JPH1143441A - Preventive and therapeutic agent for disease of fish and shellfish - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a therapeutic agent useful for preventing and treating infection diseases of fishes and shellfishes caused by bacteria and as an antimicrobial for foods by formulating a chitosan derivative obtained by reacting chitosan with saccharides bearing a reductive terminal. SOLUTION: The preventive and therapeutic agent for diseases of fishes and shellfishes is obtained by formulating a chitosan derivative prepared by reacting chitosan with saccharides, preferably aldoses and ketoses (e.g. glucose, galactose, etc.), bearing a reductive terminal. The objective therapeutic agent possesses proliferation repressive effect against Vibrio bacteria, Edwardsiella bacteria, Pasteurella bacteria, Aeromonas bacteria and Streptococci. As the concentration of the chitosan derivative in a medicinal solution, a culture pond and a tub, the concentration of 0.1-100 mg/ml is preferably suitable. When the mixed feed is administrated, the chitosan derivative is added to the feed for cultivation of fishes and shellfishes preferably in an amount of 1-20 wt.% to feed to fishes and shellfishes. The feed may be administrated at least once a day at a daily dose of 0.5-5 wt.% of the fish body weight.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel agent for preventing and treating fish and shellfish diseases containing a chitosan derivative obtained by reacting chitosan with a saccharide having a reducing terminal, and an application thereof.

[0002] Its field of application is that it can be used as a prophylactic or therapeutic agent for fish and shellfish diseases, such as prevention and treatment of infectious diseases caused by various bacteria, and antibacterial agents for foods (Vibrio bacteria, Edwajella bacteria, Pasteurella bacteria, Aeromonas). Bacteria, streptococci)
Also available as

[0003]

2. Description of the Related Art In recent years, aquaculture has become more and more active due to the 200 nautical mile problem and the shortage of fishery resources due to overfishing. As a result, high-density breeding of fish and shellfish has been promoted, and especially hamachi and eel. In high-density aquaculture fisheries conducted in such areas, the outbreak frequency of fish and shellfish infectious diseases is high,
If left unattended, it can cause near-total annihilation, and even if the annihilation is avoided, the surviving seafood will suffer a lot of water and damage, and the product value will be significantly reduced. I have. Even if it is commercialized without any problem, there is a danger that humans may be infected with Vibrio parahaemolyticus food poisoning due to raw sashimi of fish and shellfish, which is harmful.

[0004] As a method for preventing and treating infectious diseases of fish and shellfish, for example, administration of kasugamycin for vibrio disease and tuberculosis of Hamachi (Japanese Patent Application Laid-Open No. 61-275219) and administration of bile acid for streptococcal infection ( JP-A-62-5
9214), and administration of thianphenicols for vibrio disease and gypsum disease (Japanese Patent Application Laid-Open No. 48-56812).

[0005] However, most of the above-mentioned disease prevention methods involve large doses of antibiotics and nutrients.
It has many problems, such as the effects of fish and shellfish on the living body (toxicity and side effects), the safety of humans and the emergence of resistant bacteria, and the economics of antibiotics and nutrients themselves, making them simple and effective. The development of a new alternative was desired.

Therefore, various drugs have been used for preventing and treating infectious diseases of fish and shellfish inside and outside the body. Antibiotics and synthetic antibacterial preservatives which occupy most of them have been used.
Certainly, the effect is strong, but there are problems in terms of safety, and although it is effective, it requires side-effects such as side effects. Therefore, effective substances with few side effects have been demanded.

[0007]

Accordingly, the present inventors have developed various antibacterial and antiseptic effects against infectious diseases and diseases caused by bacteria and which are safe for use in fish and shellfish. We have been studying natural extracts and studying new substances and their effects.

[0008]

Means for Solving the Problems In view of the above circumstances, the present inventors have developed various types of chitosan derivatives obtained by reacting chitosan or a low molecular weight chitosan with a saccharide having a reducing terminal. A novel antibacterial and antiseptic effect against various bacteria (vibrio, edwajella, pasteurella, eromonas, streptococcus) and safe against seafood The present inventors have found that application to a therapeutic agent is extremely effective, and completed the present invention. Hereinafter, the process leading to the present invention will be described.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The chitosan of the present invention, the saccharide having a reducing end, the production method, and the like are described below, but are not limited thereto.

The chitosan of the present invention is usually obtained by deacetylating chitin obtained from crabs and shrimp crustaceans by alkali treatment, enzymatic treatment, etc., and the degree of deacetylation is not particularly limited. Those having a degree of 50% or more are preferred.

The low molecular weight chitosan can be obtained, for example, by treating chitosan with concentrated hydrochloric acid or chlorine gas, sodium perborate, or hydrogen peroxide, or by treating chitin with an acid or alkali, and simultaneously with deacetylation. It may be obtained by a method of degrading glycosides or a method of hydrolyzing with enzymes such as chitinase.

