CN110402854B - High-survival-rate cultivation method for fries of cupfish with round mouth - Google Patents
High-survival-rate cultivation method for fries of cupfish with round mouth Download PDFInfo
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- CN110402854B CN110402854B CN201910734311.3A CN201910734311A CN110402854B CN 110402854 B CN110402854 B CN 110402854B CN 201910734311 A CN201910734311 A CN 201910734311A CN 110402854 B CN110402854 B CN 110402854B
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- 210000000214 Mouth Anatomy 0.000 title claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 98
- 239000000463 material Substances 0.000 claims abstract description 62
- 241000607720 Serratia Species 0.000 claims abstract description 58
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicium dioxide Chemical class O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 48
- 239000002207 metabolite Substances 0.000 claims abstract description 34
- 229910052782 aluminium Inorganic materials 0.000 claims abstract description 21
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminum Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims abstract description 21
- BPQQTUXANYXVAA-UHFFFAOYSA-N silicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 claims abstract description 21
- OZECDDHOAMNMQI-UHFFFAOYSA-H cerium(3+);trisulfate Chemical compound [Ce+3].[Ce+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O OZECDDHOAMNMQI-UHFFFAOYSA-H 0.000 claims abstract description 20
- 229910052708 sodium Inorganic materials 0.000 claims abstract description 20
- 239000011734 sodium Substances 0.000 claims abstract description 20
- 235000020681 well water Nutrition 0.000 claims abstract description 18
- 239000002349 well water Substances 0.000 claims abstract description 18
- 230000003213 activating Effects 0.000 claims abstract description 16
- 238000011068 load Methods 0.000 claims abstract description 9
- CZMRCDWAGMRECN-UGDNZRGBSA-N D-sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 50
- 238000000855 fermentation Methods 0.000 claims description 39
- 230000004151 fermentation Effects 0.000 claims description 39
- CZMRCDWAGMRECN-GDQSFJPYSA-N Sucrose Natural products O([C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1)[C@@]1(CO)[C@H](O)[C@@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-GDQSFJPYSA-N 0.000 claims description 38
- 239000005720 sucrose Substances 0.000 claims description 38
- 239000001963 growth media Substances 0.000 claims description 33
- 241000132906 Tubificidae Species 0.000 claims description 32
- 210000000538 Tail Anatomy 0.000 claims description 31
- 238000002360 preparation method Methods 0.000 claims description 22
- 238000003756 stirring Methods 0.000 claims description 21
- 239000002609 media Substances 0.000 claims description 19
- 241001482311 Trionychidae Species 0.000 claims description 16
- 239000012153 distilled water Substances 0.000 claims description 16
- 238000001035 drying Methods 0.000 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 15
- GVGUFUZHNYFZLC-UHFFFAOYSA-N dodecyl benzenesulfonate;sodium Chemical compound [Na].CCCCCCCCCCCCOS(=O)(=O)C1=CC=CC=C1 GVGUFUZHNYFZLC-UHFFFAOYSA-N 0.000 claims description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 14
- 229940080264 sodium dodecylbenzenesulfonate Drugs 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 13
- 238000002156 mixing Methods 0.000 claims description 13
- 230000001376 precipitating Effects 0.000 claims description 13
- 229940041514 Candida albicans extract Drugs 0.000 claims description 12
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 12
- 239000012138 yeast extract Substances 0.000 claims description 12
- KEAYESYHFKHZAL-UHFFFAOYSA-N sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 claims description 11
- 230000000249 desinfective Effects 0.000 claims description 9
- 238000006213 oxygenation reaction Methods 0.000 claims description 9
- CPKVUHPKYQGHMW-UHFFFAOYSA-N 1-ethenylpyrrolidin-2-one;molecular iodine Chemical compound II.C=CN1CCCC1=O CPKVUHPKYQGHMW-UHFFFAOYSA-N 0.000 claims description 8
- 229920000153 Povidone-iodine Polymers 0.000 claims description 8
- 238000000498 ball milling Methods 0.000 claims description 8
- 230000003203 everyday Effects 0.000 claims description 8
- 239000011152 fibreglass Substances 0.000 claims description 8
- 230000002503 metabolic Effects 0.000 claims description 8
- 229960001621 povidone-iodine Drugs 0.000 claims description 8
- 239000000047 product Substances 0.000 claims description 8
- 238000002791 soaking Methods 0.000 claims description 8
- 239000005695 Ammonium acetate Substances 0.000 claims description 7
- 229920001353 Dextrin Polymers 0.000 claims description 7
- 239000004375 Dextrin Substances 0.000 claims description 7
- 239000001888 Peptone Substances 0.000 claims description 7
- 108010080698 Peptones Proteins 0.000 claims description 7
- 229920002472 Starch Polymers 0.000 claims description 7
- USFZMSVCRYTOJT-UHFFFAOYSA-N ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 claims description 7
- 229940043376 ammonium acetate Drugs 0.000 claims description 7
- 235000019257 ammonium acetate Nutrition 0.000 claims description 7
- 239000001110 calcium chloride Substances 0.000 claims description 7
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 7
- 239000005018 casein Substances 0.000 claims description 7
- 235000021240 caseins Nutrition 0.000 claims description 7
- 235000019425 dextrin Nutrition 0.000 claims description 7
- 235000019441 ethanol Nutrition 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 7
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 239000012299 nitrogen atmosphere Substances 0.000 claims description 7
- 235000019319 peptone Nutrition 0.000 claims description 7
- 239000011780 sodium chloride Substances 0.000 claims description 7
- 235000019698 starch Nutrition 0.000 claims description 7
- 239000008107 starch Substances 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 7
- 238000005406 washing Methods 0.000 claims description 7
- 230000001954 sterilising Effects 0.000 claims description 4
- BAUYGSIQEAFULO-UHFFFAOYSA-L Iron(II) sulfate Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L magnesium sulphate Substances [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 2
- 239000002131 composite material Substances 0.000 claims 1
- 235000019341 magnesium sulphate Nutrition 0.000 claims 1
- 239000011148 porous material Substances 0.000 claims 1
- 241000251468 Actinopterygii Species 0.000 abstract description 21
- 230000004083 survival Effects 0.000 abstract description 15
- RYGMFSIKBFXOCR-UHFFFAOYSA-N copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 abstract description 10
- 229910052802 copper Inorganic materials 0.000 abstract description 10
- 239000010949 copper Substances 0.000 abstract description 10
- 241001124569 Lycaenidae Species 0.000 abstract description 9
- 235000014987 copper Nutrition 0.000 abstract description 9
- 201000010099 disease Diseases 0.000 abstract description 3
- 230000001717 pathogenic Effects 0.000 abstract description 3
- 230000000875 corresponding Effects 0.000 abstract description 2
- 244000052769 pathogens Species 0.000 abstract description 2
- 229960004793 Sucrose Drugs 0.000 description 48
- VZJVWSHVAAUDKD-UHFFFAOYSA-N Potassium permanganate Chemical compound [K+].[O-][Mn](=O)(=O)=O VZJVWSHVAAUDKD-UHFFFAOYSA-N 0.000 description 14
- 238000011081 inoculation Methods 0.000 description 12
- 238000002372 labelling Methods 0.000 description 12
- 241000607715 Serratia marcescens Species 0.000 description 11
- 229940098362 Serratia marcescens Drugs 0.000 description 11
- 230000000052 comparative effect Effects 0.000 description 10
- 241000595940 Notostraca Species 0.000 description 8
- 230000004913 activation Effects 0.000 description 7
- 238000005273 aeration Methods 0.000 description 7
- BFNBIHQBYMNNAN-UHFFFAOYSA-N Ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 6
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 6
- 235000011130 ammonium sulphate Nutrition 0.000 description 6
- 238000005119 centrifugation Methods 0.000 description 6
- 230000004907 flux Effects 0.000 description 6
- 101710009228 grisea Proteins 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 238000001556 precipitation Methods 0.000 description 6
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Inorganic materials [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 229910052564 epsomite Inorganic materials 0.000 description 5
