CN104255626B - A kind of raising is survived the winter period cage-farming immunity and the method for disease resistance - Google Patents
A kind of raising is survived the winter period cage-farming immunity and the method for disease resistance Download PDFInfo
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Classifications
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- 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
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Farming Of Fish And Shellfish (AREA)
- Fodder In General (AREA)
Abstract
The invention discloses a kind of method of cage-farming immunity and disease resistance during raising is survived the winter, step: A, test process: matched group is period glitch-free cage culture of surviving the winter; Process group drags net cage once weekly for period of surviving the winter, and makes Carassius aurutus gibelio expose in atmosphere, then puts back in water by fish and net cage, the Fish that survive the winter in net cage are carried out primary air exposure and fish for stimulation, man-made feeds of not throwing something and feeding; B, winter test: record water temperature, natural lighting; C, before winter test, test fish carries out fortification in net cage system, and the enriched nutritive phase is one month, and all of cultured fishes are not thrown something and fed artifical compound feed; D, challenge test; E, winter test terminate, and put back to and continue in former net cage to raise after adopting lumbar injection Aeromonas hydrophila liquid. Easy to implement the method, easy and simple to handle, enhance the immunity of cage-farming and the resistivity to pathogenic bacteria, significantly improve the survival rate after the infection of marine fishes pathogenic bacteria that survives the winter.
Description
Technical field
The invention belongs to technical field of aquaculture, be more particularly to a kind of method that can improve survive the winter period cage-farming immunity and disease resistance.
Background technology
Fish are a kind of poikilothermal animals, and body temperature changes along with the change of external environment. Simultaneously, Fish are more sensitive to the variations in temperature of external aqueous environment, Fish are also variant in different temperatures scope to the sensitivity of ambient temperature, generally, Fish belong to " chill sensitive ", namely sensitivity habituation when increased response and temperature raise when temperature reduces (Yi Xianghua etc., Carnis Pseudosciaenae survives the winter experiment. aquatic science and technology information, 1998,25 (6): 261-264). For temperate water fishes, winter is a harsh stage, and low water temperature causes the enhancing of temperature sensitivity, to overcome the impact that low temperature brings on the one hand, to stand food deprivation on the other hand and even stop eating the various undesirable changes caused. The pressure that period Fish face it can be said that survive the winter is dual. Simultaneously, due to the impact of artificial limitation (such as artificial cultivation) etc., most temperate water fisheses all live in a certain specific region in life, and winter is also a very crucial period, or strengthen adaptability to survive, or exceed tolerance limit and large quantities of death. Therefore, strengthen winter management, improve fingerling overwintering survival rate and production and the stable and high yields in the coming year are had very important significance.
The kind that Carassius auratus, channel catfish and Ctenopharyngodon idellus etc. widely cultivate at present all belongs to temperate water fishes, just stopping ingesting or seldom ingesting when water temperature is lower than 10 DEG C, the activity of Fish weakens, and stops growing, metabolism slows down, and its metabolic energy derives from the nutrient substance of himself internal accumulation.In such a situa-tion, fish should maintain basic activity and consume supplementing of energy substance, nutrition enough less than the external world again and energy matter. Therefore, Fish are through spring winter to next year, and body weight mostly alleviates, the immunity degradation of fish itself, and premunition is weak, it is easy to by antibacterial, fungus and parasitic infection and invasion and attack, and the mortality rate of the Fish that cause surviving the winter significantly rises.
Survive the winter period, Fish depend on the energy matter of storage to meet analytic metabolism demand (Post etc., Size-dependentoverwintermortalityofyoung-of-the-yearyell owpecrh (percaflavescens): laboratory, insituenclosure, andfieldexperiments.CanadianJournalofFisheriesandAquatic Sciences, 1989:46,1958-1968). If overwintering death simply because energy decline cause, then thermokinetic theory think low temperature time should have low mortality rate because low temperature makes metabolic rate decline, energy expenditure is less. But, it was recently reported that white bass (Moroneamericana) overwintering mortality up to 63% and consume less energy during than 4 DEG C when 2.5 DEG C, describe the participation of energy expenditure other factors outer. The destruction of hunger, water balance and ion-select electrode is possible factor.
Under starvation, Metabolism of Fishes level will be decreased obviously. Research find the metabolism after hungry 15d of perch juvenile fish have dropped about 45%(Mehner etc., Energeticsandmetaboliccorrelatesofstarvationinjuvenilepe rch (Percafluviatilis) .JournalofFishBiology, 1994:45,325-333). Speckle grunts (pomadasyscommersonni) consumption rate have dropped 34%(DuPreez etc. after hungry 5d, Oxygenconsumptionofashallowwaterteleost, thespottedgrunter, Pomadasyscommersonni (Lac �� p �� de, 1802) .ComparativeBiochemistryandPhysiologyA, 1986:84,61-70). Ctenopharyngodon idellus finds, the metabolic rate after hungry 35d significantly lower than under normal condition measured value (Cui Yibo etc., the relation of the metabolic rate of Ctenopharyngodon idellus and nitrogen excretion and gas weight under starvation. aquatile journal, 1993,17:375-376).
Fish stop when low temperature ingesting or seldom ingesting, and the most significantly change is weight loss. Because can not get supplementing of exogenous diet, and carrying out the energy expenditure of necessity for maintaining basic existence, therefore muscle and some viscera tissue weight there occurs change, and nearly all hungry Fish that survive the winter all occur in that being remarkably decreased of body weight regulating liver-QI weight simultaneously. The period hungry body weight to fish of surviving the winter creates significant impact. It is reported that, the channel catfish ingested continuously of period of surviving the winter ingest with interruption (December, January and February do not throw something and feed) channel catfish compared with, body weight does not have significant change, and the fish of hunger is by comparison, and weight differences is obvious. The Ctenopharyngodon idellus survived the winter also has similar result, and the less Grass Carp Juveniles moon rate of body weight gain of throwing something and feeding still reaches 2.3%, and under equal conditions, larva starvation group has become thin 3.43%. Ctenopharyngodon idellus is after hunger, and coefficient of condition, dirty body ratio, liver body ratio significantly reduce. Along with the prolongation of starvation time, the moisture in Ctenopharyngodon idellus muscle is gradually increased, and Oil content and Protein content presents reduction trend, moisture and protein content and presents the trend that diversity increases and reduces respectively.