Examples of the saccharide having a reducing end include aldoses and ketoses such as glucose, galactose, and the like.
Monosaccharides include mannose, arabinose, xylose, erythrose and glyceraldehyde, heptulose, hexulose and pentulose, and also amino sugars such as glucosamine, N-acetylglucosamine and galactosamine, deoxy sugars and branched sugars. Further, disaccharides to polysaccharides containing these monosaccharides as components, such as maltose, isomaltose, maltotriose, lactose or various oligosaccharides, pullulan, inulin, amylose,
Examples include amylopectin, dextran, dextrin, natural polysaccharides of starch, and the like, and their decomposed products, isomerized products, and derivatives (such as induction of COOH grouping).
One or more species can be used for the reaction.

The chitosan derivative of the present invention refers to a saccharide having a 2-position amino group of glucoseamine, which is a constituent sugar of chitosan, and a reducing terminal, sodium cyanoborohydride (NaCNBH ₃ ) or an amine borane-based reducing agent such as Ammonia borane (NH ₃ BH ₃ ), monomethylamine borane “CH
₃ (NH ₂ BH ₃ ) ”or the like to form an amide bond between a saccharide having a carboxyl group terminal opened and / or a saccharide having a carboxyl group terminal released in a molecule. Things.

For the formation of the amide bond, SOCl ₂ is allowed to act on the terminal of the free carboxyl group of the above-mentioned saccharide, or esterified with p-nitrophenol, or acid chloride with phosphorus pentoxide or the like to activate the carboxyl group. The chitosan derivative can also be obtained by reacting with the amino group at the 2-position of glucose amine, which is a constituent sugar of chitosan. Use a coupling agent if necessary.
Alternatively, a reducing agent may be used in combination with a system containing chitosan and a saccharide having a reducing terminal to release a carboxyl group and simultaneously form an amide bond.

As a method for activating a free carboxyl group of a saccharide having a reducing end, for example, D-galactopyranosyl-gluconic acid of a disaccharide may be used to activate the carboxyl group in the form of p-nitrophenyl ester. Activate,
After separation of the activating compound, it is reacted with the free amino groups of chitosan. This reaction is carried out in a solvent such as dimethylformamide (DMF), tetrahydrofuran (THF), or dimethylsulfoxide (DMSO) at room temperature or under cooling.
The reaction time is completed in several hours to several days.

Next, as the coupling agent, reactivity,
In consideration of selectivity, a carbodiimide-based coupling agent such as N, N-dicyclohexylcarbodiimide (DCC), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and the like can be used. ,
One or more of these coupling agents can be used in the reaction.

Further, as for the reducing agent, reactivity and selectivity are considered.
Considering sodium cyanoborohydride (NaCNBH_Three) Or
Amine borane-based reducing agents such as ammonia borane (NH_ThreeB
H_Three), Monomethylamine borane "CH_ThreeNH_TwoBH_Three”
Luamine borane `` (CH<sub>Three</sub>)<sub>Two</sub>NHBH<sub>Three</sub> ) ", Trimethylamine
Run `` (CH_Three)_ThreeNBH_Three) ", Pyridine borane (C_FiveH_FiveNBH_Three)
And one or two of these reducing agents.
The above can also be used for the reaction.

Further, the hydroxyl group and amino group still released of the chitosan compound having a sugar side chain introduced thereinto were subjected to, for example, oxyethylene, oxypropylene addition, carboxymethylation, phosphorylation, sulfated, dihydropropylated, and the like. Derivatives may be used.

The chitosan derivative obtained as described above may be further purified by washing with a solvent, forming a precipitate, or purifying by gel filtration, ion exchange column chromatography, activated carbon exchange column chromatography, cellulose dialysis membrane, or the like. Can also.

The rate of derivatization of the saccharide having a reducing terminal with respect to chitosan of the present invention can be appropriately selected according to the purpose and application, but generally, it is preferably 10 to 100%.

Further, the chitosan derivative of the present invention is applied to prevent and treat fish and shellfish diseases by drying, concentrating or diluting them arbitrarily according to the form and dosage form of the fish and shellfish.

The method for treating or preventing bacterial diseases of fish and shellfish using the chitosan derivative of the present invention as a prophylactic / therapeutic agent for fish and shellfish diseases is not specifically defined.
The fish or shellfish may be bathed mainly in a drug solution containing the chitosan derivative of the present invention, or the chitosan derivative may be dissolved and cultured in a culture pond or tank. Is also good.

Further, the concentration of the chitosan derivative in the drug solution or the culture pond or tank is usually from 0.10.1 to 1000 mg / ml,
Preferably, 0.1 to 100 mg / ml is appropriate. The time of bathing the fish and shellfish in the chitosan derivative liquid drug solution is not particularly limited, but usually 1 to 3 hours can be exemplified.
In the case of feed mixing administration, the chitosan derivative is added to the fish and shellfish culture feed at 0.01 to 50% by weight, preferably 1 to 50% by weight.
20% by weight is added to feed the fish and shellfish. Addition to the feed may be performed at the time of preparing the feed in the case of a compound feed, or may be performed at the time of feeding when using a commercially available fish feed. The feeding amount can be appropriately selected depending on the fish species, fish age, body weight, and the like. Usually, 0.5 to 5.0% of the fish body weight may be administered once a day or more.