- 230000001488 breeding Effects 0.000 description 4
- 230000036039 immunity Effects 0.000 description 3
- 210000003128 Head Anatomy 0.000 description 2
- 241000219000 Populus Species 0.000 description 2
- 238000005189 flocculation Methods 0.000 description 2
- 230000016615 flocculation Effects 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- DRXYRSRECMWYAV-UHFFFAOYSA-N nitrooxymercury Chemical compound [Hg+].[O-][N+]([O-])=O DRXYRSRECMWYAV-UHFFFAOYSA-N 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 235000015763 Artemisia ludoviciana Nutrition 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000272201 Columbiformes Species 0.000 description 1
- 241000033272 Coreius guichenoti Species 0.000 description 1
- 206010011409 Cross infection Diseases 0.000 description 1
- 241000252231 Cyprinus Species 0.000 description 1
- 241000608847 Gnaphalium Species 0.000 description 1
- 235000005058 Madhuca longifolia Nutrition 0.000 description 1
- 240000004212 Madhuca longifolia Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000000844 anti-bacterial Effects 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 241001233037 catfish Species 0.000 description 1
- 230000005591 charge neutralization Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 230000003311 flocculating Effects 0.000 description 1
- 230000002068 genetic Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000001264 neutralization Effects 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000010408 sweeping Methods 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 238000003911 water pollution Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/22—Animal feeding-stuffs from material of animal origin from fish
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/28—Treatment of water, waste water, or sewage by sorption
- C02F1/281—Treatment of water, waste water, or sewage by sorption using inorganic sorbents
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/28—Treatment of water, waste water, or sewage by sorption
- C02F1/286—Treatment of water, waste water, or sewage by sorption using natural organic sorbents or derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/50—Treatment of water, waste water, or sewage by addition or application of a germicide or by oligodynamic treatment
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/52—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities
- C02F1/5236—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities using inorganic agents
- C02F1/5245—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities using inorganic agents using basic salts, e.g. of aluminium and iron
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Abstract
The invention provides a high-survival-rate cultivation method for round-mouth copper fish fries, which adopts well water as a water source and has few pathogens for cultivating the round-mouth copper fish. Aiming at the physiological requirements of the round-mouth coppers fry in different stages, different feeds are provided in different stages, palatable corresponding baits are adopted, the growth speed of the fry is effectively improved, the fry is strong, the physique is good, the disease resistance is good, and the survival rate of the fry is high. The water bottom material is obtained by loading polymeric aluminum ferric silicate, cerium sulfate, sodium humate and a serratia grignard metabolite (obtained by activating, inoculating and fermenting serratia grignard ACCC 01695) on modified diatomite. Improve the water environment, greatly reduce the morbidity and improve the survival rate.
Description
Technical Field
The invention relates to the technical field of aquaculture, in particular to a high-survival-rate cultivation method for fries of round-mouth coppers.
Background
Coreius guichenoti (Sauvage et Dabry) belongs to Cyprinus, subfamily gobiocyparidae and genus Cultix, and has the local name: square head, watertight son, round mouth, fat tuo, square head watertight son, sand-shelled crucian (Minjiang), Annao cat fish (Yunan Domei), pigeon fish, huzi fish (Jinshajiang midstream), and Mahua fish. It is commonly found in water systems such as the upper reaches of Yangtze river, the middle and lower reaches of Jialing river, Tuo river, Jinsha river and the lower reaches of Wujiang river (Dingrui, 1995; Liule Hei, et al, 1990), and is a special fish and an important economic fish in the upper reaches of Yangtze river. The round-mouth coppers are tender, fat and rich in fat (Diehua, 1995), and are important economical fishes at the upstream of Yangtze river and important fishing objects in the river fishery industry.
However, due to the influence of factors such as damage to the spawning site of the round-mouth copper fishes caused by over-fishing and power station construction and water pollution caused by rapid industrial development, wild resources of the round-mouth copper fishes are rapidly reduced (Yangzhi, 2009), and therefore species protection work for the round-mouth copper fishes needs to be carried out urgently. The artificial propagation and proliferation releasing of fish is one of the effective measures for species protection and population increase at present. The perfect collection and domestication, artificial breeding and large-scale fry breeding of wild parent fishes are the prerequisite conditions for ensuring the smooth operation of artificial propagation and releasing of fishes.
The current research reports on round-mouth coppers mainly relate to biology (Yuzhitang et al, 1984; Zhengxumin, 1988; Lin, 1990; Zhengxumin and Wuqing, 1998; Zhanghua et al, 1999; Kudzuvine, et al, 2001; Zhang Fang et al, 2005), physiological biochemistry (Wang Yoghui et al, 2005; Lirong et al, 2008; Liujun et al, 2008; Wu et al, 2008), molecular genetics (Liao Xiaolin, 2006; Xushuixing, 2007; Yushiping et al, 2008; hole flame, 2010), resource quantity (Yugong Liang et al, 2002; poplar, 2009; poplar et al, 2011), ecological habits (Liu le and et al, 1990; Cao Wen Xuan, 2008) and the artificial breeding of round-mouth coppers is still in a primary research stage, and the conventional fry breeding method is highly pathogenic and the survival rate.
Disclosure of Invention
The invention aims to provide a high-survival-rate cultivation method for fries of round-mouth coppers, which is simple and convenient to operate, reduces the morbidity of the fries and improves the survival rate of the fries.
In order to achieve the purpose, the invention is realized by the following scheme:
a high-survival-rate cultivation method for fries of cupfish with round mouth comprises the following steps:
(1) selecting a culture container;
(2) water quality management: paving a layer of water bottom material at the water bottom, taking non-polluted well water as a culture water source for micro-flowing water culture, and aerating all day long by adopting an oxygenation pump;
(3) stocking the fry, and feeding with different baits at different stages;
the water bottom material is obtained by loading polymeric aluminum ferric silicate, cerium sulfate, sodium humate and a serratia grignard metabolite on modified diatomite, and the serratia grignard metabolite is obtained by activating, inoculating and fermenting serratia grignard ACCC 01695.
Preferably, the thickness of the underwater material is 5-8 cm.
Preferably, the preparation method of the metabolic product of the serratia griffithii comprises the following steps: activating the serratia griffithii ACCC01695, inoculating the activated serratia griffithii ACCC01695 into a seed culture medium, and culturing at 28-31 ℃ for 12-15 hours to obtain a strain seed solution; inoculating strain seed liquid into a fermentation culture medium, carrying out shaking culture at 27-31 ℃ and 180-200 rpm/min, centrifuging, collecting fermentation supernatant, and precipitating to obtain a serratia griffithii metabolite; wherein, the formula of the seed culture medium is as follows: 8-12 g of casein peptone, 6-9 g of soluble starch, 5-8 g of dextrin, 3-5 g of sodium chloride, 2-4 g of ammonium acetate and FeSO4 0.2～0.3g，CaCl23-4 g of distilled water, 900-1100 mL of distilled water, 6.8-7.2 of pH, and sterilizing for 20-30 minutes at 121 ℃; the formula of the fermentation medium is as follows: 4-6 g of yeast extract,13c-labeled sucrose 0.2-0.3 g, MgSO4·7H2O5～8g，NaNO35-8 g, 8-12 g of KCl and 900-1100 mL of distilled water, wherein the pH value is 6.8-7.2, and the mixture is sterilized at 121 ℃ for 20-30 minutes;13c-tag end at sucrose exhaustionAnd (5) culturing.