Surviving the winter period, the change of Fish weight loss is also created obvious effect by water temperature.The low temperature hunger test in Europe perch (Dicentrarchuslabrax) shows, weight loss is more significantly and consistent with the adjustment of temperature at hungry first week. At the temperature of 3 DEG C, 7 DEG C, 8 DEG C, the amplitude of Europe perch weight loss is different, and respectively 37%, 34% and 42%, it is thought that the cost to low-temperature adaptation, because the temperature that Europe perch stops eating is 7 DEG C, below or above such temperature, more energy will be consumed and deacclimatize. And, the body weight of the European perch that the domestication water temperature before test is stopped eating on surviving the winter also has impact. The fish that acclimation temperature is high lose weight degree bigger (Pastoureaud, Influenceofstarvationatlowtemperaturesonutilizationofene rgyreserves, appetiterecoveryandgrowthcharacterinseabass,Dicentrarchuslabrax.Aquaculture, 1991,99:167 178).
Fish are when hungry, in order to make metaboilic level reduce, and can by suppressing the hemopoietic function of fish body and reducing oxygen demand [50]. Research shows, along with the prolongation of starvation time, Wintering Period Pelteobagrus fulvidraco serum cholesterol, total protein, is gradually reduced from albumen and globulin content. After finding hungry 28 days in Megalobrama amblycephala, each body weight group triglyceride concentration, blood glucose and total cholesterol concentration are all remarkably decreased; In serum, to compare triglyceride and T-CHOL more sensitive to hunger for blood sugar concentration. After research finds Nature enemy, Carassius aurutus gibelio Blood Glucose, triglyceride content significantly reduce, erythrocyte number, content of hemoglobin and cholesterol level first significantly reduce, go back up to matched group level subsequently, the quantity of leukocyte, the erythrocytic line of apsides, erythrocyte sedimentation rate and total protein have no significant change (Shen Wenying etc., the hungry impact on pond crucian carp blood constitutent and the development of ovary. zoological research, 2003,24 (6): 441-444).
Survive the winter period, the immunity of Fish and the resistance of disease is subject to very big impact. Under the stressors such as low water temperature and hunger stimulate, fish interior free yl can increase, serious harm body health, and body is in order to keep the dynamic equilibrium of interior free yl, it is necessary to removes too much free radical in time. Antioxidative Defense System is Free-radical ring opening polymerization important in body, and this defense system has enzymatic and non-enzymatic reaction two parts. Research finds along with the prolongation of starvation time, Procambrus clarkii (Procambarusclarkii) superoxide dismutase (SOD) and catalase (CAT) activity be all significantly reduce after first somewhat going up height, the immunity of shrimp is decreased obviously. Having been reported that discovery, hunger may result in Pelteobagrus fulvidraco nutrition supply not enough, the physiological function such as metaboilic level, immunity declines. First, hunger makes Pelteobagrus fulvidraco immune organ heavily decline. Secondly, NBT (NBT) positive cell number represents the phagocytic activity of phagocyte, neutrophilic granulocyte, macrophage, and as hungry Russia 12d, NBT positive cell number is remarkably decreased. And lysozyme activity in serum, hemolytic activity, antibacterial ability be remarkably decreased after hungry 18d (Sun Hongmei etc., the hungry impact on Pelteobagrus fulvidraco immune function. water conservancy related fisheries, 2006,26 (3): 80-81).
Hua Xueming etc. (2012) have studied the long-term hungry impact on Ctenopharyngodon idellus innate immune activity, find by the impact that hunger is coerced, Ctenopharyngodon idellus NKT sexual cell killing activity in spleen and head-kidney is substantially less than matched group, and not along with the prolongation generation significance of starvation time changes;Along with the prolongation of starvation time, in serum regulating liver-QI pancreas, lysozyme presents and first raises the trend reduced afterwards, and serum alkaline phosphatase is substantially less than matched group when hungry 15d, 45d, 60d; The alkaline phosphatase activities of larva starvation group, after hungry 30d, remains constant. As can be seen here, long hunger coerce reduce Ctenopharyngodon idellus immune level (Hua Xueming etc., the hungry impact on Ctenopharyngodon idellus nonspecific immunity level. zoology magazine, 2012,47 (2): 91-97).
Studies have reported that impact that is hungry and that throw something and feed on Carassius aurutus gibelio Blood Physiological Indexes and salivary lysozyme again. Research finds, hunger makes blood glucose and malonaldehyde (MDA) content pole significantly reduce, and with the prolongation of starvation time, SOD activity gradually rises after being remarkably decreased, acid phosphatase (ACP) activity is not made significant difference by Short-term starvation, and prolongation starvation time then first raises and is remarkably decreased afterwards further; After refeeding, Short-term starvation group blood sugar concentration, SOD and ACP activity all significantly rise, mid-term larva starvation group blood glucose, content of hemoglobin and ACP activity be gradually increasing, SOD activity and MDA content are remarkably decreased, long-term larva starvation group blood glucose and MDA content are substantially less than matched group, and ACP activity is gradually restored to control level. Result shows, throw something and feed again after Short-term starvation and can improve Carassius aurutus gibelio metaboilic level and nonspecific mental retardation (Dong Xuexing etc., throw something and feed again after hunger the impact on Carassius aurutus gibelio blood physiology and salivary lysozyme. China's agronomy circular, 2011,27 (23): 76-79).
As can be seen here, the double influence of low water temperature and hunger it is affected by period of surviving the winter most metabolism of temperate water fishes, physiology, biochemical indicator, immunity and the resistance etc. to disease, easily by antibacterial, fungus and parasitic infection and invasion and attack, the mortality rate of the Fish that cause surviving the winter significantly rises. And China's most area is in temperate zone, sub-temperate zone environment, the bottleneck problem that warm water cultured fishes ubiquity is survived the winter. Therefore, strengthening the aquaculture management during surviving the winter, the sound development of aquaculture is had very important significance by the overwintering survival rate improving cultured fishes.