The prophylactic / therapeutic agent for fish and shellfish diseases containing the chitosan derivative of the present invention may be used as it is, but is usually mixed with various diluents, for example, pellets, powders, particles, granules, tablets, capsules. , Solutions, emulsions, dispersions and the like. or,
The diluent may be solid or liquid, and suitable diluents include, for example, water, ethanol, sucrose, corn oil, starch, talc, pentonite, fish meal, sodium carbonate, sodium bicarbonate and the like. Further, nutrients, feeds, other antibacterial agents, insecticides, metabolic regulators and the like may be arbitrarily selected and used in combination.

As other additives to the prophylactic / therapeutic agent for diseases of fish and shellfish, the following additives derived from plant or animal raw materials can be used. can do. In addition, various drug effects of each raw material known so far were expected, and by combining them, the intended effect was enhanced or multifunctional effects were expected. It can also be a product.

Specific plants (crude drugs) as raw materials include:
For example, almond (Hen peach), eye (Aiba), blue oak
(Seifu Fuji), Aotsuji Lafzi (Kibo), Acacia, Akasuri Fruit, Red Grape, Red Megawiwa (Red Bud), Akane
(Kanekane Akane), Ayajiou Jiou (Jihuang), Agi (Awei),
Akinire (Royu skin), Akebi (Kidori), Asa (John Asako), Maruba Asagao or Asagao (Toshiko), Ashitaba (Asuka),
Azuki (red azuki), acerola, ase yak (Asenyaku), anise, Abemaki fruit, avocado, ama, amacha (sweet tea),
Amachaworu, Amadokoro (Jade bamboo), Amigasa lily (Shellfish), Altea, Arnica, Aloe (lux), Aloe vera, Angelica, Anzu Honganzu (Anjin), Ansokkou (Azuka), Ikakozolina (Biliary head), Itadori
(Tiger cane root), strawberry, fig (fructose) or its leaves, strawberry (winter aoiko), ichiyakuso, ginkgo (ginko leaf, ginkgo),
Itohimehagi (Enshi), carob, rice seed or seed coat,
Inondo seeds, Ibuki-kusou-so, nettle, ylang-ylang, fennel (Fenka), ukiyagara (sanryou), gull-kasukagra fruit, Hime-uikyo, turmeric (Utsukin), Usubasaishin-keirinsaishin (spicy), Usveniaoi,
Pterodactyla mushroom, Nepenthes (summer hay), Udo or Shishudo (Qang, Independent, Tang Independent), Plum (Ume) or its pulp, Vladigashi, Uwaurushi, Unshuri mandarin (Chen skin), Estragon, Ezocogi (Ezo fivega), Enoki ,
Ebisugusa (Keiko), Elderberry fruit, Elemi, Enju (Gossip, Gossip rice), Ogi-kibana-ohgi (Koki),
Ouren (yellow), giant crow squirrel (karonin), giant skull (self-defense), psyllium (vegetative child, vegetative grass), toucan hazel (Haruko), obanokera okera (white jitsu), obayashishabushi fruit, o hawthorn hawthorn ), Barley (barley), oguruma (turning over), okra fruit, panax ginseng and tochibanningi (carrot), hypericum persicae, gogomebato perforated flowers, perforated periwinkle (brother cut grass), odorikosou (interrupted), onamimimi (small earlings) (Tianhe), Onyuri Sasayuri Hakatayuri (Yuri), Ononis, Halibut (Hihayu), Ominaeshi (Loose), Dutch strawberry, Dutch mustard (Cresson), Dutch jelly, Dutch honey, olive, oregano, orange fruit or Pericarp, lycaenium (ripe yellow), cacao seeds, oysters or their leaves (persimmon), Fujikagi), Cassia, paper mulberry fruit (Kozomi), Zedoary (莪朮), oak
(Kazuki, Kazaha), Cascarilla, Kanixa (Kinosato), Kanokosou (Kisakune), Birch or Weeping birch (White birch), Pumpkin, Kapokinoki seeds, Kahoku Sansho (shopper), Gama (Kaba yellow), Chamomile roma Chamomile, kamiyatsude (weeds), crow squirrel (flower meadow), crow squirrel (midsummer), oak wheat, guarana seeds, karaya, karin (gourd), kawayanagi, kawataketake, kawarayomogi (inchinko), kansui
(Sweet), liquorice (licorice), candelilla, cantou, canna, raspberry, kiwi fruit, kikarasuri (urorone), kikyo (bellflower, bellflower root), chrysanthemum (chrysanthemum flower), catalpa (azumi),
Rumex (Yotei), Pheasant (Kikomi), Kizuta, Kidachi Aloe, Kidachiha, Kinoki, Yellowfin (Huangbashi), Gymnema sylvestre, Cabbage, Cabbage immature, Cucumber,