Further preferably, the activation treatment is carried out by using an LB culture medium, and the specific method comprises the following steps: taking out the serratia griffithii ACCC01695 from the slant surface of the test tube for preserving strains, transferring the strains to an LB culture medium, wherein the inoculation amount is 2-3% by volume, and carrying out inverted culture in a constant temperature box at 28 ℃ for 24-72 hours.
Further preferably, the fermentation culture is aeration culture, and the air flux is 10-12L/min.
Further preferably, the inoculation amount of the strain seed liquid in the fermentation medium is 6-8% by volume.
It is further preferred that 1/2 formula amount is added in preparation of fermentation medium13C labeling sucrose, monitoring during shaking culture13C the dosage of the cane sugar marked by the mark,13with additional remainder added in batches when sucrose marked C is exhausted13C labeling sucrose until all13C, marking that all the cane sugar is exhausted, and finishing the culture;13the adding times of the C-marked sucrose are 3-5 times, and the adding amount is the same every time.
Further preferably, the centrifugation conditions are: centrifuging at 8000-12000 rpm/min for 15-20 min; the precipitation method comprises the following steps: and precipitating and collecting by using ammonium sulfate with the mass concentration of 50-80%.
Preferably, the preparation method of the modified diatomite comprises the following steps: adding 1 part of sodium dodecyl benzene sulfonate into 5-8 times of water by weight, stirring for dissolving, then stirring and heating to 70-80 ℃, adding 2-3 parts of activated diatomite, stirring for 3-4 hours under nitrogen atmosphere, filtering, washing with water, and drying to obtain the sodium dodecyl benzene sulfonate.
Preferably, the activated diatomite is obtained by adding diatomite into water with the weight 5-6 times that of the diatomite, performing microwave treatment for 3-5 minutes at 800-1000W, centrifuging, and drying at 120-130 ℃ for 2-3 hours.
Further preferably, the drying process conditions are as follows: drying at 60-70 ℃ for 12-18 hours.
Preferably, the preparation method of the underwater material comprises the following steps: pouring 1 part of polymeric aluminum ferric silicate, 0.1-0.2 part of cerium sulfate, 3-4 parts of sodium humate and 0.3-0.4 part of serratia grignard metabolite into 15-20 parts of absolute ethyl alcohol by weight, carrying out first ultrasonic oscillation and full ball milling, then adding 8-10 parts of modified diatomite, carrying out second ultrasonic oscillation, carrying out microwave treatment, carrying out third ultrasonic oscillation, and naturally volatilizing the ethyl alcohol to obtain the underwater material.
More preferably, the treatment time of the first ultrasonic oscillation, the second ultrasonic oscillation and the third ultrasonic oscillation is 30 to 40 minutes, 10 to 15 minutes and 30 to 40 minutes, respectively.
Further preferably, the microwave treatment process conditions are as follows: microwave treatment is carried out for 3-5 minutes at 600-800W.
Preferably, in the step (1), a circular glass fiber reinforced plastic cultivation tank with the diameter of 2m and the depth of 0.8-1 m is selected as a cultivation container.
Preferably, in the step (2), the culture container is disinfected in advance and then injected into non-polluted well water, the water depth is 0.6-0.8 m, and further preferably, the culture container is disinfected by 10ppm potassium permanganate 1-2 days in advance.
Preferably, the specific method of step (3) is: firstly, erecting a net cage in a culture container, selecting a round-mouth copperfish water flower seedling with a film outlet of 4-5 days, and putting the round-mouth copperfish water flower seedling into the net cage for culture, wherein the density is 2000-3000 tails/m3Feeding the fries for 4 times every day at the temperature of 18-22 ℃, feeding 2-3 g/ten thousand of fairy bugs in 1-10 days, and feeding 4-5 g/ten thousand of fairy bugs in 11-20 days; after 10 days, thinning the fry and transferring the fry from the net cage to a culture container for direct culture, wherein the density is 1000-1500 tails/m3Feeding after 20 days to about 2cm, disinfecting by adopting chopped tubificidae, and after 30 days, thinning the fry with the density of 500-800 tails/m3After 60 days, the height reaches about 4cm, and then, adopting soft-shelled turtle materials and tubificidae according to the mass ratio of 1: 2-4 mixing and making dough for feeding.
Further preferably, the aperture of the net cage is smaller than 1.0mm, and the size of the net cage is 1.4m by 0.6 m.
Further preferably, the feeding is carried out for 4 times per day, and the feeding time is 8:00, 12:00, 18:00 and 24: 00.
Further preferably, the size of the chopped tubificidae is less than 1mm, and the tubificidae is disinfected by soaking in 5ppm povidone iodine for 30 minutes; the protein content of the soft-shelled turtle material is more than 45 percent.
The invention has the beneficial effects that:
according to the invention, a layer of water bottom material is laid on the water bottom, pollution-free well water is used as a culture water source for micro-flowing water culture, an oxygenation pump is adopted for aeration all day long, the fry is put in culture, different baits are adopted for feeding in different stages, the incidence of the fry is greatly reduced through the culture method, and the survival rate of the fry is improved. The method comprises the following specific steps:
1. well water is used as a water source, and the number of pathogens is small when round-mouth coppers are cultured. The cudweed is easy to be infected with ichthyophthiriasis when cultured by river and lake water sources, no obvious effective treatment method except for mercurous nitrate (forbidden) exists at present, and the death rate is high in conventional culture.
2. Aiming at the physiological requirements of the round-mouth coppers fry in different stages, different feeds are provided in different stages, palatable corresponding baits are adopted, the growth speed of the fry is effectively improved, the fry is strong, the physique is good, the disease resistance is good, and the survival rate of the fry is high.
3. The water bottom material is obtained by loading polymeric aluminum ferric silicate, cerium sulfate, sodium humate and a serratia grignard metabolite (obtained by activating, inoculating and fermenting serratia grignard ACCC 01695) on modified diatomite. The polymeric aluminum ferric silicate has high ion degree and has double functions of flocculation and disinfection; after ball milling treatment, cerium sulfate has strong electric neutralization and net catching and sweeping effects, so that the flocculation effect is enhanced; the sodium humate has strong adsorption capacity and colloidal property, and can promote the activation and absorption of feed ingredients; the metabolite of the serratia griffithii has the bactericidal effect, can improve the immunity of the round-mouth copper fish, greatly reduces the morbidity and improves the survival rate. The polymeric aluminum ferric silicate, the cerium sulfate and the sodium humate synergistically purify the water body, improve the water body environment, greatly reduce the morbidity and improve the survival rate. The modified diatomite is loaded, has a porous structure and a large specific surface area, plays a similar slow release role on adsorbing and flocculating pollutants in a water body and purifying the water body on one hand, and plays a similar slow release role on the loaded polymeric aluminum ferric silicate, cerium sulfate, sodium humate, Serratia grisea metabolites and the like on the other hand, and the polymeric aluminum ferric silicate, the cerium sulfate, the sodium humate, the Serratia grisea metabolites and the like are slowly released for a long time, so that the effects are exerted for a long time, and the effect of reducing the morbidity is played.
