CN106386590A - Method for establishing gossypol induced fish enteritis model - Google Patents
Method for establishing gossypol induced fish enteritis model Download PDFInfo
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- CN106386590A CN106386590A CN201610276358.6A CN201610276358A CN106386590A CN 106386590 A CN106386590 A CN 106386590A CN 201610276358 A CN201610276358 A CN 201610276358A CN 106386590 A CN106386590 A CN 106386590A
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- QBKSWRVVCFFDOT-UHFFFAOYSA-N gossypol Chemical compound CC(C)C1=C(O)C(O)=C(C=O)C2=C(O)C(C=3C(O)=C4C(C=O)=C(O)C(O)=C(C4=CC=3C)C(C)C)=C(C)C=C21 QBKSWRVVCFFDOT-UHFFFAOYSA-N 0.000 title claims abstract description 106
- 241000251468 Actinopterygii Species 0.000 title claims abstract description 90
- QHOPXUFELLHKAS-UHFFFAOYSA-N Thespesin Natural products CC(C)c1c(O)c(O)c2C(O)Oc3c(c(C)cc1c23)-c1c2OC(O)c3c(O)c(O)c(C(C)C)c(cc1C)c23 QHOPXUFELLHKAS-UHFFFAOYSA-N 0.000 title claims abstract description 53
- 229930000755 gossypol Natural products 0.000 title claims abstract description 53
- 229950005277 gossypol Drugs 0.000 title claims abstract description 53
- 238000000034 method Methods 0.000 title claims abstract description 20
- 208000004232 Enteritis Diseases 0.000 title abstract description 21
- 206010009887 colitis Diseases 0.000 claims description 34
- 238000002474 experimental method Methods 0.000 claims description 23
- 230000006698 induction Effects 0.000 claims description 8
- 241000252230 Ctenopharyngodon idella Species 0.000 claims description 6
- 230000037396 body weight Effects 0.000 claims description 5
- 108090000623 proteins and genes Proteins 0.000 claims description 5
- 102000004169 proteins and genes Human genes 0.000 claims description 5
- 229920002261 Corn starch Polymers 0.000 claims description 3
- 235000019733 Fish meal Nutrition 0.000 claims description 3
- 240000007594 Oryza sativa Species 0.000 claims description 3
- 235000007164 Oryza sativa Nutrition 0.000 claims description 3
- 229920002472 Starch Polymers 0.000 claims description 3
- 239000008120 corn starch Substances 0.000 claims description 3
- 229940099112 cornstarch Drugs 0.000 claims description 3
- 239000004467 fishmeal Substances 0.000 claims description 3
- 239000001963 growth medium Substances 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 235000009566 rice Nutrition 0.000 claims description 3
- 235000019698 starch Nutrition 0.000 claims description 3
- 239000008107 starch Substances 0.000 claims description 3
- 238000005453 pelletization Methods 0.000 claims description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 13
- 238000004519 manufacturing process Methods 0.000 abstract description 12
- 230000036039 immunity Effects 0.000 abstract description 4
- 210000003205 muscle Anatomy 0.000 abstract description 2
- 230000001717 pathogenic effect Effects 0.000 abstract 1
- 210000000981 epithelium Anatomy 0.000 description 14
- 235000013372 meat Nutrition 0.000 description 9
- 238000007689 inspection Methods 0.000 description 6
- 229920000742 Cotton Polymers 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 210000001842 enterocyte Anatomy 0.000 description 4
- 210000000936 intestine Anatomy 0.000 description 4
- 210000000952 spleen Anatomy 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 230000003247 decreasing effect Effects 0.000 description 3
- 230000007119 pathological manifestation Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000002477 vacuolizing effect Effects 0.000 description 3
- 108010022579 ATP dependent 26S protease Proteins 0.000 description 2
- 206010020565 Hyperaemia Diseases 0.000 description 2
- 244000046052 Phaseolus vulgaris Species 0.000 description 2
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 241000607528 Aeromonas hydrophila Species 0.000 description 1
- 241001270131 Agaricus moelleri Species 0.000 description 1
- 235000019750 Crude protein Nutrition 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 235000012343 cottonseed oil Nutrition 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 208000010824 fish disease Diseases 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 1
- 210000003750 lower gastrointestinal tract Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000029052 metamorphosis Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 230000003950 pathogenic mechanism Effects 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/20—Animals treated with compounds which are neither proteins nor nucleic acids
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/40—Fish
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
- A01K2267/035—Animal model for multifactorial diseases
- A01K2267/0368—Animal model for inflammation
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Environmental Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Hematology (AREA)
- Immunology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
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- Biotechnology (AREA)
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Abstract
The invention discloses a method for establishing a gossypol induced fish enteritis model. The method comprises the following steps: selecting normal cultured fishes as objects for establishing an enteritis model; and feeding the cultured fishes with an experimental feed in which the content of gossypol is 250mg/kg, thereby obtaining the gossypol induced fish enteritis model. The method for establishing the gossypol induced fish enteritis model, disclosed by the invention, has smallest influence on production performance, growth development of intestinal tracts, immunity and muscle quality of the fishes, but caused enteritis pathogenic manifestations are most suitable, and the model is stable and reliable, and has good reproducibility.
