A method for in-situ treatment of bottom sediment in cage culture using bacterial flora-Echinops monocyclicus combined

Abstract

The present invention relates to the technical field of marine environmental protection. The present invention provides a method for in-situ treatment of cage culture bottom using a bacterial conglomerate and a monocyprid, comprising the following steps: (1) conducting stress-resistant and enhanced culture on monocyprid seedlings to obtain enhanced monocyprid seedlings; (2) introducing the enhanced monocyprid seedlings and bacterial conglomerates into a culture area; (3) managing the monocyprid seedlings after introduction. The present invention adopts a pollution treatment method using a bacterial conglomerate and a monocyprid based on the nutritional characteristics of the bottom sludge of offshore aquaculture, and in particular adopts an efficient implantation method to introduce the monocyprid and bacterial conglomerates, so that an ecosystem is formed at the bottom of the aquaculture cage area. The system can be applied to fish leftover bait and metabolic waste, so as to achieve the purpose of purifying the aquaculture bottom, protecting the offshore ecological environment, and improving economic benefits.

Description

Technical Field

[0001] The invention relates to the technical field of marine environmental protection, and in particular to an in-situ treatment method for a bacterial colony-Echinococcus monocynoides-combined cage aquaculture bottom sediment. Background Art

[0002] In recent years, with concerns about a series of social and ecological problems caused by land-based aquaculture, such as land occupation, high energy consumption, frequent diseases, product quality decline, and excessive discharge of aquaculture wastewater, the call for shifting from land to offshore and deep-sea aquaculture has become increasingly louder. Therefore, a new modern aquaculture model that focuses on offshore and offshore aquaculture and is supplemented by land-based aquaculture has gradually emerged, taking into account social, ecological, and economic benefits.

[0003] As a necessary link between land-based seedling cultivation and deep-sea adult fish farming, offshore aquaculture determines the final aquaculture output and benefits. Offshore aquaculture is to preserve young fish and strengthen fattening of fish in the sea area close to the coast to adapt to the harsh environment of deep-sea aquaculture and improve the success rate of aquaculture. In offshore aquaculture, in order to make it grow rapidly, fresh fish, low-value fish and artificial feed are often used excessively for intensive feeding. Due to the limited digestive capacity of fish and the large amount of feces produced, some indigestible residual bait and feces will settle to the bottom of the aquaculture cages and accumulate, becoming the main source of pollution. Since the aquaculture location is close to the coast or in the bay, and a large number of cages form a shielding effect, the ocean current cannot take away pollutants in time, resulting in the deterioration of the environment at the bottom of the aquaculture cages, and the sludge accumulates more and more, providing conditions for the breeding and reproduction of pathogens, resulting in frequent diseases, the death of a large number of aquatic animals, and the worsening of the aquaculture environment, forming a secondary pollution source.

[0004] The accumulation rate of organic pollution in aquaculture sediments exceeds the decomposition capacity, which is the root cause of environmental degradation. How to utilize these excrement and leftover bait from the source and improve the pollution self-purification capacity of the aquaculture environment is an urgent problem to be solved. Some current restoration technologies are limited to small water bodies such as aquaculture ponds, and the treatment of nearshore cage aquaculture sediments is basically a technical gap. The treatment methods for pond sediments are relatively simple, usually drying sludge, using lime and other improvers, removing sediments, or using biological control methods, etc. However, these methods cannot be carried out in open or semi-open seas.

[0005] The fundamental solution to this problem lies in the invention of a simple, effective, widely applicable and economically beneficial in-situ bioremediation technology for sediments. The monocyclic aphid is a burrowing organism. Its special behavior of disturbing the sediment is more effective in improving the sediment environment. Studies have found that the sulfide content in cage aquaculture sediments is relatively high. The monocyclic aphid is regarded as a model organism for sulfide metabolism research. It has a high tolerance, oxidative detoxification and metabolic adaptation to sulfides. Therefore, as a large organism, its ability to handle sulfides is more efficient than that of microorganisms. Bacterial flora, as a carrier that can accommodate a variety of symbiotic bacteria, can not only improve the survival rate of bacteria, provide a "base" for the reproduction and spread of bacteria, but also provide a transitional medium for bacteria to enter the aquaculture sediment. Summary of the invention