4. And a small-size culture container is adopted, so that the cross infection of diseases is effectively reduced.
5. The laying thickness of the underwater material is adapted to the water depth, and the water body cannot be well purified due to too small thickness ratio, so that the morbidity is increased, and the survival rate of the fry is influenced; the fish fry with too large thickness occupies the living space of the round copper fish fry, and the survival rate of the fish fry is lowered.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention relates to a serratia grignard bacterium ACCC01695 which is purchased from China agricultural microorganism strain preservation management center.
Example 1
A high-survival-rate cultivation method for fries of cupfish with round mouth comprises the following steps:
(1) selecting a culture container;
(2) water quality management: paving a layer of water bottom material at the water bottom, taking non-polluted well water as a culture water source for micro-flowing water culture, and aerating all day long by adopting an oxygenation pump;
(3) stocking the fry, and feeding with different baits at different stages;
the water bottom material is obtained by loading polymeric aluminum ferric silicate, cerium sulfate, sodium humate and a serratia grignard metabolite on modified diatomite, wherein the serratia grignard metabolite is obtained by activating, inoculating and fermenting serratia grignard ACCC 01695. The thickness of the bed material was 5 cm.
The preparation method of the metabolic product of the serratia griffithii comprises the following steps: activating the serratia griffithii ACCC01695, inoculating into a seed culture medium, and culturing at 28 deg.C for 12 hr to obtain strain seed solution; inoculating strain seed liquid in fermentation culture medium at 27 deg.C and 180 deg.CPerforming shaking culture at rpm/min, centrifuging, collecting fermentation supernatant, and precipitating to obtain a metabolic product of Serratia griffonii; wherein, the formula of the seed culture medium is as follows: 8g of casein peptone, 6g of soluble starch, 5g of dextrin, 3g of sodium chloride, 2g of ammonium acetate and FeSO40.2g，CaCl23g, 900mL of distilled water, pH 6.8, and sterilizing at 121 ℃ for 20 minutes; the formula of the fermentation medium is as follows: 4g of yeast extract is prepared by mixing 4g of yeast extract,13c-labeled sucrose 0.2g, MgSO4·7H2O 5g，NaNO35g, KCl 8g and 900mL of distilled water, wherein the pH value is 6.8, and the mixture is sterilized at 121 ℃ for 20 minutes;13the culture was terminated when the C-labeled sucrose was exhausted. An LB culture medium is utilized for activation treatment, and the specific method comprises the following steps: taking out the serratia griffithii ACCC01695 from the slant of the test tube for preserving strains, transferring the strains to an LB culture medium, wherein the inoculation amount is 2 percent of volume, and carrying out inverted culture in a constant temperature box at 28 ℃ for 24 hours. The fermentation culture is aeration culture with air flux of 10L/min. The inoculation amount of the strain seed liquid in the fermentation medium is 6 percent by volume. 1/2 formula amount is added when preparing the fermentation medium13C labeling sucrose, monitoring during shaking culture13C the dosage of the cane sugar marked by the mark,13with additional remainder added in batches when sucrose marked C is exhausted13C labeling sucrose until all13C, marking that all the cane sugar is exhausted, and finishing the culture;13the number of times of adding the C-marked sucrose is 3, and the adding amount is the same each time. The centrifugation conditions were: centrifuging at 8000rpm/min for 15 min; the precipitation method comprises the following steps: and precipitating and collecting by using ammonium sulfate with the mass concentration of 50%.
The preparation method of the modified diatomite comprises the following steps: adding 1kg of sodium dodecyl benzene sulfonate into 5 times of water by weight, stirring for dissolving, then stirring and heating to 70 ℃, adding 2kg of activated diatomite, stirring for 3 hours under the nitrogen atmosphere, filtering, washing with water, and drying for 12 hours at 60 ℃ to obtain the sodium dodecyl benzene sulfonate. The activated diatomite is obtained by adding diatomite into water with the weight 5 times that of the diatomite, treating the diatomite for 3 minutes by using 800W microwave, centrifuging the diatomite and drying the diatomite for 2 hours at 120 ℃.
The preparation method of the underwater material comprises the following steps: pouring 1kg of polymeric aluminum ferric silicate, 0.1kg of cerium sulfate, 3kg of sodium humate and 0.3kg of Serratia grisea metabolite into 15kg of absolute ethyl alcohol, carrying out first ultrasonic oscillation, fully ball-milling, then adding 8kg of modified diatomite, carrying out second ultrasonic oscillation, carrying out microwave treatment, carrying out third ultrasonic oscillation, and naturally volatilizing the ethyl alcohol to obtain the underwater material. The treatment time for the first ultrasonic oscillation, the second ultrasonic oscillation, and the third ultrasonic oscillation was 30 minutes, 10 minutes, and 30 minutes, respectively. The process conditions of the microwave treatment are as follows: microwave treatment at 600W for 3 minutes.
In the step (1), a circular glass fiber reinforced plastic cultivation jar with the diameter of 2m and the depth of 0.8m is selected as a cultivation container.
In the step (2), the culture container is disinfected 1 day ahead (disinfected by 10ppm potassium permanganate) and then injected into non-polluted well water, and the water depth is 0.6 m.
The specific method of the step (3) is as follows: firstly, erecting a net cage in a culture container, selecting and placing the copperfish water flower seedlings with 4-day round mouths and film outlet into the net cage for culture, wherein the density is 3000 tails/m3Feeding the fries for 4 times every day at the temperature of 18 ℃, feeding 2 g/ten thousand of fairy bugs for 1-10 days, and feeding 4 g/ten thousand of fairy bugs for 11-20 days; after 10 days, the fry is diluted and transferred from the net cage to a culture container for direct culture, and the density is 1000 tails/m3Feeding after 20 days to reach 2cm, disinfecting with chopped tubificidae, and 30 days later, diluting the fry with density of 500 tails/m3After 60 days, the height reaches about 4cm, and then, adopting soft-shelled turtle materials and tubificidae according to the mass ratio of 1: 2 mixing and making dough for feeding. The aperture of the net cage is smaller than 1.0mm, and the size of the net cage is 1.4m 0.6 m. Feeding for 4 times per day at 8:00, 12:00, 18:00, 24:00 times. The size of the chopped tubificidae is less than 1mm, and the tubificidae is disinfected by soaking in 5ppm povidone iodine for 30 minutes; the protein content of the soft-shelled turtle material is more than 45 percent.
Example 2
A high-survival-rate cultivation method for fries of cupfish with round mouth comprises the following steps:
(1) selecting a culture container;
(2) water quality management: paving a layer of water bottom material at the water bottom, taking non-polluted well water as a culture water source for micro-flowing water culture, and aerating all day long by adopting an oxygenation pump;
(3) stocking the fry, and feeding with different baits at different stages;
the water bottom material is obtained by loading polymeric aluminum ferric silicate, cerium sulfate, sodium humate and a serratia grignard metabolite on modified diatomite, wherein the serratia grignard metabolite is obtained by activating, inoculating and fermenting serratia grignard ACCC 01695. The thickness of the bed material was 8 cm.