Description
Technical field
The present invention relates to the method for building up of animal model is and in particular to a kind of gossypol induces the method for building up of fish colitis model.
Background technology
Cotton dregs are the flour cakes that cottonseed draws after squeezing, through extract technology, most of Residual oil of the inside is separated again, the granular article of a kind of micro- red or yellow obtaining, it is the primary raw material of processing and manufacturing feed, the crude protein that it contains is up to 40%, even up to 44%, equally matched with the gross protein value of dregs of beans；The price of cotton dregs is cheap compared with dregs of beans simultaneously, wide material sources, and the cotton dregs yield of China is high, and annual production is more than 12,000,000 tons.Therefore, because the special physiological feature of fish is so that cotton dregs become the cheap protein feeds source of the important high-quality of fish feed.And the gossypol containing in cotton dregs, to cultured fishes animal pest, seriously limits it and uses in fish feed.But the current research of mechanism that gossypol produces illeffects to fish is considerably less, and which toxic action unclear its has to fish.
Enteritis is cultured fishes, such as grass carp, one of most common disease.The fatal rate of fish enteritis, generally 50%, sometimes even up to 90%, is harm cultured freshwater fish disease the most serious.This disease causes cultured fishes to be up to the catastrophic loss of several hundred million RMB every year.
According to the physilogical characteristics of cultured fishes, set up stable fish colitis model, be the research of fish enteritis pathomechanism and the medicine/of new preventing and treating/or alleviation fish enteritis or the good tool of nutrition prescription screening.And how to set up that to stablize suitable fish colitis model be current urgent problem.Under many circumstances, according to the fish colitis model being not suitable for method foundation, it is easily caused fish enteritis and becomes excessively serious and then lead to fish dead, the death rate is too high, and fish colitis model is unstable, so that fish colitis model is applied and is restricted.
Content of the invention
It is an object of the invention to provide a kind of gossypol induces the method for building up of fish colitis model so that the fish colitis model set up is stable, the enteritis pathological manifestations causing are optimal, both will not be excessively slight and will not be excessively serious.
The present invention is achieved through the following technical solutions：
A kind of gossypol induces the method for building up of fish colitis model, comprises the following steps：
Select normal cultivation fish as the object setting up colitis model；
Take the experiment feed that gossypol content is 250mg/kg, feed above-mentioned cultivation fish, obtain the fish colitis model of gossypol induction.
Present invention only requires feeding can obtain the suitable fish colitis model of fish enteritis reason performance for 60 days, and the production performance of fish colitis model, immunocompetence and the meat quality that obtain are declined slightly, without the serious growth hindering fish.
The present invention is found by research：When gossypol content is less than 250mg/kg, the production performance of fish, enteron aisle grow, the fall of immunocompetence and meat quality is less, but fish enteritis reason performance is also excessively slight；When gossypol content is higher than 250mg/kg, the production performance of fish, enteron aisle grow, immunocompetence and meat quality have significant decline, and fish enteritis reason performance is excessively serious, very serious vacuolation in gut epithelium, the enterocyte on pleat top comes off very serious, the 26S Proteasome Structure and Function of enteric epithelium is badly damaged, leads to the fish incidence of disease to raise, or even fish is dead；When gossypol content is for 250mg/kg, cause gut epithelium occur obvious cavity, the enterocyte on pleat top comes off substantially, fish enteritis manage performance be optimal, and its production performance, enteron aisle grow, immunocompetence and meat quality fall less.
A kind of gossypol induces the method for building up of fish colitis model, comprises the following steps：
Select the growth medium cultivation fish that body weight is 230-727g as the object setting up colitis model；
Gossypol is mixed in main component be fish meal, FSPC, in the feed of rice protein powder, cornstarch and alphalise starch, be configured to the experiment feed that gossypol content is 250mg/kg；
The amount that above-mentioned experiment feed is pressed 3wt% fish body weight daily feeds 60 days, obtains the fish colitis model of gossypol induction.
Wherein, prepared experiment feed, after pelletizing, is placed in 4 DEG C of temperature and saves backup.
Wherein, described cultivation fish includes but is not limited to grass carp.
