustacean weight per day.

  Examples of fish that can be subjected to the cultivation method of the present invention include red sea bream, Japanese flounder, yellowtail, amberjack, tiger puffer, tuna, eel, sweetfish, rainbow trout, etc. Preferably, it is used for aquaculture of red sea bream, Japanese flounder, yellowtail, amberjack and prawn.

  As a feeding method of arginine in the aquaculture method of the present invention, the survival limit temperature tolerance-imparting agent processed into the aquatic feed pellets may be fed, but any method such as a method of adding arginine to the feed or a method of simply feeding arginine Such a method may be used.

  The aquaculture method of the present invention includes aquaculture that performs only seedling production (fry production) and aquaculture that performs production up to a parent fish.

  EXAMPLES Hereinafter, the present invention will be specifically described by way of examples, but these do not limit the scope of the present invention.

Example 1 Influence of Crustacea on Formulation Containing Arginine (I) Production of Formulation Containing 3.5% (w / w) Arginine

  Animal raw materials excluding fats and oils shown in Table 1 above, plant raw materials, and others (including 3.0% (w / w) arginine derived from natural raw materials, 0.6% (w / w) tryptophan) and arginine After 94.8 kg of the raw material component (0.5% (w / w)) was coarsely pulverized by a pulverizer, it was further finely pulverized by a fine pulverization classifier (manufactured by Hosokawa Micron).

  The obtained finely pulverized raw material was put into a blender (manufactured by Neotec), and further 5.2 kg of fats and oils described in Table 1 and 300 kg of water were added and mixed.

  The slurry produced by mixing was applied to an extrusion granulator (manufactured by Buehler), molded at a temperature of 60 ° C., and then dried at a temperature of 100 ° C. (manufactured by Nippon Dryer).

  The obtained pellets were sieved and the particle diameter was 0.2 mm. Arginine was increased by 0.5% (w / w) from the comparative preparation (containing 3.0% (w / w) arginine). % (W / w) -containing survival limit temperature tolerance-imparting agent (Formulation 1) was produced. In this example, the comparative preparation is a shrimp commercial feed manufactured by the same method as in Example 1 using a raw material obtained by removing only the amino acids arginine from the raw material composition shown in Table 1 above.

(II) Production of preparation containing arginine 4.0% (w / w)

  Using the raw materials shown in Table 2a (containing natural raw material-derived arginine 3.0% (w / w), tryptophan 0.6% (w / w)), a comparative preparation was prepared in the same manner as in Example 1. An arginine 4.0% (w / w) -containing survival limit temperature tolerance-imparting agent (formulation 2) was produced in which the amount of arginine was increased 1.0% (w / w) from the commercial feed.

(III) Production of 5.0% (w / w) arginine-containing preparation

  Comparative preparations were prepared in the same manner as in Example 1 using the raw materials shown in Table 2b (containing 3.0% (w / w) arginine derived from natural raw materials and 0.6% (w / w) tryptophan). Arginine 5.0% (w / w) increased survival limit temperature tolerance-imparting agent (formulation) in which arginine was increased by 2.0% (w / w) and tryptophan by 1.0% (w / w) 3) was produced.

(IV) Crustacean culture and high temperature tolerance test (1) Test shrimp The shrimp with an average body weight of 12.5g brought from an outdoor farm was acclimatized for one week at a water temperature of 25 ± 1 ° C. Provided.

(2) Breeding method After laying sand on the bottom of four FRP square water tanks with a capacity of 1 ton to a thickness of 20 cm, each of the above shrimps is accommodated 46 units per tank, and the water temperature is 25 ° C to 1 day. The temperature was raised by 1 ° C., and the test was started after reaching 30 ° C.

  During the test period, the temperature was raised by 1 ° C. every 1 week from a water temperature of 30 ° C. and bred at 30 to 33 ° C. for a total of 4 weeks.

(3) Feeding test
In the first group, a comparative preparation containing arginine 3.0% (w / w) and tryptophan 0.6% (w / w) was given as a control group. In the 2nd ward, the preparation 1 in which the amount of arginine was increased by 0.5% (w / w) (containing 3.5% arginine (w / w)) compared to the comparative preparation, and in the 3rd ward, 1 arginine was added compared to the comparative example. 0.02% (w / w) increase in formulation 2 (containing 4.0% (w / w) arginine), and 4 zones increased arginine by 2.0% (w / w) compared to the comparative example (arginine) Formulation 3 containing 5.0% (w / w) was given.