At present, studies have reported that the immunity of Fish can be strengthened by the training repeated. Zhu Zhiming etc. (2011) have studied persistent movement take exercise lower different motion intensity to many squamas four must (Barbodesschwanenfeldi) impact of juvenile fish blood non-specific immunity. It was found that NBT positive cell quantity phase in an experiment and later stage in exercise group blood are all remarkably higher than matched group. Antalzyme activity and SOD enzyme activity also significantly raise after motion exercise. During the Serum bactericidal vigor phase in an experiment, exercise group is all remarkably higher than matched group, but is not significantly different between exercise group and matched group when testing latter stage. Test result indicate that, suitable motion exercise can strengthen many squamas four must juvenile fish non-specific immunity. Costas etc. (2011) it was also found that Senegal sole (Soleasenegalensis) expose every day in atmosphere 3 minutes, through two weeks repetitive stimulations train after, partial immunity index is significantly improved, to pathogenic bacteria (Photobacteriumdamselaesubsp.piscicida) resistivity be significantly enhanced (Costas etc., Dietaryarginineandrepeatedhandlingincreasediseaseresista nceandmodulateinnateimmunemechanismsofSenegalesesole (SoleasenegalensisKaup, 1858) .Fish&ShellfishImmunology, 2011,31:838-847).
Summary of the invention
It is an object of the invention to there are provided a kind of method of cage-farming immunity and disease resistance during raising is survived the winter, easy to implement the method, easy and simple to handle, improvement by the aquaculture management method of period of surviving the winter, can effectively enhance the immunity of cage-farming and the resistivity to pathogenic bacteria, significantly improve the survival rate after the infection of marine fishes pathogenic bacteria that survives the winter.
In order to realize above-mentioned purpose, the present invention adopts techniques below measure:
A kind of raising is survived the winter period cage-farming immunity and the method for disease resistance, the steps include:
A, whole survive the winter period, regularly drag net cage once weekly, after making the fish in net cage expose 4-6 minute in atmosphere, again fish and net cage are put back in water, with this, Fish that survive the winter in net cage carried out weekly air exposure and fish for stimulation, matched group has 3 tests parallel respectively with test process group, totally 6 mouthfuls of net cages. Whole surviving the winter period, net cage Fish are not thrown something and fed man-made feeds. Winter test: test in the Changjiang river therefore carry out in 6 mouthfuls of net cages (2m �� 2m �� 2m) in river course. During test, recording water temperature every day, the water temperature of December is 4 DEG C-10.5 DEG C, and the water temperature in January is 3.5 DEG C-8.0 DEG C; The water temperature in February is 5.5 DEG C-9.0 DEG C; The water temperature in March is 9.0 DEG C-14.0 DEG C. Illumination condition is natural lighting, and water pH value is 7.3-7.5. Regular air exposure and fish for the immunity stimulated for strengthening cage-farming of surviving the winter and to reduce the test effect of the mortality rate after bacteria infection very good, asks for an interview the effect described in embodiment 1 to 3. Described cage culture is pond net box cultivation, river course cage culture or cages in reservoir cultivation;
B, step A obtain the method for cultivation relevant with the frequency that the actual effect of pathogenic bacteria resistance was dragged with period of surviving the winter net cage and cultured fishes aerial open-assembly time to strengthening period cage-farming immunity of surviving the winter, it is typically in surviving the winter the operation of period 08:00-09:00 in the morning on every Mondays once, every time by fish together with net cage at air exposure 4-6 minute, drag net cage once by same procedure weekly;
C, relative to the Carassius aurutus gibelio (comparison fish) that whole period of surviving the winter is not subject to any interference, after period cage culture Carassius aurutus gibelio of surviving the winter being managed according to the method for the cultivation obtained in step A, immunity and the viral resistance of Carassius aurutus gibelio of surviving the winter can be significantly increased, mainly include Carassius aurutus gibelio survive the winter end time, catalase (CAT) vigor 86% of fish body blood plasma can be significantly improved, improve Plasma Total Antioxidant ability (T-AOC) 36.5%, improve blood plasma total number born (T-SOD) vigor 14.5%, significantly reduce lipid peroxidation product malonaldehyde (MDA) content 21.9% that can indirectly reflect cell injury in blood plasma,After infected by Aeromonas hydrophila fish body 168h, it is possible on the basis of comparison fish cumulative mortality 76.9%, reduce by 20.6% and reach the low actual of 56.3%; 6h, 24h and 72h after infected by Aeromonas hydrophila, blood plasma catalase (CAT) vigor has been respectively increased 181.5%, 56.8% and 37.1%; 6h and 24h after infected by Aeromonas hydrophila, Acute Coronary Syndrome (MPO) vigor has been respectively increased 33.3% and 117.6%; 72h and 168h after infected by Aeromonas hydrophila, Plasma Total Antioxidant ability (T-AOC) has been respectively increased 51.9% and 59.1%;72h after infected by Aeromonas hydrophila, Plasma MDA (MDA) content have dropped 40.6%; After infected by Aeromonas hydrophila, sharply stress falling after rise after the first notable rising of key index level of plasma cortisol of comparison fish, 6h after counteracting toxic substances reaches the highest (increasing 293.4%), and does not have significant change according to the Carassius aurutus gibelio cortisol levels after processing of step A after counteracting toxic substances.
Relative to the Carassius aurutus gibelio (comparison fish) that whole period of surviving the winter is not subject to any interference, according to the Carassius aurutus gibelio after processing of step A survive the winter heat stress proteins HSP70 and Ia interleukin I L11 in spleen when terminating, kidney and head-kidney, marrow sample differentiation factor MyD88 and Toll-like receptor TLR3 gene expression amount do not have significant change. But, 6h after infected by Aeromonas hydrophila, the gene expression amount of the heat stress proteins HSP70 of comparison fish spleen, kidney and head-kidney than improves 242.2%, 145.8% and 16.9 times before infecting respectively, then only than improves 142.0%, 99.9% and 1.6 times before infecting respectively according to the gene expression amount of the heat stress proteins HSP70 of the Carassius aurutus gibelio spleen after processing of step A, kidney and head-kidney; 24h after infected by Aeromonas hydrophila, the gene expression amount of the interleukin I L11 of comparison fish spleen, kidney and head-kidney than improves 2490.9 times, 57.8 times and 268.1 times before infecting respectively, then only improves 1381.5 times, 33.2 times and 19.5 times than before infecting respectively according to the gene expression amount of the interleukin I L11 of the Carassius aurutus gibelio spleen after processing of step A, kidney and head-kidney.