The present invention compared with prior art, has such advantages as and beneficial effect：The present invention is grown on the production performance of fish, enteron aisle using the method for building up that gossypol induces fish colitis model, immunocompetence and meat quality impact are minimum, but the enteritis pathological manifestations causing are optimal, and model stability is reliable, favorable reproducibility.
Brief description
Accompanying drawing described herein is used for providing the embodiment of the present invention is further understood, and constitutes the part of the application, does not constitute the restriction to the embodiment of the present invention.In the accompanying drawings：
Fig. 1 is the microphoto of the experiment fish enteron aisle application histotomy inspection gut epithelium form of the embodiment of the present invention 1, and in the experiment feed of feeding, gossypol content is 0 mg/kg；
Fig. 2 is the microphoto of the experiment fish enteron aisle application histotomy inspection gut epithelium form of the embodiment of the present invention 2, and in the experiment feed of feeding, gossypol content is 125 mg/kg；
Fig. 3 is the microphoto of the experiment fish enteron aisle application histotomy inspection gut epithelium form of the embodiment of the present invention 3, and in the experiment feed of feeding, gossypol content is 250 mg/kg；
Fig. 4 is the microphoto of the experiment fish enteron aisle application histotomy inspection gut epithelium form of the embodiment of the present invention 4, and in the experiment feed of feeding, gossypol content is 375 mg/kg；
Fig. 5 is the microphoto of the experiment fish enteron aisle application histotomy inspection gut epithelium form of the embodiment of the present invention 5, and in the experiment feed of feeding, gossypol content is 775 mg/kg；
Fig. 6 is the microphoto of the experiment fish enteron aisle application histotomy inspection gut epithelium form of the embodiment of the present invention 6, and in the experiment feed of feeding, gossypol content is 1175 mg/kg.
Specific embodiment
For making the object, technical solutions and advantages of the present invention become more apparent, with reference to embodiment and accompanying drawing, the present invention is described in further detail, and the exemplary embodiment of the present invention and its explanation are only used for explaining the present invention, not as a limitation of the invention.
Embodiment l-6：A kind of method for building up of the fish colitis model of gossypol induction, comprises the following steps：
1st, according to a conventional method using growth medium grass carp 540 tail of 230-727g as the object setting up colitis model, be randomly divided into 6 treatment groups, every group 3 is repeated, each repeat 30 tail fishes.
2nd, the experiment feed containing gossypol for the preparation
Gossypol is uniformly mixed in main component be fish meal, FSPC, in the feed of rice protein powder, cornstarch and alphalise starch, be configured to respectively gossypol content be 0,125,250,375,775 and 1175mg/kg experiment feed, pelletize, be placed in 4 DEG C of refrigerators and save backup；
3rd, the foundation of model
The grass carp that step 1 is selected is raised in 1 cubic metre of net cage, feeds the experiment feed 60 days prepared by above-mentioned steps 2 daily as the amount of 3wt% fish body weight respectively, obtains the fish colitis model of gossypol induction.
The fish colitis model product test that above-described embodiment l-6 obtains is as follows：
The gossypol induction fish colitis model of 4.1 pairs of embodiment l-6 acquisitions measures production performance, enteron aisle grows and meat quality, and result is recorded in table 1 below.
Table 1
Note：In upper table 1, FE is feed efficiency, can reflect the utilization ratio of feed；SGR is specific rate, can reflect the speed of growth of fish；ISI and ILI is respectively intestines body ratio and index of the intestines length, can reflect growing of enteron aisle；Shearing force and pH can reflect the quality of muscle；Subscript a, b, c, d, e are to characterize the significance of difference between each group of data using the labelled notation of Multiple range test, the labelled notation of this Multiple range test be those skilled in the art commonly use mark the method whether between each data with significant difference.
As can be seen that after gossypol content is more than 250mg/kg, the production performance of fish is remarkably decreased, and enteron aisle grows and is seriously obstructed, and meat quality is decreased obviously from upper table 1.When gossypol content is less than or equal to 250mg/kg, the production performance of fish, enteron aisle grow and meat quality is all declined slightly, but impact is less.
The gossypol induction fish colitis model of 4.2 pairs of embodiment l-6 acquisitions carries out attacking poison by the way of lumbar injection Aeromonas hydrophila, to detect fish enteron aisle, spleen and head-kidney immunity and germ resistivity, shown in its result table specific as follows 2.
Table 2
Note：In upper table 2, subscript a, b, c, d, e are the significances of difference to be characterized using the labelled notation of Multiple range test between each group of data.