[0006] The purpose of the present invention is to provide a method for in-situ treatment of cage aquaculture bottom sediment using a bacterial flora-Spinipes monocyclica combination, by introducing large organisms and Bacillus subtilis and the like to the bottom of offshore aquaculture cages to establish a local ecological nutrition chain of bacteria-Spinipes monocyclica-cage cultured fish, so that fish leftover bait and feces are ingested and utilized by Spinipes monocyclica in the process of entering the bottom sediment, and finally the metabolites are utilized by Bacillus subtilis and indigenous bacteria, thereby achieving the effects of effective treatment of aquaculture bottom sediment at the source stage, increased economic benefits of aquaculture, and improved self-purification capacity of aquaculture ecological environment.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides a method for in-situ treatment of bottom sediment in cage culture by using bacterial flora-Echinops unicinctus, comprising the following steps:

[0009] (1) performing stress-resistant strengthening cultivation on the seedlings of the single-ringed worm to obtain strengthened seedlings of the single-ringed worm;

[0010] (2) introducing the enhanced monocyclic worm seedlings and bacterial colonies into the culture area;

[0011] (3) Carry out management after the introduction of single-ringed spiny moth seedlings.

[0012] Preferably, the weight of the seedlings of the monocyclic moth in step (1) is 0.3-1.0 g and the body length is 20-50 mm.

[0013] Preferably, the stress-resistant enhancement culture in step (1) includes feed stress-resistant enhancement and environmental stress-resistant enhancement;

[0014] The method of strengthening the stress resistance of the bait is: before the monocyclic worm seedlings are introduced into the breeding area, the stress-strengthening bait is continuously fed for 5 to 14 days, and the amount of the stress-strengthening bait is 1 to 2% of the body weight of the monocyclic worm seedlings per day;

[0015] The stress-resistant and strengthened bait comprises the following raw materials in parts by weight: 25-45 parts of ground seaweed liquid, 3-15 parts of fish meal, 3-10 parts of starch, 5-20 parts of ultrafinely crushed fermented soybean meal, 25-45 parts of sea mud, 0.01-0.05 parts of yeast selenium ultrafine powder, 4-12 parts of scallop edge waste powder, 1-3 parts of sodium alginate, 0.5-2.0 parts of vitamin premix, 0.5-1.5 parts of mineral premix, 0.1-0.5 parts of taurine magnesium, 0.02-0.1 parts of tea polyphenols, and 0.04-0.15 parts of fermented all-trans vitamin K;

[0016] The preparation method of the stress-resistant and strengthened bait is:

[0017] A. Grinding fish meal, starch, ultrafinely pulverized fermented soybean meal, ultrafine yeast selenium powder and scallop edge waste powder into 150-200 meshes to obtain a mixed powder;

[0018] B. mixing the mixed powder, ground seaweed liquid, sea mud, sodium alginate, vitamin premix, mineral premix, magnesium taurine, tea polyphenols and fermented all-trans vitamin K to obtain a mixture;

[0019] C. Add water to the mixture and stir, and extrude it into granules with a diameter of 1 to 4 mm and a height of 1 to 3 mm;

[0020] D. drying the pellets to a moisture content of less than 8%, and then crushing them into fine particles with a particle size of 50 to 100 meshes to obtain the stress-resistant and enhanced bait;

[0021] The amount of water added in step C is 10-30% of the mass of the mixed material. The pellet extruder is used for extrusion, and the extrusion density during extrusion is set to 1.2-1.5 g / cm 3 .

[0022] Environmental stress resistance strengthening: mainly for hypoxia resistance strengthening, that is, by adjusting the aeration volume, the dissolved oxygen level is slowly reduced from 7 to 8 mg / L to 1.5 to 2 mg / L at a rate of 0.5 to 1 mg / L per day. Environmental stress resistance strengthening is carried out simultaneously with bait stress resistance strengthening, that is, the dissolved oxygen level is slowly reduced while bait stress resistance strengthening is being carried out. Environmental stress resistance strengthening lasts for 10 to 14 days, and at the end of stress resistance strengthening, the lowest oxygen level is reduced to 1.5 to 2 mg / L.