The preparation method of the metabolic product of the serratia griffithii comprises the following steps: activating the serratia griffithii ACCC01695, inoculating into a seed culture medium, and culturing at 31 deg.C for 15 hr to obtain strain seed solution; inoculating strain seed liquid into a fermentation culture medium, carrying out shaking culture at 31 ℃ and 200rpm/min, centrifuging, collecting fermentation supernatant, and precipitating to obtain a serratia griffithii metabolite; wherein, the formula of the seed culture medium is as follows: 12g of casein peptone, 9g of soluble starch, 8g of dextrin, 5g of sodium chloride, 4g of ammonium acetate and FeSO40.3g，CaCl24g, 1100mL of distilled water, 7.2 pH and 30 minutes of sterilization at 121 ℃; the formula of the fermentation medium is as follows: 6g of yeast extract is prepared by mixing 6g of yeast extract,13c-labeled sucrose 0.3g, MgSO4·7H2O 8g，NaNO38g, KCl12g and 1100mL of distilled water, wherein the pH value is 7.2, and the mixture is sterilized at 121 ℃ for 30 minutes;13the culture was terminated when the C-labeled sucrose was exhausted. An LB culture medium is utilized for activation treatment, and the specific method comprises the following steps: taking out the serratia griffithii ACCC01695 from the slant of the test tube for preserving strains, transferring the strains to an LB culture medium, wherein the inoculation amount is 3 percent by volume, and carrying out inverted culture in a constant temperature box at 28 ℃ for 72 hours. The fermentation culture is aeration culture, and the air flux is 12L/min. The inoculation amount of the strain seed liquid in the fermentation medium is 8 percent by volume. 1/2 formula amount is added when preparing the fermentation medium13C labeling sucrose, monitoring during shaking culture13C the dosage of the cane sugar marked by the mark,13with additional remainder added in batches when sucrose marked C is exhausted13C labeling sucrose until all13C, marking that all the cane sugar is exhausted, and finishing the culture;13the number of times of adding the C-marked sucrose is 5, and the adding amount is the same each time. The centrifugation conditions were: centrifuging at 12000rpm/min for 20 min; the precipitation method comprises the following steps: and precipitating and collecting by using ammonium sulfate with the mass concentration of 80%.
The preparation method of the modified diatomite comprises the following steps: adding 1kg of sodium dodecyl benzene sulfonate into 8 times of water by weight, stirring for dissolving, then stirring and heating to 80 ℃, adding 3kg of activated diatomite, stirring for 4 hours under nitrogen atmosphere, filtering, washing with water, and drying for 18 hours at 70 ℃ to obtain the sodium dodecyl benzene sulfonate. The activated diatomite is prepared by adding diatomite into 6 times of water by weight, treating for 5 minutes by 1000W microwave, centrifuging, and drying for 3 hours at 130 ℃.
The preparation method of the underwater material comprises the following steps: pouring 1kg of polymeric aluminum ferric silicate, 0.2kg of cerium sulfate, 4kg of sodium humate and 0.4kg of Serratia grisea metabolite into 20kg of absolute ethyl alcohol, carrying out first ultrasonic oscillation, fully ball-milling, then adding 10kg of modified diatomite, carrying out second ultrasonic oscillation, carrying out microwave treatment, carrying out third ultrasonic oscillation, and naturally volatilizing the ethyl alcohol to obtain the underwater material. The treatment time for the first ultrasonic oscillation, the second ultrasonic oscillation, and the third ultrasonic oscillation was 40 minutes, 15 minutes, and 40 minutes, respectively. The process conditions of the microwave treatment are as follows: and treating with 800W microwave for 5 minutes.
In the step (1), a circular glass fiber reinforced plastic cultivation jar with the diameter of 2m and the depth of 1m is selected as a cultivation container.
In the step (2), the culture container is disinfected 2 days in advance (disinfected by 10ppm potassium permanganate) and then injected into non-polluted well water, and the water depth is 0.8 m.
The specific method of the step (3) is as follows: firstly, erecting a net cage in a culture container, selecting and placing the copperfish fries with 5-day round mouth as a film in the net cage for culture, wherein the density is 2000 tails/m3Feeding the fries for 4 times every day at the temperature of 22 ℃, feeding 3 g/ten thousand of fairy bugs in 1-10 days, and feeding 5 g/ten thousand of fairy bugs in 11-20 days; after 10 days, the fry is diluted and transferred from the net cage to a culture container for direct culture, and the density is 1500 tails/m3Feeding after 20 days to reach about 2cm, disinfecting by using chopped tubificidae, and after 30 days, thinning the fry with the density of 800 tails/m3After 60 days, the height reaches about 4cm, and then, adopting soft-shelled turtle materials and tubificidae according to the mass ratio of 1: 4 mixing the materials to prepare dough and feeding the dough. The aperture of the net cage is smaller than 1.0mm, and the size of the net cage is 1.4m 0.6 m. Feeding for 4 times per day at 8:00, 12:00, 18:00, 24:00 times. The size of the chopped tubificidae is less than 1mm, and the tubificidae is disinfected by soaking in 5ppm povidone iodine for 30 minutes; protein mass content of soft-shelled turtle feedGreater than 45%.
Example 3
A high-survival-rate cultivation method for fries of cupfish with round mouth comprises the following steps:
(1) selecting a culture container;
(2) water quality management: paving a layer of water bottom material at the water bottom, taking non-polluted well water as a culture water source for micro-flowing water culture, and aerating all day long by adopting an oxygenation pump;
(3) stocking the fry, and feeding with different baits at different stages;
the water bottom material is obtained by loading polymeric aluminum ferric silicate, cerium sulfate, sodium humate and a serratia grignard metabolite on modified diatomite, wherein the serratia grignard metabolite is obtained by activating, inoculating and fermenting serratia grignard ACCC 01695. The thickness of the bed material was 6 cm.
The preparation method of the metabolic product of the serratia griffithii comprises the following steps: activating the serratia griffithii ACCC01695, inoculating into a seed culture medium, and culturing at 30 deg.C for 14 hr to obtain strain seed solution; inoculating strain seed liquid into a fermentation culture medium, carrying out shaking culture at 29 ℃ and 190rpm/min, centrifuging, collecting fermentation supernatant, and precipitating to obtain a serratia griffithii metabolite; wherein, the formula of the seed culture medium is as follows: 10g of casein peptone, 8g of soluble starch, 6g of dextrin, 4g of sodium chloride, 3g of ammonium acetate and FeSO40.25g，CaCl23.5g, 1000mL of distilled water, pH 7, sterilized at 121 ℃ for 25 minutes; the formula of the fermentation medium is as follows: 5g of yeast extract is prepared by mixing 5g of yeast extract,13c-labeled sucrose 0.25g, MgSO4·7H2O 6g，NaNO37g, KCl 10g and 1000mL of distilled water, wherein the pH value is 7.1, and the mixture is sterilized at 121 ℃ for 24 minutes;13the culture was terminated when the C-labeled sucrose was exhausted. An LB culture medium is utilized for activation treatment, and the specific method comprises the following steps: taking out the serratia griffithii ACCC01695 from the slant of the test tube for preserving strains, transferring the strains to an LB culture medium, wherein the inoculation amount is 2.5 percent by volume, and carrying out inverted culture in a constant temperature box at 28 ℃ for 36 hours. The fermentation culture is aeration culture, and the air flux is 11L/min. The inoculation amount of the strain seed liquid in the fermentation medium is 7 percent by volume. 1/2 formula amount is added when preparing the fermentation medium13C labeling sucrose, monitoring during shaking culture13C the dosage of the cane sugar marked by the mark,13with additional remainder added in batches when sucrose marked C is exhausted13C labeling sucrose until all13C, marking that all the cane sugar is exhausted, and finishing the culture;13the number of times of adding the C-marked sucrose is 4, and the adding amount is the same each time. The centrifugation conditions were: centrifuging at 11000rpm/min for 18 minutes; the precipitation method comprises the following steps: and precipitating and collecting by using ammonium sulfate with the mass concentration of 70%.