Kiraja saponaria, Kiraya, kumquat fruit, betel, kinmizukiki (sensuru-kusa), guava fruit, guayule,
Quercus Infectoria (Gallery), Kukui Nuts, Kukui (Gugi, Gyugia, Gyugoli, Leaf Skin), Kusagi Kazura (Tenmen Fuyu), Kuzu (Kakuzu), Kusunoki, Gooseberry Fruit, Gardenia, Kunugi Saw), Kumazasa, Kumatsuzura (horse whipweed), Clara (Bittersweet), Cranberry fruit,
Chestnut or its astringent skin, Curculigo latifolia fruit, grapefruit fruit, buckthorn, Black stalk
(Rescue), cloves (cloves, cloves), kayga (crop, crocodile spikes), celosia (cow flowers, rooster), bay laurel (laurel), gentiana, gennoshoko (old crocodile), koushinbara (moon season flowers) , Kosui mint (Melissa), Kozo fruit, Kocha (tea), Kouhone (Raw bone), Kouhon (Straw and Kara straw), Koulyan, Koendro fruit, Koganebana (Yellow Gon), Cowberry (Koshibachi), Coconut fruit, Goshyu
(Goshuyu), goshoichigo (covered bonko), pepper (pepper), copaiba balsam, coffee beans, kobushi tamshiba magnolia (spicy), burdock (burdock, burdock), kobotdur, sesame (sesame), sesame crab (gensin) ), Wheat (wheat), rice, rice bran (red bran, white bran), rice oil, cola acuminata seed, cola vera seed, fenugreek fruit, colombo, conzulango, kelp, konjac, comfrey (finnari grass), saxima Button ヅ (weird sen), cherry leaves, flowers, fruits, bark
(Cherry bark), cherry, pomegranate, sasa, lily
Sazanka, Sajiomodaka (Sawahka), Sweet potato, Sugarcane, Sugar beet, Sanebu jujube (Acid jujube), Saffron (Bamboo safflower, West safflower), Pomelo fruit, Sabonsou, Salmonberry fruit, Sarashinashoma (Masame), Salvia (Sage), Sawagikyo (Yamakona), Hawthorn (Yamazako), Panax ginseng (Seven carrots), Sanshuyu (Yasugoyu), Sansho (Sansho), Sandscon (Yamazone), Shea (Kareite), Cyanoki fruit, Shiitake ( Shiitake mushrooms), Shion (Shien), Dikitaris, Shikunshi (Kimiko), Shiso, Aodiso, Chirimenjiso, Catamendiso (Shiso leaves, Shiso), Rosewood, Shinano,
Chinese Forsythia (Forsythia), Zinnia (Chrysanthemum), Spiraea, Potato, Peony (Shakuyaku), Shajin (Shasan), Janohige (Bakumonyu), Juzudama, Palm Fruit,
Ginger (ginger), ginger (iris, iris root), shizuku fruit, birch oak seeds, white walnut butterflies, white blossoms, white blossoms, white blossoms, white blossoms, white cinnamon syllabus, cinnamon, watermelon, watermelon (Gold and silver flowers, Shinobi winter), sorrel (acid model), swimberry fruit, strawberry fruit, horsetail (Thorn), stevia, plum fruit, pine cone, cone larch,
Vines, walnuts, hawthorn,
Dandelion, horse chestnut (malonie), pear fruit, prunus japonicus, prunus japonicus, prunus edulis, prunus edulis, millefoil, balsam pears, pygmy root, pygmye pylon, ), Mallow, hirohasenega, senega, senega, seri, celery, senkyu
(River Kyu), Sendan, Senburi (Drug), Senna fruit or leaves, Senninsou (Large Tate), Soka (Grass), Buckwheat seeds,
Somenimo, Daio (daioh), Radish, Soybean, Daidai
(Orange, kikomi), takatodai (daigeki), takarababi (guin), dark sweet cherry fruit, hollyhock, hollyhock thyme (thyme, momoka), hollyhock (tachibana skin), hollyhock willow, tamarind seeds, onion, tamshiba ),
Tara tree or its root bark, tanjin (dansin), dandelion (gakko) or white dandelion and moko dandelion, dammar, cherry fruit, tigaya or its root (chinese), chikutsujin carrot (bamboo ginseng), chicory, ginseng (gomiko) , Korean ginseng (Rhuanghuang), Korean ginseng (Wuyi), Korean ginseng (Ginseng), Korean ginseng (Aiba leaf), Chinese ginseng (Irei), Japanese radish, Camellia,
Butterflies, clovers (lacquer primroses), twigs (ducklings), vines (red azuki), vines (red bean), vines, vines (vegetables), vines (four-leaf ginseng), vines, deco, teuchigurumi, duberry fruit, tengusa, tendaiyaku
(Crowd medicine), Pepper (winter melon), Pepper (bansho), Touki
(Return), calendula (marigold), corn or corn hair (southern hair), horned rattle (female Sadako), linden (dragonfly), dokudami (jujutsu), konkon (gem), tochibanjin (bamboo ginseng) ), Eucommia (Tonaka, Tonaka leaf), Tomato, Traganth, Triacanthus seed, Tormentilla, Dronoki, Troroaoi, Niseria berry fruit, Nagaimo yam (Yakuyaku), Nagiikada (Butcher bloom), Naginata kouji, Nazuna, Rapeseed, Natsumikan , Jujube (large jujube), naniwaiba (gold cherry), narco lily (yellow), peanut (peanut), nanten (minamiminami), nigaki (maki), sagebrush (bite), nikuzuku, nikkei (cinnamon), leek ( Nirako), elderberry (bone-bonded tree)