  From 2 days before the start of the test (water temperature 28 ° C) for 3 days (test start date, water temperature 30 ° C), the above preparations were given so that each group had a weight of 2.0% (w / w). When there was no food, it was increased by 0.5% (w / w), and when there was residual food, it was fed for 4 weeks by a method of decreasing by 0.5% (w / w).

(4) Efficacy evaluation method The number of dead tails, survival rate, residual feed, food intake, weight gain and molting status during the test period in the arginine (and tryptophan) administration group and the control group were examined and the effects were evaluated. .

(5) Test results

  Table 3 shows the test results in the arginine (and tryptophan) administration group and the control group, and FIG. 1 shows the results of changes in the survival rate after the start of the test.

  In the control group, death occurred 2 days after the start of the test when the water temperature was set to 30 ° C., and 47.8% died by the end of the test 4 weeks later. Died 6 days later than the control, and 45.7% died by the end. On the other hand, deaths were finally observed in 17 days after the start of the test in the 3 wards of the preparation 2 administration, but 10.9% died by the end of the study, and in the 4 wards of the preparation 3 administration, the deaths were seen 25 days later. Only 6.5% died by the end. There was a significant difference in the mortality and survival rate between the control group and the 2 and 3 and 4 groups (p <0.05).

  Compared with the mortality rate by water temperature, 21.7% of deaths were observed in the control group at 30 ° C, 4.3% in the second group administered with Formulation 1, and the third group treated with Formulation 2 and Formulation 3 and Although the mortality rate was 0% in the 4th group, 22.2% in the control group and 25.0% in the 2nd group administered with the preparation 1 at 31 ° C, the 3rd and 4th groups administered with the 2nd and 2nd preparations. The ward was 0%. At 32 ° C., 10.7% in the control group, 18.7% in the second group administered with the preparation 1, 2.2% in the third group administered with the preparation 2, 0% in the fourth group administered with the preparation 3, 33 ° C. In the control group, it was 4.0%, 7.4% in the second group administered with the preparation 1, 6.8% in the third group administered with the preparation 2, and 6.5% in the fourth group administered with the preparation 3.

  Based on the above facts, about half of the prawns in the control group fed with a feed formulated with amino acid growth requirements (arginine content 3.0% (w / w), tryptophan content 0.6% (w / w)) Died at 30 to 31 ° C., but formulation 1 (arginine content 3.5% (w / w), tryptophan content 0.6% (w / w)) increased in 0.5% (w / w) arginine w)) In the second administration, almost no shrimp died at 30 ° C., and about half of them died at 31-32 ° C. In addition, it survived in the third section administered with Formulation 2 (arginine content 4.0% (w / w), tryptophan content 0.6% (w / w)) increased by 1.0% (w / w) arginine. Formulation 3 in which 10.9% died in the temperature range of 32 to 33 ° C. far exceeding the limit temperature, and arginine was increased by 2.0% (w / w) and tryptophan was increased by 1.0% (w / w). No mortality was observed at 30 to 32 ° C. in the 4 sections administered with arginine content of 5.0% (w / w) and tryptophan content of 1.6% (w / w), and 6.5 at 33 ° C. % Only died.

  As described above, it has been clarified that the addition of arginine to the feed imparts resistance to the survival limit temperature in the high temperature region.

(V) Crustacean culture and low temperature tolerance test (1) Test prawns Shrimp with an average weight of 14 g brought from an outdoor farm were acclimatized for 1 week at a water temperature of 15 ± 1 ° C and then subjected to the following tests. .

(2) Breeding method After laying sand on the bottom of four FRP square water tanks with a capacity of 1 ton to a thickness of 20 cm, 50 shrimp are stored per tank, and the water temperature is from 15 ° C to 1 day. The temperature was lowered by 1 ° C., and the test was started after reaching 12 ° C.

  During the test period, the temperature was lowered by 1 ° C. every one week from the water temperature of 12 ° C., and reared at 12-7 ° C. for a total of 42 days.