The preparation method of the modified diatomite comprises the following steps: adding 1kg of sodium dodecyl benzene sulfonate into 6 times of water by weight, stirring for dissolving, then stirring and heating to 75 ℃, adding 2.5kg of activated diatomite, stirring for 3.5 hours under nitrogen atmosphere, filtering, washing with water, and drying for 15 hours at 65 ℃ to obtain the sodium dodecyl benzene sulfonate. The activated diatomite is obtained by adding diatomite into 5 times of water by weight, treating for 4 minutes by 900W microwave, centrifuging and drying for 2 hours at 125 ℃.
The preparation method of the underwater material comprises the following steps: pouring 1kg of polymeric aluminum ferric silicate, 0.15kg of cerium sulfate, 3.5kg of sodium humate and 0.35kg of Serratia grisea metabolite into 18kg of absolute ethyl alcohol, carrying out first ultrasonic oscillation, carrying out full ball milling, then adding 9kg of modified diatomite, carrying out second ultrasonic oscillation, carrying out microwave treatment, carrying out third ultrasonic oscillation, and naturally volatilizing the ethyl alcohol to obtain the underwater material. The treatment time for the first ultrasonic oscillation, the second ultrasonic oscillation, and the third ultrasonic oscillation was 35 minutes, 12 minutes, and 33 minutes, respectively. The process conditions of the microwave treatment are as follows: microwave treatment at 750W for 4 minutes.
In the step (1), a circular glass fiber reinforced plastic cultivation jar with the diameter of 2m and the depth of 0.9m is selected as a cultivation container.
In the step (2), the culture container is disinfected 2 days in advance (disinfected by 10ppm potassium permanganate) and then injected into non-polluted well water, and the water depth is 0.7 m.
The specific method of the step (3) is as follows: firstly, erecting a net cage in a culture container, selecting and placing the copperfish fries with 4-day round mouth and film in the net cage for culture, wherein the density is 2500 tails/m3Feeding the fries for 4 times every day at the temperature of 20 ℃, feeding the fairy shrimp 2.5 g/ten thousand tails every 1-10 days, and feeding the fairy shrimp 4.5 g/ten thousand tails every 11-20 days; after 10 days, the fry is diluted anddirectly culturing from net cage to culture container with density of 1200 tail/m3Feeding after 20 days to reach about 2cm, disinfecting by using chopped tubificidae, and after 30 days, thinning the fry with the density of 700 tails/m3After 60 days, the height reaches about 4cm, and then, adopting soft-shelled turtle materials and tubificidae according to the mass ratio of 1: 3 mixing the materials to prepare dough and feeding the dough. The aperture of the net cage is smaller than 1.0mm, and the size of the net cage is 1.4m 0.6 m. Feeding for 4 times per day at 8:00, 12:00, 18:00, 24:00 times. The size of the chopped tubificidae is less than 1mm, and the tubificidae is disinfected by soaking in 5ppm povidone iodine for 30 minutes; the protein content of the soft-shelled turtle material is more than 45 percent.
Comparative example 1
A high-survival-rate cultivation method for fries of cupfish with round mouth comprises the following steps:
(1) selecting a culture container;
(2) water quality management: paving a layer of water bottom material at the water bottom, taking non-polluted well water as a culture water source for micro-flowing water culture, and aerating all day long by adopting an oxygenation pump;
(3) stocking the fry, and feeding with different baits at different stages;
the water bottom material is obtained by loading polymeric aluminum ferric silicate, cerium sulfate, sodium humate and serratia marcescens metabolites on modified diatomite, wherein the serratia marcescens metabolites are obtained by activating, inoculating and fermenting serratia marcescens ATCC 14756. The thickness of the bed material was 6 cm.
The preparation method of the metabolite of Serratia marcescens comprises the following steps: activating serratia marcescens ATCC14756, inoculating the activated serratia marcescens ATCC14756 into a seed culture medium, and culturing for 14 hours at 30 ℃ to obtain a strain seed solution; inoculating strain seed liquid into a fermentation culture medium, carrying out shaking culture at 29 ℃ and 190rpm/min, centrifuging, collecting fermentation supernatant, and precipitating to obtain a serratia marcescens metabolite; wherein, the formula of the seed culture medium is as follows: 10g of casein peptone, 8g of soluble starch, 6g of dextrin, 4g of sodium chloride, 3g of ammonium acetate and FeSO40.25g，CaCl23.5g, 1000mL of distilled water, pH 7, sterilized at 121 ℃ for 25 minutes; the formula of the fermentation medium is as follows: 5g of yeast extract is prepared by mixing 5g of yeast extract,13c-labeled sucrose 0.25g, MgSO4·7H2O 6g，NaNO37g, KCl 10g and 1000mL of distilled water, wherein the pH value is 7.1, and the mixture is sterilized at 121 ℃ for 24 minutes;13the culture was terminated when the C-labeled sucrose was exhausted. An LB culture medium is utilized for activation treatment, and the specific method comprises the following steps: the serratia marcescens ATCC14756 is taken out from the inclined plane of the test tube of the preserved strain and transferred to an LB culture medium, the inoculation amount is 2.5 percent of volume percent, and the inverted culture is carried out in a constant temperature box at 28 ℃ for 36 hours. The fermentation culture is aeration culture, and the air flux is 11L/min. The inoculation amount of the strain seed liquid in the fermentation medium is 7 percent by volume. 1/2 formula amount is added when preparing the fermentation medium13C labeling sucrose, monitoring during shaking culture13C the dosage of the cane sugar marked by the mark,13with additional remainder added in batches when sucrose marked C is exhausted13C labeling sucrose until all13C, marking that all the cane sugar is exhausted, and finishing the culture;13the number of times of adding the C-marked sucrose is 4, and the adding amount is the same each time. The centrifugation conditions were: centrifuging at 11000rpm/min for 18 minutes; the precipitation method comprises the following steps: and precipitating and collecting by using ammonium sulfate with the mass concentration of 70%.
The preparation method of the modified diatomite comprises the following steps: adding 1kg of sodium dodecyl benzene sulfonate into 6 times of water by weight, stirring for dissolving, then stirring and heating to 75 ℃, adding 2.5kg of activated diatomite, stirring for 3.5 hours under nitrogen atmosphere, filtering, washing with water, and drying for 15 hours at 65 ℃ to obtain the sodium dodecyl benzene sulfonate. The activated diatomite is obtained by adding diatomite into 5 times of water by weight, treating for 4 minutes by 900W microwave, centrifuging and drying for 2 hours at 125 ℃.
The preparation method of the underwater material comprises the following steps: pouring 1kg of polymeric aluminum ferric silicate, 0.15kg of cerium sulfate, 3.5kg of sodium humate and 0.35kg of Serratia grisea metabolite into 18kg of absolute ethyl alcohol, carrying out first ultrasonic oscillation, carrying out full ball milling, then adding 9kg of modified diatomite, carrying out second ultrasonic oscillation, carrying out microwave treatment, carrying out third ultrasonic oscillation, and naturally volatilizing the ethyl alcohol to obtain the underwater material. The treatment time for the first ultrasonic oscillation, the second ultrasonic oscillation, and the third ultrasonic oscillation was 35 minutes, 12 minutes, and 33 minutes, respectively. The process conditions of the microwave treatment are as follows: microwave treatment at 750W for 4 minutes.
In the step (1), a circular glass fiber reinforced plastic cultivation jar with the diameter of 2m and the depth of 0.9m is selected as a cultivation container.
In the step (2), the culture container is disinfected 2 days in advance (disinfected by 10ppm potassium permanganate) and then injected into non-polluted well water, and the water depth is 0.7 m.