Fruit, flower or foliage, garlic (garlic), nurde (five times), leeks, thistle (daiji), wild rose (fruit), sawwood, nodake (maego), nobara, peach, palm, pineapple fruit, hibiscus (Bussouge) , Furinbussouge, roselle), hakata lily (lily), hakamauraboshi (bone crushing supplement), haxen (ringworm skin), huckleberry fruit,
Chickweed (Horiko), Hazel (Haruko), Hashidokoro (Roth Root), Basil, Lotus (Russian meat, Renko), Parsley (Netherland jelly), Hadamugi, Batata, Hachiku Madake (bamboo boiling), Patchouli, Mint ( Light-loading leaves), Adlay (Yokuinin), Hanasuge (Chimbo), Banana, Hanamint, Vanillabins, Papaya, Dipterocarpus (mouse koji), Hub grass, Paprika, Hamago and Mitsuba Hamago (Twilight), Hamasuge (Katsukishi) , Hamabishi (Shikoriko), Hamanas (Maikai flower),
Hamabofu (beach storm), Hamamelis, Rose (rose), Parietaria, Harunire (Yellow, Yuhakuh, Yuba), Breadfruit,
Hiogi (sun dried), lizard carrot (party), pecan nuts, gangamana (stone and vines), cypress (extended life grass), cypress (hishomi), pistachio, beet, hitotsuba (stone reed), hinotakokochi ( Beef knee), hinoki, hiba, castor, sunflower, bell pepper, hemima, incense, himematsutake, pimenta fruit, juniper, hiyu fruit, belowo oyster, hirohaokinagusa (bakuohoku), loquat (loquat, loquat leaf),
Betel nut (large clothing skin, betel nut), sugar beetle (southern rattan), butterflies, coltsfoot (winter flowers, winter leaves), fujibakama (orchid grass), wisteria bean (tomato), grape fruit or pericarp / leaf, beech, fumulus mushroom (Cordyceps), Brazilian liquorice, black currant fruit, blackberry, plum fruit, flucellaria, blueberry (Acacia catechu), prunes, blond psyllium, bundou (mung bean), luffa, safflower (safflower), belladonna, berry fruit, boysen Berry fruit, boufu (windbreak), spinach, physalis (Torone), honoki (kopoku, sumkopaku), bokeh
(Gourd melon), Hosobananaokera (blue ajutsu), Hosobabarengiku, Bodaiju (bodaiju), button (peony, peony bark), hop, jojoba, whiteberry fruit, honshimeji, meizurutennanshaw (Tennan star), ephedra (mao), macadamia nut , Makuri (sea grass), Magwa (mulberry bark, mulberry leaf), mushroom
(Bamboo-boiled), matabi (mokuten tate), matsukasa, matsuba, matsuhodo
(Bukuryo), mayola (Hanaminka), malvanozadine (bitter), mulberry fruit, quince, malonie, mango, mangosteen, manshugulumi, mandarin fruit, mannentake (reishi), citrus fruit (kikomi), and mishimasaiko (saiko) ), Mizokokushi (half lotus), misohagi (senkakuna), mitsugashiwa, honeywort, honeysuckle maggot (vines), midorihakka, mimosa, mioga, miracle fruit fruit, myrrh, mirobaran, mukuge, mukunoki, mukuroji , Murasaki (purple root), Murasaki corn, Japanese swordfish (Essentiale grass), Mebogi, Melaleuca, Melissa, Meliloth, Melon fruit, Japanese mugwort, Moso bamboo, Mokkou (wood incense), Japanese maple, Peach leaf
(Peach leaves), fruits, seeds (peach kernels), bean sprouts, morello cherry fruits, moroheiya (burlap), yakawamurasakiimo, yakuchi
(Masutoshi), cornflower (cornflower), cornflower mint, yashabushi (arrow wheel), yachiyanagi, yatsude (octagonal lantern), mistletoe (yanagi parasite), willow leaf, yabugarashi, yabukouji (purple gold cow), yamagobo (land) ), Mountain alder (Yamaharu), bayberry (Yumebai), yamamogi, eucalyptus, yukinoshita (tiger ear grass), yucca flebifolia, yuzu fruit, lily, yorigusa, mugwort (autumn leaf), lime fruit, rye, lacanca fruit, raspberry Leaf / fruit, lavender, longan (longan meat), ryocha (green tea), apple fruit, gentian, lubas suavisimus (sweeten), lettuce, red currant fruit, lemon fruit, lemongrass, forsythia, astragalus, wax palm , Loganberry fruit, Logwood, Rosemary (Mannenrow), Rosehip (Nobara), Wakame, Wa Bi, and the like burnet (Chinire).