D, relative to the channel catfish (comparison fish) that whole period of surviving the winter is not subject to any interference, after period cage culture channel catfish of surviving the winter being managed according to the aquaculture management method obtained in step A, the immunity of channel catfish of surviving the winter can be significantly increased, mainly include channel catfish survive the winter end time, catalase (CAT) vigor 53.4% of fish body blood plasma can be significantly improved, significantly reduce lipid peroxidation product malonaldehyde (MDA) content 15.4% that can indirectly reflect cell injury in blood plasma, key index level of plasma cortisol stress significantly reduce 37.8%, 6h after infected by Aeromonas hydrophila fish body, 24h, 48h, 72h and 168h, blood plasma catalase (CAT) vigor has been respectively increased 64.7%, 65.6%, 79.3%, 48.9% and 27.4%, 24h, 48h and 72h after infected by Aeromonas hydrophila, Acute Coronary Syndrome (MPO) vigor has been respectively increased 39.8%, 13.3% and 32.5%, 6h, 24h and 48h after infected by Aeromonas hydrophila, Plasma Total Antioxidant ability (T-AOC) has been respectively increased 84.0%, 24.5% and 35.8%, 6h and 24h after infected by Aeromonas hydrophila fish body, Plasma MDA (MDA) content reduces by 17.2% and 3.1% respectively, 6h after infected by Aeromonas hydrophila fish body, what compare channel catfish key index level of plasma cortisol stress significantly increase 60.7% than before infecting, after drop sharply to about 18% before infection, and stablize constant, and according to the channel catfish after processing of step A after infected by Aeromonas hydrophila, level of plasma cortisol is not notable to rise, and drops to the level similar with matched group on the contrary after infecting 24h.
Relative to the channel catfish (comparison fish) that whole period of surviving the winter is not subject to any interference, do not have significant change according to the gene expression amount of heat stress proteins HSP70 and Ia interleukin I L, Toll-like receptor TLR and NOD in spleen when terminating, kidney and head-kidney of surviving the winter of the channel catfish after processing of step A.But, 24h after infected by Aeromonas hydrophila, the gene expression amount of the heat stress proteins HSP70 of comparison fish spleen and head-kidney than improves 278.5% and 128.2% before infecting respectively, then only than improves 58.5% and 54.0% before infecting respectively according to the gene expression amount of the channel catfish spleen after processing of step A and the heat stress proteins HSP70 of head-kidney; 48h after infected by Aeromonas hydrophila, improves 184.4% and 88.4% respectively according to the gene expression amount of the channel catfish spleen after processing of step A and the TLR3 of kidney than matched group channel catfish; 48h after infected by Aeromonas hydrophila, improves 19.6% and 83.1% respectively according to the gene expression amount of channel catfish spleen NOD1 and the NOD2 after processing of step A than matched group channel catfish; 72h after infected by Aeromonas hydrophila, improves 505.2% and 1325.7% respectively according to the gene expression amount of channel catfish spleen NOD1 and the NOD2 after processing of step A than matched group channel catfish; 72h after infected by Aeromonas hydrophila, improves 234.1% and 1739.0% respectively according to the gene expression amount of the interleukin I L ��-a of the channel catfish spleen after processing of step A and kidney than matched group channel catfish; 72h after infected by Aeromonas hydrophila, improves 305.0% and 515.3% respectively according to the gene expression amount of the interleukin I L ��-b of the channel catfish spleen after processing of step A and kidney than matched group channel catfish; 72h after infected by Aeromonas hydrophila, improves 165.4% and 645.3% respectively according to the gene expression amount of the channel catfish spleen after processing of step A and the interleukin I L8 of kidney than matched group channel catfish.
E, it is not subject to the Ctenopharyngodon idellus (comparison fish) of any interference relative to whole period of surviving the winter, after period cage culture Ctenopharyngodon idellus of surviving the winter being managed according to the aquaculture management method obtained in step A, the immunity of Ctenopharyngodon idellus of surviving the winter can be significantly increased, mainly include Ctenopharyngodon idellus survive the winter end time, significantly reducing lipid peroxidation product malonaldehyde (MDA) content 28.3% that can indirectly reflect cell injury in blood plasma, fish body stress key index level of plasma cortisol change little; 24h, 72h and 168h after infected by Aeromonas hydrophila fish body, blood plasma catalase (CAT) vigor has been respectively increased 47.7%, 101.9% and 15.8%; 6h after infected by Aeromonas hydrophila fish body, key index level of plasma cortisol stress reduce by 75%; 6h, 24h and 72h after infected by Aeromonas hydrophila fish body, Plasma MDA (MDA) content reduces by 34.3%, 48.0% and 23.1% respectively.
Relative to the Ctenopharyngodon idellus (comparison fish) that whole period of surviving the winter is not subject to any interference, do not have significant change according to the gene expression amount of heat stress proteins HSP70 and Ia Immunoglobulin IgM, Toll-like receptor TLR3 and major histocompatibility complex MHC in spleen when terminating, kidney and liver of surviving the winter of the Ctenopharyngodon idellus after processing of step A. But, 6h after infected by Aeromonas hydrophila, the gene expression amount of the heat stress proteins HSP70 of comparison Ctenopharyngodon idellus kidney and head-kidney than improves 2568.4% and 1660.6% before infecting respectively, then only than improves 504.6% and 731.2% before infecting respectively according to the gene expression amount of the Ctenopharyngodon idellus kidney after processing of step A and the heat stress proteins HSP70 of head-kidney; 6h after infected by Aeromonas hydrophila, improves 49.5% and 230.8% respectively according to the gene expression amount of the Ctenopharyngodon idellus kidney after processing of step A and spleen IgM than matched group Ctenopharyngodon idellus;24h after infected by Aeromonas hydrophila, improves 181.6% and 46.7% respectively according to the gene expression amount of the Ctenopharyngodon idellus kidney after processing of step A and spleen IgM than matched group Ctenopharyngodon idellus; 24h after infected by Aeromonas hydrophila, improves 546.6% and 38.9% respectively according to the gene expression amount of the Liver of Ctenopharyngodon Idellus after processing of step A and kidney MHCII-�� than matched group Ctenopharyngodon idellus; 24h after infected by Aeromonas hydrophila, improves 72.2% according to the gene expression amount of the Ctenopharyngodon idellus spleen MyD88 after processing of step A than matched group Ctenopharyngodon idellus; 72h after infected by Aeromonas hydrophila, improves 49.9% according to the gene expression amount of the Ctenopharyngodon idellus kidney MyD88 after processing of step A than matched group Ctenopharyngodon idellus; 24h, 72h and 168h after infected by Aeromonas hydrophila, improve 124.9%, 46.4% and 178.0% than matched group Ctenopharyngodon idellus respectively according to the gene expression amount of the Ctenopharyngodon idellus splenic T LR3 after processing of step A.