[0023] Preferably, the method for introducing the reinforced monocyclic schizont seedlings in step (2) is as follows: the reinforced monocyclic schizont seedlings are placed in the offshore waters at a distance of 0 to 20 cm from the seabed, a seedling placing device is used for the introduction, the temperature during the introduction is 16 to 25° C., the sea current speed is 0 to 0.1 m / s, and the sea state is 0 to 2 levels; the density of the introduced monocyclic schizont seedlings is 30 to 70 per m 2 .

[0024] Preferably, the method for introducing the bacterial mass in step (2) is: introducing the bacterial mass at a concentration of 150 to 300 g / m 2 The standard is to spray it in the breeding area.

[0025] Preferably, the method for preparing the bacterial cluster in step (2) comprises the following steps:

[0026] ① After crushing the volcanic rocks, soak them in seawater for 3 to 7 days, then add nutrient salts and continue soaking for 1 to 2 days to obtain soaked volcanic rocks;

[0027] ② respectively expanding and culturing the nitrate-reducing Marinobacter, the chemoautotrophic sulfur-oxidizing bacteria, the red bacillus and the salt sediment slow-growing Bacillus to obtain the nitrate-reducing Marinobacter bacterial solution, the chemoautotrophic sulfur-oxidizing bacteria bacterial solution, the red bacillus bacterial solution and the salt sediment slow-growing Bacillus bacterial solution;

[0028] ③ Add the nitrate-reducing Marinobacterium bacterial solution, the chemoautotrophic sulfur-oxidizing bacteria bacterial solution, the red bacillus bacterial solution and the salt sediment slow-growing Bacillus bacterial solution into the seawater of step ①, continue to soak, and obtain bacterial flora.

[0029] Preferably, the particle size of the crushed volcanic rock in step ① is 0.2-0.8 mm, and the mass ratio of the volcanic rock to seawater is 1:1-3;

[0030] The seawater after adding nutrient salts includes the following raw materials by weight: 3-5% (NH 4 ) 2 SO 4 , 0.5~0.85%KNO 3 1.0~1.6%CaCl 2 ·2H 2 O, 2.0-3.0% MgSO 4 7H 2 O, 2.0~4.0%K 2 HPO 4 1.8~3.8%KH 2 PO 4 , 0.1~0.4%FeSO 4 7H 2 O, 0.2~0.4%EDTA-Na 2 , 0.005~0.02%NaMoO 4 ·2H 2 O, 0.01~0.03% MnCl 2 , 0.00005~0.00035%CoCl 2 6H 2 O, 0.005~0.025%ZnSO 4 7H2 O, 0.0015~0.0025%CuSO 4 ·5H 2 O, the remainder is sea water;

[0031] The method for expanding the culture in step ② is: inoculate the bacteria into 2216E liquid culture medium at a rate of 1-5%, and culture until 10 4 ~10 6 Pieces / ml;

[0032] The volume ratio of the nitrate-reducing Marinobacterium solution, the chemoautotrophic sulfur-oxidizing bacteria solution, the Erythrobacterium solution, the salt sediment bradybacillus solution and the seawater in step ③ is 1-2:1-2:1-2:1-2:1000;

[0033] The temperature of the continued soaking is 20-28° C., and the time of the continued soaking is 5-14 days.

[0034] Preferably, the post-introduction management method in step (3) is: feeding cage fish feed on the day of introduction, the 2nd, 4th to 5th, 7th to 8th, 10th to 12th, and 15th to 17th day; selecting 2 to 4 1m2 cages on the day of introduction, the 15th, 30th, 90th, and 150th day, respectively; 2 Excavate the seabed area, calculate the survival rate of Echinops monocyrthistus on the 15th, 30th, 90th and 150th days, and carry out 1 to 3 replenishment according to the survival rate;

[0035] The calculation formula of the retention rate is:

[0036]

[0037] When the retention rate is lower than 50%, supplementary seedlings are carried out.