The specific method of the step (3) is as follows: firstly, erecting a net cage in a culture container, selecting and placing the copperfish fries with 4-day round mouth and film in the net cage for culture, wherein the density is 2500 tails/m3Feeding the fries for 4 times every day at the temperature of 20 ℃, feeding the fairy shrimp 2.5 g/ten thousand tails every 1-10 days, and feeding the fairy shrimp 4.5 g/ten thousand tails every 11-20 days; after 10 days, the fry is diluted and transferred from the net cage to a culture container for direct culture, and the density is 1200 tails/m3Feeding after 20 days to reach about 2cm, disinfecting by using chopped tubificidae, and after 30 days, thinning the fry with the density of 700 tails/m3After 60 days, the height reaches about 4cm, and then, adopting soft-shelled turtle materials and tubificidae according to the mass ratio of 1: 3 mixing the materials to prepare dough and feeding the dough. The aperture of the net cage is smaller than 1.0mm, and the size of the net cage is 1.4m 0.6 m. Feeding for 4 times per day at 8:00, 12:00, 18:00, 24:00 times. The size of the chopped tubificidae is less than 1mm, and the tubificidae is disinfected by soaking in 5ppm povidone iodine for 30 minutes; the protein content of the soft-shelled turtle material is more than 45 percent.
Comparative example 2
A high-survival-rate cultivation method for fries of cupfish with round mouth comprises the following steps:
(1) selecting a culture container;
(2) water quality management: paving a layer of water bottom material at the water bottom, taking non-polluted well water as a culture water source for micro-flowing water culture, and aerating all day long by adopting an oxygenation pump;
(3) stocking the fry, and feeding with different baits at different stages;
the water bottom material is obtained by loading polymeric aluminum ferric silicate, sodium humate and a serratia grignard metabolite on modified diatomite, wherein the serratia grignard metabolite is obtained by activating, inoculating and fermenting serratia grignard ACCC 01695. The thickness of the bed material was 6 cm.
The preparation method of the metabolic product of the serratia griffithii comprises the following steps: activating the serratia griffithii ACCC01695, inoculating in seed culture medium,culturing at 30 deg.C for 14 hr to obtain strain seed solution; inoculating strain seed liquid into a fermentation culture medium, carrying out shaking culture at 29 ℃ and 190rpm/min, centrifuging, collecting fermentation supernatant, and precipitating to obtain a serratia griffithii metabolite; wherein, the formula of the seed culture medium is as follows: 10g of casein peptone, 8g of soluble starch, 6g of dextrin, 4g of sodium chloride, 3g of ammonium acetate and FeSO40.25g，CaCl23.5g, 1000mL of distilled water, pH 7, sterilized at 121 ℃ for 25 minutes; the formula of the fermentation medium is as follows: 5g of yeast extract is prepared by mixing 5g of yeast extract,13c-labeled sucrose 0.25g, MgSO4·7H2O 6g，NaNO37g, KCl 10g and 1000mL of distilled water, wherein the pH value is 7.1, and the mixture is sterilized at 121 ℃ for 24 minutes;13the culture was terminated when the C-labeled sucrose was exhausted. An LB culture medium is utilized for activation treatment, and the specific method comprises the following steps: taking out the serratia griffithii ACCC01695 from the slant of the test tube for preserving strains, transferring the strains to an LB culture medium, wherein the inoculation amount is 2.5 percent by volume, and carrying out inverted culture in a constant temperature box at 28 ℃ for 36 hours. The fermentation culture is aeration culture, and the air flux is 11L/min. The inoculation amount of the strain seed liquid in the fermentation medium is 7 percent by volume. 1/2 formula amount is added when preparing the fermentation medium13C labeling sucrose, monitoring during shaking culture13C the dosage of the cane sugar marked by the mark,13with additional remainder added in batches when sucrose marked C is exhausted13C labeling sucrose until all13C, marking that all the cane sugar is exhausted, and finishing the culture;13the number of times of adding the C-marked sucrose is 4, and the adding amount is the same each time. The centrifugation conditions were: centrifuging at 11000rpm/min for 18 minutes; the precipitation method comprises the following steps: and precipitating and collecting by using ammonium sulfate with the mass concentration of 70%.
The preparation method of the modified diatomite comprises the following steps: adding 1kg of sodium dodecyl benzene sulfonate into 6 times of water by weight, stirring for dissolving, then stirring and heating to 75 ℃, adding 2.5kg of activated diatomite, stirring for 3.5 hours under nitrogen atmosphere, filtering, washing with water, and drying for 15 hours at 65 ℃ to obtain the sodium dodecyl benzene sulfonate. The activated diatomite is obtained by adding diatomite into 5 times of water by weight, treating for 4 minutes by 900W microwave, centrifuging and drying for 2 hours at 125 ℃.
The preparation method of the underwater material comprises the following steps: pouring 1kg of polymeric aluminum ferric silicate, 3.5kg of sodium humate and 0.35kg of serratia grignard metabolite into 18kg of absolute ethyl alcohol, carrying out ultrasonic oscillation for the first time, carrying out full ball milling, then adding 9kg of modified diatomite, carrying out ultrasonic oscillation for the second time, carrying out microwave treatment, carrying out ultrasonic oscillation for the third time, and naturally volatilizing the ethyl alcohol to obtain the underwater material. The treatment time for the first ultrasonic oscillation, the second ultrasonic oscillation, and the third ultrasonic oscillation was 35 minutes, 12 minutes, and 33 minutes, respectively. The process conditions of the microwave treatment are as follows: microwave treatment at 750W for 4 minutes.
In the step (1), a circular glass fiber reinforced plastic cultivation jar with the diameter of 2m and the depth of 0.9m is selected as a cultivation container.
In the step (2), the culture container is disinfected 2 days in advance (disinfected by 10ppm potassium permanganate) and then injected into non-polluted well water, and the water depth is 0.7 m.
The specific method of the step (3) is as follows: firstly, erecting a net cage in a culture container, selecting and placing the copperfish fries with 4-day round mouth and film in the net cage for culture, wherein the density is 2500 tails/m3Feeding the fries for 4 times every day at the temperature of 20 ℃, feeding the fairy shrimp 2.5 g/ten thousand tails every 1-10 days, and feeding the fairy shrimp 4.5 g/ten thousand tails every 11-20 days; after 10 days, the fry is diluted and transferred from the net cage to a culture container for direct culture, and the density is 1200 tails/m3Feeding after 20 days to reach about 2cm, disinfecting by using chopped tubificidae, and after 30 days, thinning the fry with the density of 700 tails/m3After 60 days, the height reaches about 4cm, and then, adopting soft-shelled turtle materials and tubificidae according to the mass ratio of 1: 3 mixing the materials to prepare dough and feeding the dough. The aperture of the net cage is smaller than 1.0mm, and the size of the net cage is 1.4m 0.6 m. Feeding for 4 times per day at 8:00, 12:00, 18:00, 24:00 times. The size of the chopped tubificidae is less than 1mm, and the tubificidae is disinfected by soaking in 5ppm povidone iodine for 30 minutes; the protein content of the soft-shelled turtle material is more than 45 percent.
Comparative example 3
A high-survival-rate cultivation method for fries of cupfish with round mouth comprises the following steps:
(1) selecting a culture container;
(2) water quality management: culturing in micro-flowing water by using pollution-free well water as a culture water source, and aerating all day by using an oxygenation pump;
(3) fry are stocked and fed with different baits in different stages.
In the step (1), a circular glass fiber reinforced plastic cultivation jar with the diameter of 2m and the depth of 0.9m is selected as a cultivation container.
In the step (2), the culture container is disinfected 2 days in advance (disinfected by 10ppm potassium permanganate) and then injected into non-polluted well water, and the water depth is 0.7 m.