The aquaculture management method that F, step A obtain can strengthen the immunity of the cultured fishes such as Carassius auratus, channel catfish, Ctenopharyngodon idellus and to the resistivity after courses of infection.
Described cage culture is pond net box cultivation, river course cage culture or cages in reservoir cultivation.
It is therein any that described cultured fishes are specially Carassius auratus, channel catfish, Ctenopharyngodon idellus.
At present, the method improving cultured fishes immunity of surviving the winter targetedly mainly has the drug use etc. after surviving the winter. But not only existing Drug resistance due to chemicals, and breeding environment can be polluted by the accumulation that medicine is in water, destroying breeding ecological environment, thus forming vicious cycle.
The present invention compares with existing technology and has the advantage that
The aquaculture management method of this period of surviving the winter, method operation, low cost, drug residue free simple, easy; Stress key index heat stress proteins and cortisol levels by regulating, significantly improve adaptibility to response when fish body suffers the exogenous stimulations such as disease attack after surviving the winter, pattern recognition receptors, chemotactic factor and nonspecific immunity key enzyme can be activated, and then improve Fish immunity after surviving the winter and the resistance to disease. Its advantage is:
1. the present invention be survive the winter period aquaculture management method optimization, to use object technology require relatively low, have operational approach simply, do not use medicine, suitable in advantages such as different types of cage cultures.
2. the overwinter management method that the present invention relates to can pass through adjustment stress key index heat stress proteins and cortisol levels, significantly improve adaptibility to response when fish body suffers the exogenous stimulations such as disease attack after surviving the winter, pattern recognition receptors, chemotactic factor and nonspecific immunity key enzyme can be activated, and then improve Fish immunity after surviving the winter and the resistance to disease.
3. the aquaculture management method that the present invention relates to is tested by aquatic animal, it was shown that can improve survive the winter period cage-farming immunocompetence, it is possible to increase the survival rate after bacteria infection.
Detailed description of the invention
The following is and the specific embodiment of the present invention is illustrated.
Embodiment 1:
A kind of method that can improve survive the winter period cage culture Carassius aurutus gibelio immunity and disease resistance, the steps include:
A, test process group: winter test two process groups of design, matched group is whole period glitch-free cage culture Carassius aurutus gibelio of surviving the winter; And test group drags net cage once at the whole period 08:00 in the morning on every Mondays that survives the winter, after making the Carassius aurutus gibelio in net cage expose 5 minutes in atmosphere, again fish and net cage are put back in water, with this, Fish that survive the winter in net cage carried out weekly air exposure and fish for stimulation;Matched group has 3 tests parallel respectively with test process group, totally 6 mouthfuls of net cages. Whole surviving the winter period, net cage Fish are not thrown something and fed man-made feeds.
B, winter test: test in the Changjiang river therefore carry out in 6 mouthfuls of net cages (2m �� 2m �� 2m) of river course (Lao He the Changjiang river four, Shishou City, Hubei Province large Chinese carp seed farm). During test, recording water temperature every day, the water temperature of December is 4 DEG C to 10.5 DEG C, and the water temperature in January is 3.5 DEG C to 8.0 DEG C; The water temperature in February is 5.5 DEG C to 9.0 DEG C; The water temperature in March is 9.0 DEG C to 14.0 DEG C. Illumination condition is natural lighting, and water pH value is about 7.5.
C, before winter test, test fish carries out fortification in net cage system, and the enriched nutritive phase is one month, during this period, (09:00 and 15:00) commodity in use feedstuff (Shashi, Jingzhou City of Hubei Province Tong Wei feed corporation,Ltd, feedstuff number 108) feeding experiment fish twice daily. When on-test, all fish fasting one day, randomly select neat specification, 50 tail fishes (about 317g/ tail) of apparent health are put in every mouthful of net cage, totally 6 mouthfuls of net cages. During whole winter test, all of cultured fishes are not thrown something and fed artifical compound feed.
D, challenge test: Aeromonas hydrophila (Aeromonashydrophila) strain is applied to 30 �� of C of BHI flat board and cultivates 24h. Use PBS washing plate 3 times, collect bacterium solution and use the centrifugal 10min collection bacterium of centrifuge 3500g. Bacterial concentration regulates to 1.5 �� 108cfu/mL��
E, when winter test terminates, in every mouthful of net cage, remaining 40 tail fishes adopt to put back to after lumbar injection Aeromonas hydrophila liquid 0.5mL and continue in former net cage to raise. Add up the mortality rate of 168h continuously. Before challenge test, after challenge test after 6h, challenge test after 24h, challenge test after 72h and challenge test 168h separately sampled.
Result shows: after matched group and process group Carassius aurutus gibelio are survived the winter, body weight all has decline, but does not have significant change, and illustrates that this overwinter management method Carassius aurutus gibelio is survived the winter body weight does not affect. relative to the Carassius aurutus gibelio (comparison fish) that whole period of surviving the winter is not subject to any interference, after period cage culture Carassius aurutus gibelio of surviving the winter being managed according to the aquaculture management method obtained in step A, immunity and the viral resistance of Carassius aurutus gibelio of surviving the winter can be significantly increased, mainly include Carassius aurutus gibelio survive the winter end time, catalase (CAT) vigor 86% of fish body blood plasma can be significantly improved, improve Plasma Total Antioxidant ability (T-AOC) 36.5%, improve blood plasma total number born (T-SOD) vigor 14.5%, significantly reduce lipid peroxidation product malonaldehyde (MDA) content 21.9% that can indirectly reflect cell injury in blood plasma. these are it is shown that this overwinter management way can significantly improve the non-specific immunity of cage-farming when terminating of surviving the winter.
Described cultured fishes are that Carassius auratus, channel catfish, Ctenopharyngodon idellus are therein any. The immunity of cage-farming of surviving the winter can be obviously enhanced, effectively reduce the mortality rate after bacteria infection.