Examples of the seaweeds include seaweeds [green algae: Chlorella vulgaris, Chlorella pyrenoidosa, Chlorella ellipsoidia, Usubaaonori, Suziahonori, Hiraaonori, Bouuaonori, Hosoedaaonori], and seaweeds [Brown algae: Macombum, Lyconib, Hosoeabu] Honey kelp, wakame, hirome, blue seaweed, macrocystis pilifera, macrocystis integrifolia, neocitys luetokeana, hijiki, hibamata], seaweed [red algae: red alga: jijirimen, maxa (tengusa), hirakusa, onyxa, obaxa, kataobaxa ,
Yatabegusa, Yuikiri, Shimatengusa, Tosakanori, Togekirinsai, Akumasakirinsai, Kirinsai, Byaksinkirinsai, Tsunomata, Obatsunomata, Tochaka (Yahatsunomatata), Ezotunomata, Togetsunomata, Hirakotonogi, Kotoji, Kotoji , Heius Banori] and the like.

Materials derived from animal raw materials include cockscomb extract, bovine / human placenta extract, swine / cattle stomach, duodenum or intestinal extract or its decomposition product, and swine / cattle spleen extract. Product or its degradation product, extract of pig / bovine brain tissue, water-soluble collagen, collagen derivative such as acylated collagen, collagen hydrolyzate, elastin, elastin hydrolyzate, water-soluble elastin derivative, keratin and its decomposition product Or a derivative thereof, a silk protein or a degradation product thereof or a derivative thereof, a pig / bovine blood cell protein degradation product (globin peptide), a pig / bovine hemoglobin degradation product (hemin, hematin,
Heme, protohem, heme iron), milk, casein or its decomposition products or their derivatives, skim milk powder or its decomposition products or their derivatives, lactoferrin or its decomposition products, chicken egg components, fish meat decomposition products and the like.

(1) Microbial culture metabolite Yeast metabolite, yeast extract, rice ferment extract, rice bran extract, euglena extract, lactic acid fermentation product of skim milk powder, trehalose or a derivative thereof.

(2) Flavors Natural animal flavors such as musk, civet, castorium, ambergris, anise essential oil, angelica essential oil, ylang-ylang essential oil, iris essential oil, fennel essential oil, orange essential oil, cananga essential oil, caraway essential oil, cardamom essential oil, Guayakwood essential oil, cumin essential oil, black letter essential oil, cinnamon essential oil, cinnamon essential oil, geranium essential oil, copaiba balsam essential oil, coriandel essential oil, perilla essential oil, cedarwood essential oil, citronella essential oil, jasmine essential oil, gingergrass essential oil, cedar essential oil, spearmint essential oil, western Mentha essential oil, Great Fengxiang essential oil, Tuberose essential oil, T-shaped essential oil, Orange flower essential oil, Winter green essential oil, Truval sam essential oil, Batturi essential oil, Rose essential oil, Palmarosa essential oil, Cypress essential oil, Hiba essential oil,
Sandalwood essential oil, Petit glen essential oil, Bay essential oil, Vetiva essential oil, Bergamot essential oil, Peruvian balsam essential oil, Boad rose essential oil, Camphor essential oil, Mandarin essential oil, Eucalyptus essential oil, Lime essential oil, Lavender essential oil, Linaloe essential oil, Lemongrass essential oil, Lemon essential oil, Rosemary Vegetable flavors such as essential oils and Japanese peppermint essential oils, and other synthetic flavors.

(3) Dyes / colorants Red cabbage dye, red rice dye, madder dye, anato dye, squid dye, turmeric dye, ende dye, krill dye, persimmon dye, caramel, gold, silver, gardenia dye, corn dye , Onion pigment, tamarind pigment, spirulina pigment, buckwheat whole plant pigment, cherry pigment, laver pigment, hibiscus pigment, grape juice pigment, marigold pigment, purple potato pigment, purple yam pigment, lac pigment, rutin, etc.

(4) Sweeteners Sugar, sweet tea, fructose, arabinose, galactose, xylose, mannose, maltose, honey, glucose, miraculin, monelin, licorice extract and the like.

(5) Various vitamins Vitamin A group: retinol, retinal (vitamin
A₁), Dehydroretinal (vitamin A_Two), Carochi
N, lycopene (provitamin A), vitamin B group: thia
Min hydrochloride, thiamine sulfate (vitamin B₁), Lvov
Rabin (vitamin B_Two ), Pyridoxine (vitamin
B₆), Cyanocobalamin (vitamin B₁₂), Folic acid,
Nicotinic acids, pantothenic acids, biotins, choline,
Inositols, vitamin C group: ascorbic acid or its
Derivatives, vitamin D group: Ergocalciferol (bi
Tamin D_Two), Cholecalciferol (vitamin D_Three),
Dihydrotaxerol, Vitamin E group: Tocophero
Or its derivatives, ubiquinones, vitamin K group:
Tonadione (vitamin K₁), Menaquinone (vitamin
K_Two), Menadione (vitamin K_Three), Menadiol (bi
Tamin K_Four), Other essential fatty acids (vitamin F), mosquitoes
Lunitine, ferulic acid, γ-oryzanol, orot
Acids, vitamin Ps (rutin, eriocitrin, hesperi
Gin), vitamin U, etc.

(6) Various amino acids Valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, glycine, alanine, asparagine, glutamine, serine, cysteine, cystine, tyrosine, proline, hydroxyproline, aspartic acid, glutamic acid , Hydroxylysine, arginine, ornithine, histidine and the like, and their derivatives such as sulfates, phosphates, nitrates, citrates, and pyrrolidone carboxylic acids.

(7) Nutrition enhancer Shell calcined calcium, cyanocorabamine, yeast, wheat germ, egg yolk powder, hemicellulose, heme iron and the like.

(8) Dairy products Milk, cheese, fresh cream, butter, margarine, milk powder, whey, condensed milk and the like.