The specific method of the step (3) is as follows: firstly, erecting a net cage in a culture container, selecting and placing the copperfish fries with 4-day round mouth and film in the net cage for culture, wherein the density is 2500 tails/m3Feeding the fries for 4 times every day at the temperature of 20 ℃, feeding the fairy shrimp 2.5 g/ten thousand tails every 1-10 days, and feeding the fairy shrimp 4.5 g/ten thousand tails every 11-20 days; after 10 days, the fry is diluted and transferred from the net cage to a culture container for direct culture, and the density is 1200 tails/m3Feeding after 20 days to reach about 2cm, disinfecting by using chopped tubificidae, and after 30 days, thinning the fry with the density of 700 tails/m3After 60 days, the height reaches about 4cm, and then, adopting soft-shelled turtle materials and tubificidae according to the mass ratio of 1: 3 mixing the materials to prepare dough and feeding the dough. The aperture of the net cage is smaller than 1.0mm, and the size of the net cage is 1.4m 0.6 m. Feeding for 4 times per day at 8:00, 12:00, 18:00, 24:00 times. The size of the chopped tubificidae is less than 1mm, and the tubificidae is disinfected by soaking in 5ppm povidone iodine for 30 minutes; the protein content of the soft-shelled turtle material is more than 45 percent.
Test examples
The method of examples 1 to 3 or comparative examples 1 to 3 was used to simultaneously breed the round-mouth coppers during the 7 th month to the 6 th month in 2018, respectively, the stocking fraction (according to the predetermined stocking density) and the harvest fraction were counted, and the survival rate was calculated, with the results shown in table 1.
TABLE 1 comparison of incubation conditions
| Stocking mantissa (Tail) | Harvest mantissa (tail) | Survival rate (%) | |
| Example 1 | 5652 | 5387 | 95.3 |
| Example 2 | 5024 | 4798 | 95.5 |
| Example 3 | 5495 | 5319 | 96.8 |
| Comparative example 1 | 5495 | 4698 | 85.5 |
| Comparative example 2 | 5495 | 4407 | 80.2 |
| Comparative example 3 | 5495 | 3313 | 60.3 |
As is clear from Table 1, the method of examples 1 to 3 exhibited a high survival rate of the cultured cuprins. In comparative example 1, serratia marcescens ATCC14756 (purchased from Shanghai Noro Biotech Co., Ltd.) is used for replacing serratia marcescens ACCC01695, cerium sulfate is omitted when a water bottom material is prepared in comparative example 2, a water bottom material is not laid in comparative example 3, the survival rate of round-mouth copper fish is obviously reduced, the serratia marcescens metabolite has specificity on the improvement of immunity of round-mouth copper fish, the metabolite obtained by replacing similar strains has obviously lower improvement degree of immunity, and the cerium sulfate in the water bottom material and polymeric aluminum ferric silicate and the like synergistically purify water, so that the survival rate of round-mouth copper fish is improved.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (5)
1. A high-survival-rate cultivation method for fries of cupfish with round mouth is characterized by comprising the following steps:
(1) selecting a culture container;
(2) water quality management: paving a layer of water bottom material at the water bottom, taking non-polluted well water as a culture water source for micro-flowing water culture, and aerating all day long by adopting an oxygenation pump;
(3) stocking the fry, and feeding with different baits at different stages;
the water bottom material is obtained by loading polymeric aluminum ferric silicate, cerium sulfate, sodium humate and a serratia grignard metabolite on modified diatomite, wherein the serratia grignard metabolite is obtained by activating, inoculating and fermenting serratia grignard ACCC 01695;
the preparation method of the modified diatomite comprises the following steps: adding 1 part by weight of sodium dodecyl benzene sulfonate into 5-8 times of water by weight, stirring for dissolving, then stirring and heating to 70-80 ℃, adding 2-3 parts of activated diatomite, stirring for 3-4 hours under nitrogen atmosphere, filtering, washing with water, and drying to obtain the sodium dodecyl benzene sulfonate-containing composite material;
the preparation method of the underwater material comprises the following steps: pouring 1 part of polymeric aluminum ferric silicate, 0.1-0.2 part of cerium sulfate, 3-4 parts of sodium humate and 0.3-0.4 part of a metabolic product of serratia grignard into 15-20 parts of absolute ethyl alcohol, performing first ultrasonic oscillation and full ball milling, then adding 8-10 parts of modified diatomite, performing second ultrasonic oscillation, performing microwave treatment, performing third ultrasonic oscillation, and naturally volatilizing ethanol to obtain the underwater material;
wherein, the specific method of the step (3) is as follows: firstly, erecting a net cage in a culture container, selecting a round-mouth copperfish water flower seedling with a film outlet of 4-5 days, and putting the round-mouth copperfish water flower seedling into the net cage for culture, wherein the density is 2000-3000 tails/m3Feeding the fries for 4 times every day at the temperature of 18-22 ℃, feeding 2-3 g/ten thousand of fairy bugs in 1-10 days, and feeding 4-5 g/ten thousand of fairy bugs in 11-20 days; after 10 days, thinning the fry and transferring the fry from the net cage to a culture container for direct culture, wherein the density is 1000-1500 tails/m3Feeding after 20 days to about 2cm, disinfecting by adopting chopped tubificidae, and after 30 days, thinning the fry with the density of 500-800 tails/m3After 60 days, the height reaches about 4cm, and then, adopting soft-shelled turtle materials and tubificidae according to the mass ratio of 1: 2-4, mixing, making dough and feeding;
the preparation method of the metabolic product of the serratia griffithii comprises the following steps: activating the serratia griffithii ACCC01695, inoculating the activated serratia griffithii ACCC01695 into a seed culture medium, and culturing at 28-31 ℃ for 12-15 hours to obtain a strain seed solution; inoculating strain seed liquid into a fermentation culture medium, carrying out shaking culture at 27-31 ℃ and 180-200 rpm/min, centrifuging, collecting fermentation supernatant, and precipitating to obtain a serratia griffithii metabolite;wherein, the formula of the seed culture medium is as follows: 8-12 g of casein peptone, 6-9 g of soluble starch, 5-8 g of dextrin, 3-5 g of sodium chloride, 2-4 g of ammonium acetate and FeSO4 0.2～0.3g，CaCl23-4 g of distilled water, 900-1100 mL of distilled water, 6.8-7.2 of pH, and sterilizing for 20-30 minutes at 121 ℃; the formula of the fermentation medium is as follows: 4-6 g of yeast extract,13c-labeled sucrose 0.2-0.3 g, MgSO4 & 7H25-8 g of O, 35-8 g of NaNO, 8-12 g of KCl and 900-1100 mL of distilled water, wherein the pH value is 6.8-7.2, and the mixture is sterilized at 121 ℃ for 20-30 minutes;13the culture was terminated when the C-labeled sucrose was exhausted.
2. The cultivation method as claimed in claim 1, wherein the thickness of the underwater material is 5 to 8 cm.
3. The cultivation method according to claim 1, wherein in the step (1), a circular glass fiber reinforced plastic cultivation jar with a diameter of 2m and a depth of 0.8-1 m is selected as the cultivation container.
4. The method of claim 1, wherein the mesh cage has a pore size of less than 1.0mm, and the mesh cage has a size of 1.4m by 0.6 m.
5. The method of claim 1, wherein the size of the chopped tubificidae is less than 1mm, and the tubificidae is disinfected by soaking in 5ppm povidone-iodine for 30 minutes; the protein content of the soft-shelled turtle material is more than 45 percent.
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