(3) Feeding test In the same manner as above, a comparative preparation containing 3.0% (w / w) of arginine was given to the first section as a control section. In the second ward, preparation 1 with an arginine increase of 0.5% (w / w) compared to the comparison preparation (arginine content 3.5% (w / w)) was added, and in the third ward, arginine was added compared to the comparison preparation. Formulation 2 with 1.0% (w / w) increase (arginine content 4.0% (w / w)), and arginine 2.0% (w / w) Formulation 3 was given in which tryptophan was increased 1.0% (w / w) (arginine content 5.0% (w / w), tryptophan content 1.6% (w / w)).

  When the above preparation is given at 1% (w / w) of body weight in each group for 3 days from the day before the start of the test (water temperature 14 ° C) for 3 days (test start date, water temperature 12 ° C), after which there is no residual food Was fed by 0.5% (w / w), and when residual food was observed, the food was fed for 42 days by a method of reducing 0.5% (w / w).

(4) Efficacy evaluation method The number of dead tails, survival rate, residual food, food intake, weight gain and molting status during the test period in the arginine (and tryptophan) administration group and the control group were examined, and the effects were evaluated. .

(5) Test results

  The test results in the arginine (and tryptophan) administration group and the control group are shown in Table 4.

  No death was observed in all sections at a water temperature of 12 to 11 ° C., but at a water temperature of 10 to 9 ° C., 8% of shrimp in the control group died and There was no death. At a water temperature of 8-7 ° C., 50% of the control group died, 20% in the second administration of Formulation 1 and 4% in the third administration of Formulation 2; There was no death. The cumulative mortality rate at the end of the study was 58% in the control group, whereas the 2, 3, and 4 groups of the preparations 1 to 3 administration were 20, 4, 0%, respectively. Significant differences were observed in the mortality and survival rates of the 4 wards (p <0.05).

  In addition, there was a difference not only in mortality but also in food intake between the preparation administration group and the control group. That is, in the control group, up to a water temperature of 12 ° C., about 0.5 to 1% (w / w) of body weight per day was consumed, but not fed at 11 ° C. or less, and the total food intake was 30 Whereas it was 0.3 g, in the second section where the preparation 1 was administered, food was fed even at a water temperature of 11 ° C., and the total food intake was 75.3 g. In addition, the three sections of the preparation 2 administration were fed 0.5 to 2% (w / w) of body weight per day for 14 days set at a water temperature of 12 to 10 ° C., and the total food intake was 111. In the 4th section of Formulation 3 administration, 0.5 to 2% (w / w) was consumed per day for 15 days set at a water temperature of 12 to 10 ° C., and the total food intake was 124. .4 g.

  As described above, it has been clarified that addition of arginine to feed not only under high-temperature survival limit water temperature but also under low-temperature limit water temperature, feeds and grows even at low water temperature Was confirmed.

Example 2 Effect of Arginine-Containing Formulation on Fish (I) Production of Arginine 3.0% (w / w) -Containing Formulation

  Using the raw materials shown in Table 5a (including 3.0% (w / w) arginine derived from natural raw materials), 3.0% (w / w) arginine is contained in the same manner as in Example 1. An agent for imparting survival limit temperature resistance (Formulation 4) was produced. In this example, the comparative preparation is a fish feed containing 2.5% (w / w) arginine.

(II) Production of preparation containing arginine 4.0% (w / w)

  Using the raw materials shown in Table 5b (including arginine 3.0% (w / w) derived from natural raw materials), 4.0% (w / w) arginine is contained in the same manner as in Example 1. A survival limit temperature tolerance-imparting agent (formulation 5) was produced.

(III) Production of 5.0% (w / w) arginine-containing preparation

  Using the raw materials shown in Table 5c (including 3.0% (w / w) arginine derived from natural raw materials), 5.0% (w / w) arginine is contained in the same manner as in Example 1. An agent for imparting survival limit temperature resistance (Formulation 6) was produced.

(IV) Fish culture and high temperature tolerance test (1) Test fish Red sea bream with an average weight of 52 g brought from a seedling production site in Nagasaki Prefecture was acclimatized for a week at a water temperature of 25 ± 1 ° C and then used for the following test. did.