After infected by Aeromonas hydrophila fish body 168h, it is possible on the basis of comparison Carassius aurutus gibelio cumulative mortality 76.9%, reduce by 20.6% and reach the low actual of 56.3%; 6h, 24h and 72h after infected by Aeromonas hydrophila, blood plasma catalase (CAT) vigor has been respectively increased 181.5%, 56.8% and 37.1%; 6h and 24h after infected by Aeromonas hydrophila, Acute Coronary Syndrome (MPO) vigor has been respectively increased 33.3% and 117.6%;72h and 168h after infected by Aeromonas hydrophila, Plasma Total Antioxidant ability (T-AOC) has been respectively increased 51.9% and 59.1%; 72h after infected by Aeromonas hydrophila, Plasma MDA (MDA) content have dropped 40.6%; After infected by Aeromonas hydrophila, sharply stress falling after rise after the first notable rising of key index level of plasma cortisol of comparison Carassius aurutus gibelio, 6h after counteracting toxic substances reaches the highest (increasing 293.4%), and does not have significant change according to the Carassius aurutus gibelio cortisol levels after processing of step A after counteracting toxic substances. These, it is shown that this overwinter management way can significantly improve the cage-farming survival rate when suffering germ attack when surviving the winter end, also significantly improve at the metainfective nonspecific immunity key enzyme vigor of antibacterial.
Relative to the Carassius aurutus gibelio (comparison fish) that whole period of surviving the winter is not subject to any interference, according to the Carassius aurutus gibelio after processing of step A survive the winter heat stress proteins HSP70 and Ia interleukin I L-11 in spleen when terminating, kidney and head-kidney, marrow sample differentiation factor MyD88 and Toll-like receptor TLR3 gene expression amount do not have significant change. But, 6h after infected by Aeromonas hydrophila, the gene expression amount of the heat stress proteins HSP70 of comparison fish spleen, kidney and head-kidney than improves 242.2%, 145.8% and 16.9 times before infecting respectively, then only than improves 142.0%, 99.9% and 1.6 times before infecting respectively according to the gene expression amount of the heat stress proteins HSP70 of the Carassius aurutus gibelio spleen after processing of step A, kidney and head-kidney; 24h after infected by Aeromonas hydrophila, the gene expression amount of the interleukin I L-11 of comparison fish spleen, kidney and head-kidney than improves 2490.9 times, 57.8 times and 268.1 times before infecting respectively, then only than improves 1381.5 times, 33.2 times and 19.5 times before infecting respectively according to the gene expression amount of the interleukin I L-11 of the Carassius aurutus gibelio spleen after processing of step A, kidney and head-kidney; 6h after infected by Aeromonas hydrophila, the gene expression amount of the marrow sample differentiation factor MyD88 of comparison fish spleen, kidney and head-kidney than improves 86.8%, 87.7% and 229.8% before infecting respectively, then only improves 0.9%, 3.7% and 68.2% than before infecting respectively according to the gene expression amount of the marrow sample differentiation factor MyD88 of the Carassius aurutus gibelio spleen after processing of step A, kidney and head-kidney. These results show, this overwinter management way can significantly regulate stress the gene expression dose of key index heat stress proteins, raising fish body suffers the adaptibility to response during exogenous stimulations such as disease attack after surviving the winter, significantly improve the gene expression dose of pattern recognition receptors and chemotactic factor after courses of infection, and then improve Fish immunity after surviving the winter and the resistance to disease.
Embodiment 2:
A kind of method that can improve survive the winter period cage culture channel catfish immunity and disease resistance, the steps include:
A, test process group: winter test two process groups of design, matched group is whole period glitch-free cage culture channel catfish of surviving the winter; And test group drags net cage once at the whole period 08:00 in the morning on every Mondays that survives the winter, after making the channel catfish in net cage expose 4 minutes in atmosphere, again fish and net cage are put back in water, with this, Fish that survive the winter in net cage carried out weekly air exposure and fish for stimulation; Matched group has 3 tests parallel respectively with test process group, totally 6 mouthfuls of net cages. Whole surviving the winter period, net cage Fish are not thrown something and fed man-made feeds.
B, winter test: test in the Changjiang river therefore carry out in 6 mouthfuls of net cages (2m �� 2m �� 2m) of river course (Lao He the Changjiang river four, Shishou City, Hubei Province large Chinese carp seed farm).During test, recording water temperature every day, the water temperature of December is 4 DEG C to 10.5 DEG C, and the water temperature in January is 3.5 DEG C to 8.0 DEG C; The water temperature in February is 5.5 DEG C to 9.0 DEG C; The water temperature in March is 9.0 DEG C to 14.0 DEG C. Illumination condition is natural lighting, and water pH value is about 7.5.
C, before winter test, test fish carries out fortification in net cage system, and the enriched nutritive phase is one month, during this period, (09:00 and 15:00) commodity in use feedstuff (Shashi, Jingzhou City of Hubei Province Tong Wei feed corporation,Ltd, feedstuff number 108) feeding experiment fish twice daily. When on-test, all fish fasting one day, randomly select neat specification, 35 tail fishes (about 188g/ tail) of apparent health are put in every mouthful of net cage, totally 6 mouthfuls of net cages. During whole winter test, all of cultured fishes are not thrown something and fed artifical compound feed.
D, challenge test: Aeromonas hydrophila (Aeromonashydrophila) strain is applied to 30 �� of C of BHI flat board and cultivates 24h. Use PBS washing plate 3 times, collect bacterium solution and use the centrifugal 10min collection bacterium of centrifuge 3500g. Bacterial concentration regulates to 5.6 �� 108cfu/mL��
E, when winter test terminates, in every mouthful of net cage, remaining 30 tail fishes adopt to put back to after lumbar injection Aeromonas hydrophila liquid 0.5mL and continue in former net cage to raise. Before challenge test, after challenge test after 6h, challenge test after 24h, challenge test after 48h, challenge test after 72h and challenge test 168h separately sampled.
Result shows: after matched group and process group channel catfish are survived the winter, body weight does not have significant change, and illustrates that this overwinter management method channel catfish is survived the winter body weight does not affect. relative to the channel catfish (comparison fish) that whole period of surviving the winter is not subject to any interference, after period cage culture channel catfish of surviving the winter being managed according to the aquaculture management method obtained in step A, the immunity of channel catfish of surviving the winter can be significantly increased, mainly include channel catfish survive the winter end time, catalase (CAT) vigor 53.4% of fish body blood plasma can be significantly improved, significantly reduce lipid peroxidation product malonaldehyde (MDA) content 15.4% that can indirectly reflect cell injury in blood plasma, key index level of plasma cortisol stress significantly reduce 37.8%. these are it is shown that this overwinter management way can significantly improve the non-specific immunity of cage-farming when terminating of surviving the winter.