Can be seen that from upper table 2, after gossypol content is more than 250mg/kg, the enteron aisle of fish, spleen and head-kidney immunity and germ resistivity are remarkably decreased, and anterior intestine, middle intestines and hindgut red and swollen hyperemia incidence significantly raises, and spleen and the head-kidney hyperemia enlargement incidence of disease also dramatically increase.And when gossypol content is less than or equal to 250mg/kg, the enteron aisle of fish, spleen and head-kidney immunity and germ resistivity are declined slightly.
The gossypol of 4.3 pairs of embodiment l-6 acquisitions induces fish colitis model enteron aisle application histotomy to check the metamorphosis of gut epithelium, and its result is as shown in figures 1 to 6.It can be seen that, as shown in figure 1, when gossypol content is 0 mg/kg, gut epithelium morphosis is normal；As shown in Fig. 2 when gossypol content is more than 0 mg/kg and is less than 250mg/kg, slight vacuolation in intestinal epithelial cell, slightly coming off occurs in pleat both sides epithelial cell, and part pleat top is changed into flat-top, and fish enteritis reason performance is excessively slight；As shown in figure 3, when gossypol content is for 250mg/kg, gut epithelium obvious cavity and increases, and the enterocyte on pleat top comes off substantially, fish enteritis reason performance is optimal；As Figure 4-Figure 6, after gossypol content is more than 250mg/kg, very serious vacuolation in gut epithelium, the enterocyte on pleat top comes off very serious, the 26S Proteasome Structure and Function of enteric epithelium is badly damaged, fish enteritis reason performance is excessively serious, leads to the incidence of disease significantly to raise, or even it is dead fish.
To sum up, the present invention is grown on the production performance of fish, enteron aisle using the method for building up that the gossypol of 250mg/kg induces fish colitis model, immunocompetence and meat quality impact are minimum, but the enteritis pathological manifestations causing are optimal, model stability is reliable, favorable reproducibility.And method of the present invention can be widely used in various cultivation fishes, the including but not limited to grass carp cited by embodiment.
Above-described specific embodiment; the purpose of the present invention, technical scheme and beneficial effect are further described; be should be understood that; the foregoing is only the specific embodiment of the present invention; the protection domain being not intended to limit the present invention; all any modification, equivalent substitution and improvement within the spirit and principles in the present invention, done etc., should be included within the scope of the present invention.
Claims (4)
1. a kind of gossypol induces the method for building up of fish colitis model it is characterised in that comprising the following steps：
Select normal cultivation fish as the object setting up colitis model；
Take the experiment feed that gossypol content is 250mg/kg, feed above-mentioned cultivation fish, obtain the fish colitis model of gossypol induction.
2. a kind of gossypol induces the method for building up of fish colitis model it is characterised in that comprising the following steps：
Select the growth medium cultivation fish that body weight is 230-727g as the object setting up colitis model；
Gossypol is mixed in main component be fish meal, FSPC, in the feed of rice protein powder, cornstarch and alphalise starch, be configured to the experiment feed that gossypol content is 250mg/kg；
The amount that above-mentioned experiment feed is pressed 3wt% fish body weight daily feeds 60 days, obtains the fish colitis model of gossypol induction.
3. gossypol according to claim 2 induces the method for building up of fish colitis model it is characterised in that prepared experiment feed, after pelletizing, is placed in 4 DEG C of temperature and saves backup.
4. gossypol according to claim 2 induces the method for building up of fish colitis model it is characterised in that described cultivation fish includes but is not limited to grass carp.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610276358.6A CN106386590B (en) | 2016-04-29 | 2016-04-29 | A kind of method for building up of gossypol induction fish colitis model |
| FR1753666A FR3050614B1 (en) | 2016-04-29 | 2017-04-27 | METHOD FOR ESTABLISHING A MODEL OF FISH ENTERITIS INDUCED BY GOSSYPOL. |
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| CN201610276358.6A CN106386590B (en) | 2016-04-29 | 2016-04-29 | A kind of method for building up of gossypol induction fish colitis model |
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Cited By (2)
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| CN110447787A (en) * | 2018-05-07 | 2019-11-15 | 中国科学院水生生物研究所 | Galanthamine is preventing and treating the application in fish enteritis caused by bean-dregs feed |
| CN113040070A (en) * | 2020-12-15 | 2021-06-29 | 苏州大学 | Method for establishing grass carp intestinal flora imbalance model |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116369249B (en) * | 2023-01-13 | 2024-05-17 | 四川轻化工大学 | Construction method of zebra fish enteritis model |
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| CN110447787A (en) * | 2018-05-07 | 2019-11-15 | 中国科学院水生生物研究所 | Galanthamine is preventing and treating the application in fish enteritis caused by bean-dregs feed |
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| Publication number | Publication date |
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| FR3050614A1 (en) | 2017-11-03 |
| CN106386590B (en) | 2019-03-19 |
| FR3050614B1 (en) | 2023-01-13 |
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