(2) Breeding method In the circular aquarium with a capacity of 500 liters, the above red sea bream is accommodated 30 units per ward (ie, per aquarium), and the temperature is raised from 25 ° C. to 1 ° C. per day to 29 ° C. The test started after it reached.

  During the test period, the water temperature was raised from 29 ° C. by 1 ° C. every week, and was raised to 29-33 ° C. for a total of 5 weeks.

(3) Feeding test One group was given a comparative preparation containing 2.5% (w / w) of arginine as a control group. Formulation 4 containing 3.0% (w / w) arginine in the 2nd section, Formulation 5 containing 4.0% (w / w) arginine in the 3rd section, and Arginine 5.0% in the 4th section Formulation 6 containing% (w / w) was given.

  From 2 days before the start of the test (water temperature 27 ° C.) to 3 days (the test start date, water temperature 29 ° C.), the above-mentioned preparations were given so as to be 2.0% (w / w) of body weight in each group, and then the remaining food When there was no food, the food was increased by 0.5% (w / w), and when residual food was observed, the food was fed for 5 weeks by a method of decreasing the weight by 0.5% (w / w).

(4) Effect evaluation method The number of dead tails, the survival rate, the amount of remaining food, the amount of food intake, and the weight gain during the test period in the arginine administration group and the control group were examined, and the effect of arginine was evaluated.

(5) Test results

  The test results in the arginine administration group and the control group are shown in Table 6, and the transition of the survival rate after the start of the test is shown in FIG.

  In the control group, death occurred 2 days after the start of the test when the water temperature was set to 29 ° C., 10% died by 14 days after setting the water temperature to 31 ° C., and 100% under the 32 ° C. water temperature after 23 days. Died. On the other hand, in the test section, the second section of the preparation 4 administration started to die after 17 days at 31 ° C water temperature, and 66.7% died by the end of the test, and the third section of the preparation 5 administration was at 33 ° C water temperature. Only 3.3% (1 fish) died 29 days after the end of the test, and no animals died by the end of the test in the 4th section of the preparation 6 administration, and all fish survived.

  Comparing the mortality rate by water temperature, 10% death was observed in the control group at 29-30 ° C, whereas no death was observed in any test group, and 20% in the control group at 31 ° C. %, 6.7% died in the 2nd administration of the preparation 4, but no death was observed in the 3rd administration of the preparation 5 and the 4th administration of the preparation 6. At 32 ° C., all the fish in the control group and 50% of the 2nd group administered with the preparation 4 died, whereas no death was observed in the 3rd group administered with the 5th preparation and the 4th group administered with the 6th preparation. Further, at 33 ° C., 3.3% of deaths were observed only in the 3 sections administered with the preparation 5, but no death was observed in the 4 sections of the control group and the preparation 6 administration.

  In this way, in the control group fed with the comparative preparation (arginine content 2.5% (w / w)) containing the amount of amino acids necessary for red sea bream growth, all fish died by 32 ° C. On the other hand, in the test section fed with the preparation with an increased amount of arginine, there was no death in the four sections where the preparation 6 (arginine content 5.0% (w / w)) was administered by the end of the test at 33 ° C water temperature. It was not seen.

    From the above, it has been clarified that the tolerance to the survival limit temperature in the high temperature region is imparted to fish by administering arginine exceeding the amount necessary for growth.

(V) Crustacean aquaculture / low temperature tolerance test (1) Test fish A red sea bream with an average weight of 52 g brought from a seedling production site in Nagasaki Prefecture was acclimatized for a week at a water temperature of 15 ± 1 ° C. Provided.

(2) Breeding method The red sea bream was placed in a 500 liter circular aquarium at 30 units per ward (ie per aquarium), and the temperature was lowered from 15 ° C. to 1 ° C. per day, reaching 12 ° C. Later the test started.

  During the test period, the temperature was lowered from the water temperature of 12 ° C. by 1 ° C. every week using a water temperature cooling device, and reared from 12 to 8 ° C. for 35 days.

(3) Feeding test One group was given a comparative preparation containing 2.5% (w / w) of arginine as a control group. Formulation 4 containing 3.0% (w / w) arginine in the 2nd section, Formulation 5 containing 4.0% (w / w) arginine in the 3rd section, and Arginine 5.0 in the 4th section Formulation 6 containing% (w / w) was given.