6h after infected by Aeromonas hydrophila fish body, 24h, 48h, 72h and 168h, blood plasma catalase (CAT) vigor has been respectively increased 64.7%, 65.6%, 79.3%, 48.9% and 27.4%; 24h, 48h and 72h after infected by Aeromonas hydrophila, Acute Coronary Syndrome (MPO) vigor has been respectively increased 39.8%, 13.3% and 32.5%; 6h, 24h and 48h after infected by Aeromonas hydrophila, Plasma Total Antioxidant ability (T-AOC) has been respectively increased 84.0%, 24.5% and 35.8%; 6h and 24h after infected by Aeromonas hydrophila fish body, Plasma MDA (MDA) content reduces by 17.2% and 3.1% respectively; 6h after infected by Aeromonas hydrophila fish body, what compare channel catfish key index level of plasma cortisol stress significantly increase 60.7% than before infecting, after drop sharply to about 18% before infection, and stablize constant, and according to the channel catfish after processing of step A after infected by Aeromonas hydrophila, level of plasma cortisol is not notable to rise, and drops to the level similar with matched group on the contrary after infecting 24h. These are it is shown that this overwinter management way can significantly improve the cage-farming nonspecific immunity key enzyme vigor when suffering germ attack of surviving the winter.
Relative to the channel catfish (comparison fish) that whole period of surviving the winter is not subject to any interference, do not have significant change according to the gene expression amount of heat stress proteins HSP70 and Ia interleukin I L, Toll-like receptor TLR and NOD in spleen when terminating, kidney and head-kidney of surviving the winter of the channel catfish after processing of step A.But, 24h after infected by Aeromonas hydrophila, the gene expression amount of the heat stress proteins HSP70 of comparison fish spleen and head-kidney than improves 278.5% and 128.2% before infecting respectively, then only than improves 58.5% and 54.0% before infecting respectively according to the gene expression amount of the channel catfish spleen after processing of step A and the heat stress proteins HSP70 of head-kidney; 48h after infected by Aeromonas hydrophila, improves 184.4% and 88.4% respectively according to the gene expression amount of the channel catfish spleen after processing of step A and the TLR3 of kidney than matched group channel catfish; 48h after infected by Aeromonas hydrophila, improves 19.6% and 83.1% respectively according to the gene expression amount of channel catfish spleen NOD1 and the NOD2 after processing of step A than matched group channel catfish; 72h after infected by Aeromonas hydrophila, improves 505.2% and 1325.7% respectively according to the gene expression amount of channel catfish spleen NOD1 and the NOD2 after processing of step A than matched group channel catfish; 72h after infected by Aeromonas hydrophila, improves 165.4% and 645.3% respectively according to the gene expression amount of the channel catfish spleen after processing of step A and the interleukin I L8 of kidney than matched group channel catfish; 72h after infected by Aeromonas hydrophila, improves 234.1% and 1739.0% respectively according to the gene expression amount of the interleukin I L ��-a of the channel catfish spleen after processing of step A and kidney than matched group channel catfish; 72h after infected by Aeromonas hydrophila, improves 305.0% and 515.3% respectively according to the gene expression amount of the interleukin I L ��-b of the channel catfish spleen after processing of step A and kidney than matched group channel catfish. These results show, this overwinter management way can significantly regulate stress the gene expression dose of key index heat stress proteins, raising fish body suffers the adaptibility to response during exogenous stimulations such as disease attack after surviving the winter, significantly improve the gene expression dose of pattern recognition receptors and chemotactic factor after courses of infection, and then improve Fish immunity after surviving the winter and the resistance to disease.
Embodiment 3:
A kind of method that can improve survive the winter period cage culture Ctenopharyngodon idellus immunity and disease resistance, the steps include:
A, test process group: winter test two process groups of design, matched group is whole period glitch-free cage culture Ctenopharyngodon idellus of surviving the winter; And test group drags net cage once at the whole period 08:00 in the morning on every Mondays that survives the winter, after making the Ctenopharyngodon idellus in net cage expose 6 minutes in atmosphere, again fish and net cage are put back in water, with this, Fish that survive the winter in net cage carried out weekly air exposure and fish for stimulation; Matched group has 3 tests parallel respectively with test process group, totally 6 mouthfuls of net cages. Whole surviving the winter period, net cage Fish are not thrown something and fed man-made feeds.
B, winter test: test in the Changjiang river therefore carry out in 6 mouthfuls of net cages (2m �� 2m �� 2m) of river course (Lao He the Changjiang river four, Shishou City, Hubei Province large Chinese carp seed farm). During test, recording water temperature every day, the water temperature of December is 4 DEG C to 10.5 DEG C, and the water temperature in January is 3.5 DEG C to 8.0 DEG C; The water temperature in February is 5.5 DEG C to 9.0 DEG C; The water temperature in March is 9.0 DEG C to 14.0 DEG C. Illumination condition is natural lighting, and water pH value is about 7.5.
C, before winter test, test fish carries out fortification in net cage system, and the enriched nutritive phase is one month, during this period, (09:00 and 15:00) commodity in use feedstuff (Shashi, Jingzhou City of Hubei Province Tong Wei feed corporation,Ltd, feedstuff number 108) feeding experiment fish twice daily.When on-test, all fish fasting one day, randomly select neat specification, 25 tail fishes (about 778g/ tail) of apparent health are put in every mouthful of net cage, totally 6 mouthfuls of net cages. During whole winter test, all of cultured fishes are not thrown something and fed artifical compound feed.
D, challenge test: Aeromonas hydrophila (Aeromonashydrophila) strain is applied to 30 �� of C of BHI flat board and cultivates 24h. Use PBS washing plate 3 times, collect bacterium solution and use the centrifugal 10min collection bacterium of centrifuge 3500g. Bacterial concentration regulates to 6.8 �� 108cfu/mL��
E, when winter test terminates, in every mouthful of net cage, remaining 22 tail fishes adopt to put back to after lumbar injection Aeromonas hydrophila liquid 0.5mL and continue in former net cage to raise. Before challenge test, after challenge test after 6h, challenge test after 24h, challenge test after 72h and challenge test 168h separately sampled.