In addition, humectants, hormones, pH adjusters, cooling agents, stabilizers, emulsifiers, animal and vegetable proteins and their degradation products, animal and vegetable polysaccharides and their degradation products, animal and vegetable sugars Proteins and their degradation products, blood flow enhancers, anti-inflammatory and anti-allergic agents, cell activators, keratolytic agents, wound healing agents,
Foaming agents, thickeners, oral agents, deodorants / deodorants, bitters, seasonings, enzymes, etc.
Various additive and synergistic effects can be expected.

The dosage form of the prophylactic / therapeutic agent for fish and shellfish diseases containing the chitosan derivative of the present invention is arbitrary,
Capsule, pill, tablet, powder, granule, solid,
The composition can be used in various forms such as liquid, gel, foam, emulsion, cream, and sheet.

Further, the use of the prophylactic / therapeutic agent for fish and shellfish diseases containing the chitosan derivative of the present invention is described in, for example, sea bream (Thailand), yellowtail (burri), hamachi, horse mackerel, horse mackerel, bonito (bonito), tuna (tuna) ), Saury, mackerel, mackerel, trout, trout, salmon, flatfish, flounder, river pig (pufferfish), carp (koi), sweetfish (ayu), sunfish (carriage fish), whale (whale) ), Eel, catfish, catfish,
Freshwater or seawater fish such as shrimp (shrimp), and more, eel (sazae), abalone (abalone), oyster (oyster), red mussel (red mussel), scallop (scallop), akoya mussel (pearl oyster), clam ( Antibacterial agents (vibrio bacteria, etc.) for shellfish such as clams and clams, and other general foods and beverages
(Edowella, Pasteurella, Aeromonas, Streptococcus).

[0040]

EXAMPLES The present invention will be described below with reference to production examples and test examples, but the present invention is not limited thereto.

(Production Example 1) 5.0 g of D-galactopyranosyl-gluconic acid was dissolved in 30 ml of TEMED buffer (50 mM tetramethylenediamine, PH4.7), and 5.0 g of carbodiimide hydrochloride (EDC) was added. Stir for a minute. Next, 10 g of a chitosan (manufactured by Yaizu Suisan Kagaku Kogyo KK, 33% deacetylation degree) was dissolved in 10 g of TEMED buffer solution 12 in this EDC solution.
Add 0 ml and stir for 3 days. During this time, 3.3 g of EDC is added in several portions. Thereafter, low molecular components were removed by dialysis, washed and dried to obtain about 12.5 g of a chitosan derivative of the present invention (chitosan-lactose derivative: about 31.5% of lactose induction, confirmed by ¹³ C-NMR).

(Production Example 2) 150 ml of water was added to 10 g of chitosan (manufactured by Yaizu Suisan Chemical Co., Ltd., 85% deacetylation degree), stirred, dispersed, and adjusted to pH 11 with an aqueous sodium hydroxide solution.
After adjusting to, the mixture is heated to 70 ° C. Next, 5.25 ml of hydrogen peroxide (about 35% concentration) was added and reacted for 3 hours. After the reaction was completed, the reaction solution was removed, washed with water and dried to obtain low molecular weight chitosan (deacetylation degree 89%). About 8.0 g was obtained. Next, 1.0 g of this low-molecular-weight chitosan is placed in a 300 ml flask, and 50 ml of a mixed solution of methanol / 1% acetic acid aqueous solution = 1/1 is added and dissolved. 5.0 g of maltose is added and dissolved, and then cyanoborohydride is further added. 2.5 g of sodium was added and reacted for 5 hours. After completion of the reaction, the mixture was washed with 300 ml of a mixed solution of acetone / water = 5/1 and 300 ml of ether and dried, and the chitosan derivative of the present invention (chitosan-maltose derivative: maltose induction ratio about 42.5%) was reduced to about 1.7%.
g obtained.

(Test 1) Vibrio, Edwajella,
Growth inhibition test against Pasteurella, Aeromonas, and Streptococcus [Test bacterial species] 1. Vibro anguillarum vibrio bacteria 2. Etodishella tarda (Edwardsiella tarda) 3. Pasteurella piscicida Pasteurella. 4. Aeromonas samonicida, Aeromonas samonicida Streptococcus equinus Streptococcus [Media used] Brain Heart Infusion Broth (BH)
I) "Eiken" [Preparation of sample bacterial solution] Culture the bacteria used for the test in the above medium
(37 ° C., 18 hours), and diluted with the same medium so that the number of bacteria was 1.0 × 10 ⁵ . [Preparation of sample] Chitosan derivative of the present invention (Production Example 1)
3181 g was dissolved in purified water, made up to 50 ml, and filtered through a 0.45 μm membrane filter to obtain a sample solution. [Test Method] Purified water was used as a diluent in advance, and a multiple dilution series of the sample solution was prepared (0.5 ml). 0.4 ml of the above medium and 0.1 ml of the test bacterial solution were added thereto, and the mixture was cultured at 37 ° C for 18 hours.
The MIC (minimum inhibitory concentration) was determined. The results are shown in Table 1.