  When the above feed is given at 1% (w / w) of body weight in each section for 2 days from the start of the test (water temperature 14 ° C) for 3 days (test start date, water temperature 12 ° C), after which there is no residual food The animals were fed for 35 days by increasing 0.5% (w / w), and if residual food was observed, the amount was reduced by 0.5% (w / w).

(4) Effect evaluation method The number of dead tails, the survival rate, the amount of remaining food, the amount of food intake, and the weight gain during the test period in the arginine administration group and the control group were examined, and the effect of arginine was evaluated.

(5) Test results

  The test results in the arginine administration group and the control group are shown in Table 7.

  When the water temperature was 12 to 11 ° C., no death was observed in all groups, but at a water temperature of 10 to 9 ° C., 13.3% of red sea bream in the control group died, and 6. Although 7% died, no mortality was observed in the 3rd section administered with Formulation 5 and the 4th section administered with Formulation 6. At a water temperature of 8 ° C., 40% of red sea bream in the control group died, whereas 20% died in 2 groups of Formulation 4 administration and 6.7% died in 3 divisions of Formulation 5 administration. There were no deaths in the ward.

  In addition, there was a difference not only in the mortality rate but also in the amount of food consumed in the preparation administration group and the control group. That is, in the control group, until about 0.5 to 1.0% (w / w) of the body weight per day until the first half of the water temperature of 12 ° C. and 11 ° C., it was not fed from the latter half of 11 ° C. In contrast, the total food consumption was 161 g, whereas in the second section of the preparation 4 administration, food was fed until the latter half of 10 ° C., and the total food consumption was 266 g. In addition, in the third section of the preparation 5 administration, 0.5 to 1.5% (w / w) of body weight per day was consumed for 21 days set at a water temperature of 12 to 10 ° C., and the total food intake was 349 g. In the 4th section of the preparation 6 administration, 0.5 to 1.5% (w / w) of body weight per day was consumed for 23 days set at a water temperature of 12 to 9 ° C., and the total food intake Was 396 g.

  From the above, it has become clear that administration of a feed supplemented with increased amounts of arginine imparts resistance not only under high temperature limit water temperature but also under low temperature limit water temperature. It was observed that they could feed and grow even at water temperature.

  According to the present invention, there is provided a fish or crustacean survival temperature limit tolerance-imparting agent capable of obtaining a stable aquaculture effect, and a culture method capable of surviving fish or crustaceans even in a temperature range exceeding the survival limit water temperature range of fish and crustaceans. Provided.

  All publications, patents and patent applications cited herein are incorporated herein by reference in their entirety.

Claims (7)

  Crustacea with aquaculture water temperature of 29 ° C. or higher or 12 ° C. or lower, characterized in that arginine is added to aquatic feed so that the arginine content is 3.5% (w / w) or higher. Aquaculture methods. Crustacean prawns, Penaeus monodon, Ri Oh white leg shrimp or portunus trituberculatus, method of cultivating shellfish according to claim 1, wherein the water temperature of the aquaculture field at 29 to 33 ° C. or 7 to 12 ° C.. The method for cultivating a crustacean according to claim 1 or 2 , further comprising adding tryptophan to the aquatic feed so that the tryptophan content is 0.6% (w / w) or more.   It is characterized by adding arginine to the aquatic feed so that the arginine content is 3.5% (w / w) or more to the crustaceans where the water temperature of the farm is 29 ° C or higher or 12 ° C or lower. A method for imparting survival temperature tolerance to the crustacean.   Fish having an aquaculture water temperature of 29 ° C. or higher or 12 ° C. or lower, characterized in that arginine is added to aquatic feed so that the arginine content is 3.0% (w / w) or higher. Farming methods. Fish sea bream, flatfish, yellowtail, amberjack, tiger puffer, tuna, eel, trout or rainbow trout der is, in the water temperature is 29 to 33 ° C. or 7 to 12 ° C. farms, farming method of fish according to claim 5, wherein.   It is characterized in that arginine is added to fishery feed so that the arginine content is 3.0% (w / w) or more to fish whose water temperature in the farm is 29 ° C or higher or 12 ° C or lower. A method for imparting survival temperature tolerance to the fish.

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