Result shows: after matched group and process group Ctenopharyngodon idellus are survived the winter, body weight does not have significant change, and illustrates that this overwinter management method Ctenopharyngodon idellus is survived the winter body weight does not affect. Relative to the Ctenopharyngodon idellus (comparison fish) that whole period of surviving the winter is not subject to any interference, after period cage culture Ctenopharyngodon idellus of surviving the winter being managed according to the aquaculture management method obtained in step A, the immunity of Ctenopharyngodon idellus of surviving the winter can be significantly increased, mainly include Ctenopharyngodon idellus survive the winter end time, significantly reducing lipid peroxidation product malonaldehyde (MDA) content 28.3% that can indirectly reflect cell injury in blood plasma, fish body stress key index level of plasma cortisol change little; 24h, 72h and 168h after infected by Aeromonas hydrophila fish body, blood plasma catalase (CAT) vigor has been respectively increased 47.7%, 101.9% and 15.8%; 6h after infected by Aeromonas hydrophila fish body, key index level of plasma cortisol stress reduce by 75%; 6h, 24h and 72h after infected by Aeromonas hydrophila fish body, Plasma MDA (MDA) content reduces by 34.3%, 48.0% and 23.1% respectively. These are it is shown that this overwinter management way can significantly improve the cage-farming non-specific immunity when attacked by pathogenic bacteria of surviving the winter.
Relative to the Ctenopharyngodon idellus (comparison fish) that whole period of surviving the winter is not subject to any interference, do not have significant change according to the gene expression amount of heat stress proteins HSP70 and Ia Immunoglobulin IgM, Toll-like receptor TLR3 and major histocompatibility complex MHC in spleen when terminating, kidney and liver of surviving the winter of the Ctenopharyngodon idellus after processing of step A. But, 6h after infected by Aeromonas hydrophila, the gene expression amount of the heat stress proteins HSP70 of comparison Ctenopharyngodon idellus kidney and head-kidney than improves 2568.4% and 1660.6% before infecting respectively, then only than improves 504.6% and 731.2% before infecting respectively according to the gene expression amount of the Ctenopharyngodon idellus kidney after processing of step A and the heat stress proteins HSP70 of head-kidney; 6h after infected by Aeromonas hydrophila, improves 49.5% and 230.8% respectively according to the gene expression amount of the Ctenopharyngodon idellus kidney after processing of step A and spleen IgM than matched group Ctenopharyngodon idellus; 24h after infected by Aeromonas hydrophila, improves 181.6% and 46.7% respectively according to the gene expression amount of the Ctenopharyngodon idellus kidney after processing of step A and spleen IgM than matched group Ctenopharyngodon idellus; 24h after infected by Aeromonas hydrophila, improves 546.6% and 38.9% respectively according to the gene expression amount of the Liver of Ctenopharyngodon Idellus after processing of step A and kidney MHCII-�� than matched group Ctenopharyngodon idellus; 24h after infected by Aeromonas hydrophila, improves 72.2% according to the gene expression amount of the Ctenopharyngodon idellus spleen MyD88 after processing of step A than matched group Ctenopharyngodon idellus;72h after infected by Aeromonas hydrophila, improves 49.9% according to the gene expression amount of the Ctenopharyngodon idellus kidney MyD88 after processing of step A than matched group Ctenopharyngodon idellus; 24h, 72h and 168h after infected by Aeromonas hydrophila, improve 124.9%, 46.4% and 178.0% than matched group Ctenopharyngodon idellus respectively according to the gene expression amount of the Ctenopharyngodon idellus splenic T LR3 after processing of step A. These results show, this overwinter management way can significantly regulate stress the gene expression dose of key index heat stress proteins, raising fish body suffers the adaptibility to response during exogenous stimulations such as disease attack after surviving the winter, by significantly improving the gene expression dose of pattern recognition receptors after courses of infection, chemotactic factor and immunoglobulin, and then improve cage-farming immunity after surviving the winter and the resistance to disease.
Also, it should be noted those skilled in the art can also make such or such easy variation pattern, such as equivalent way, or obvious mode of texturing under the instruction of this specification; Above-mentioned variation pattern all should within protection scope of the present invention.
Claims (2)
1. raising is survived the winter period cage culture Carassius aurutus gibelio immunity and the method for disease resistance, the steps include:
A, test process group: winter test one test group of design and a matched group, matched group is whole period glitch-free cage culture Carassius aurutus gibelio of surviving the winter, test group drags net cage once for the period 08:00 in the morning on every Mondays that survives the winter, after making the Carassius aurutus gibelio in net cage expose 4-6 minute in atmosphere, again fish and net cage are put back in water, the Fish that survive the winter in net cage carrying out weekly air exposure and fishes for stimulation, whole surviving the winter period, net cage Fish are not thrown something and fed man-made feeds;
B, winter test: carry out in 6 mouthfuls of net cage: 2m �� 2m �� 2m in event river course, the Changjiang river, test group and matched group 3 mouthfuls of net cages of random assortment respectively, during test, every day records water temperature, the water temperature of December is 4 DEG C to 10.5 DEG C, and the water temperature in January is 3.5 DEG C to 8.0 DEG C; The water temperature in February is 5.5 DEG C to 9.0 DEG C; The water temperature in March is 9.0 DEG C to 14.0 DEG C, and illumination condition is natural lighting, and water pH value is 7.3-7.5;
C, before winter test, test fish carries out fortification in net cage system, the enriched nutritive phase is one month, during this period, twice daily: 09:00 and 15:00 commodity in use feedstuff feeding experiment fish, when on-test, all fish fasting one day, randomly select neat specification, and the fish of apparent health is put in every mouthful of net cage, during whole winter test, all of cultured fishes are not thrown something and fed artifical compound feed;
D, challenge test: Aeromonas hydrophila strain is applied to 30 �� of C of BHI flat board and cultivates 24h, uses PBS washing plate 3 times, collect bacterium solution and use the centrifugal 10min collection bacterium of centrifuge 3500g, and bacterial concentration regulates to 1.5 �� 108Cfu/mL;
E, winter test terminate, in every mouthful of net cage, remaining fish is put back to after adopting lumbar injection Aeromonas hydrophila liquid 0.5mL in former net cage and continues to raise, add up continuously the mortality rate of 168h, before challenge test, after challenge test after 6h, challenge test after 24h, challenge test after 72h and challenge test 168h separately sampled.
2. a kind of raising according to claim 1 survives the winter period cage culture Carassius aurutus gibelio immunity and the method for disease resistance, described Carassius aurutus gibelio channel catfish or Ctenopharyngodon idellus replace.
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