[0044]

[Table 1]

[Test Results] The chitosan derivative of the present invention was tested against MIC against Vibrio bacteria, Aeromonas bacteria and Streptococci.
(Minimum growth inhibitory concentration) was 0.16 w / v% or less, and the growth inhibitory effect could be confirmed with a small amount. In addition, the growth inhibitory effect was able to be confirmed with a small amount against E. swallowa and Pasteurella. Similarly, the chitosan derivative of the present invention (Production Example 2) is also effective against Vibrio bacteria, Edwajella bacteria, Pasteurella bacteria, Aeromonas bacteria, and Streptococci.
A growth inhibitory effect almost equivalent to that of Production Example 1 was confirmed.

(Test 2) Safety test (1) Acute toxicity test Similarly, the chitosan derivative of the present invention obtained in Production Examples 1 and 2 was purified water so as to have a dry solid concentration of about 10 ^W / _V %. Orally administered to ddy mice (male and female, 5 mice / group, 5 weeks old) in a dose of 2,000 mg / kg prepared for 4 hours before the test. Observed. As a result, no abnormalities were observed in all the mice for 14 days, and no abnormalities were observed in the dissection results. Therefore, LD ₅₀
It was determined to be 2,000 mg / kg or more.

(Prescription Examples) Production of Preventive / Treatment Agents for Fish and Shellfish Diseases The following prescription examples are shown below according to the above evaluation results. Each prescription example may be one produced by a conventional method in the production of each product. Only shown. Further, the present invention is not limited to these.

(Formulation Example 1) Soluble powder for medicine bath The chitosan derivative powder (1-10%) of Production Example 1 and lactose (99-90%) are mixed and uniformized.

(Formulation example 2) Powder mixed with feed 5% of the chitosan derivative powder of Production Example 1, 90% of soybean powder and 5% of aloe extract are mixed to prepare a feed.

(Formulation example 3) Powder mixed with feed 3% of the chitosan derivative powder of Production Example 2, 5% of shrimp, 5% of seaweed, 12% of lecithin and 70% of gluten are mixed to prepare a feed.

(Test 3) Use effect test (Test to prevent the onset of Vibrio disease in Thailand) Two 100-liter water tanks were prepared, and 40 liters of breeding water at 24 ° C. were added to each of them. The chitosan derivative of the invention was dissolved, and the other aquarium was usually filled with breeding water. Vibrio anguillarum at a concentration of 10 ⁸ cfu / ml in 80 ties with an average body weight of 0.4 kg
After allowing the bacteria to bath in suspension breeding water for 2 hours, add
The animals were housed in 0 tails and reared for 10 days, and the onset of Vibrio disease (damage to the body surface and the tip of the mouth, tumor formation in the subcutaneous muscle and visceral organs, etc.) was observed 10 days later. In addition, the disease-causing status was observed in the same manner using a water-raised fish containing no chitosan derivative as a control.

[Test Results] As a result, the number of diseased fish after breeding for 10 days was only one in the chitosan derivative-containing water-raising fish of the present invention, whereas the number of chitosan derivative-free water was 10%. The number of the reared fish was 36, and the disease-inhibiting effect of rearing in water containing the chitosan derivative was confirmed.

(Test 4) Usage effect test (test for inhibiting disease of Streptococcus infestation of Hamachi) Two 100-liter water tanks were prepared, and 40 liters of breeding water at 24 ° C. was added to each of them, and 1 g of water was added to one of the water tanks. Of the present invention was dissolved, and the other aquarium was filled with ordinary breeding water. 80 tails of yellowtail average weight 0.5kg, 10 ⁸ cfu / ml
Concentration of Streptococcus etinus (Streptococ
cus equinus) After germ bathing in suspension breeding water for 2 hours, 40 hamachi were housed in both aquariums and bred for 10 days, and 10 days later the onset of streptococcal sensitization (redness, erosion, tumor formation on the body) ,
Observation of the situation). In addition, the disease development was observed in the same manner using a water-raised fish containing no chitosan derivative as a control.

[Test Results] As a result, the number of diseased fish after breeding for 10 days was only two in the chitosan derivative-containing water-raising fish of the present invention, whereas the chitosan derivative-free water The number of fish reared was 39, and the disease-inhibiting effect of rearing with chitosan derivative-containing water was confirmed.

[0055]

The chitosan derivative obtained by reacting the chitosan of the present invention with a saccharide having a reducing end can be used for various bacteria (Vibrio, Edwajella, Pasteurella,
E. aureus, streptococci), and is safe for humans and animals. Therefore, when used as a prophylactic or therapeutic agent for fish and shellfish diseases, disease contamination of various fish and shellfish can be prevented.

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Takehito Naito 153, 380 Iwai, Gifu City, Gifu Prefecture (72) Inventor Yoshiharu Matahira 1510-2 Higashicho, Shimada City, Shizuoka Prefecture (72) Inventor Hirofumi Yura Kanagawa Prefecture 5-9-1, Shonandai, Fujisawa-shi Town Court Room 601

Claims (2)

[Claims] 1. A preventive / therapeutic agent for a fish and shellfish disease comprising a chitosan derivative obtained by reacting chitosan with a saccharide having a reducing terminal. 2. The prophylactic / therapeutic agent for a seafood disease according to claim 1, which has an inhibitory effect on the growth of Vibrio, Edwajella, Pasteurella, Aeromonas, and Streptococcus.