[0038] The present invention provides a method for in-situ treatment of cage culture bottom using a bacterial flora-Echinops monocylindrica combination, comprising the following steps: (1) performing stress-resistant and enhanced culture on Echinops monocylindrica seedlings to obtain enhanced Echinops monocylindrica seedlings; (2) introducing the enhanced Echinops monocylindrica seedlings and bacterial flora into a culture area; (3) managing the Echinops monocylindrica seedlings after introduction. The present invention adopts a pollution treatment method using a bacterial flora-Echinops monocylindrica combination based on the nutritional characteristics of the bottom mud of offshore aquaculture, and in particular, adopts an efficient implantation method to introduce Echinops monocylindrica and bacterial flora, so that an ecosystem is formed at the bottom of the aquaculture cage area. The system can be used for fish residual bait and metabolic waste, so as to achieve the purpose of purifying the aquaculture bottom, protecting the ecological environment of offshore waters, and improving economic benefits. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1This is the appearance of the seedling release device used when introducing the monocyclic worm;

[0040] Figure 2 The sulfide content in the bottom mud of each experimental group area 5 months after the introduction of Echinops monocyclica in Example 1;

[0041] Figure 3 The total carbon content in the sediment of each experimental group in Example 1 at 5 months after the introduction of Echinops monocyclica;

[0042] Figure 4 The total phosphorus content in the sediment of each experimental group in Example 1 5 months after the introduction of Echinops monocyclica;

[0043] Figure 5 The total nitrogen content in the bottom mud of each experimental group in Example 1 at 5 months after the introduction of Echinops monocyclica;

[0044] Figure 6 The sulfide content in the bottom mud of each experimental sea area 5 months after the introduction of Echinops monocyclica in Example 2;

[0045] Figure 7 The total carbon content in the bottom mud of each experimental sea area 5 months after the introduction of Echinops monocyclica in Example 2;

[0046] Figure 8 The total phosphorus content in the bottom mud of each experimental sea area 5 months after the introduction of Echinops monocyclica in Example 2;

[0047] Fig. 9 The total nitrogen content in the bottom mud of each experimental sea area 5 months after the introduction of Echinops monocyclica in Example 2; DETAILED DESCRIPTION

[0048] The present invention provides a method for in-situ treatment of bottom sediment in cage culture by using bacterial flora-Echinops unicinctus, comprising the following steps:

[0049] (1) performing stress-resistant strengthening cultivation on the seedlings of the single-ringed worm to obtain strengthened seedlings of the single-ringed worm;

[0050] (2) introducing the enhanced monocyclic worm seedlings and bacterial colonies into the culture area;

[0051] (3) Carry out management after the introduction of single-ringed spiny moth seedlings.

[0052] In the present invention, the weight of the monocyclic spiny worm seedlings in step (1) is preferably 0.3-1.0 g, more preferably 0.5-0.7 g, and the body length is preferably 20-50 mm, more preferably 30-40 mm.

[0053] In the present invention, the stress-resistant enhancement culture in step (1) preferably includes feed stress-resistant enhancement and environmental stress-resistant enhancement;

[0054] The method of strengthening the stress resistance of the feed is preferably: before the monocyclic spiny worm seedlings are introduced into the breeding area, the stress-strengthening feed is continuously fed for 5 to 14 days, and the amount of the stress-strengthening feed is 1 to 2% of the body weight of the monocyclic spiny worm seedlings / day. It is further preferably: before the monocyclic spiny worm seedlings are introduced into the breeding area, the stress-strengthening feed is continuously fed for 7 to 10 days, and the amount of the stress-strengthening feed is 1.5% of the body weight of the monocyclic spiny worm seedlings / day;

[0055] The stress-resistant and strengthened bait preferably comprises the following raw materials in parts by weight: 25-45 parts of ground seaweed liquid, 3-15 parts of fish meal, 3-10 parts of starch, 5-20 parts of ultrafinely crushed fermented soybean meal, 25-45 parts of sea mud, 0.01-0.05 parts of yeast selenium ultrafine powder, 4-12 parts of scallop edge waste powder, 1-3 parts of sodium alginate, 0.5-2.0 parts of vitamin premix, 0.5-1.5 parts of mineral premix, 0.1-0.5 parts of taurine magnesium, 0.02-0.1 parts of tea polyphenols, and fermented all-trans vitamin K. 0.04-0.15 parts, more preferably: 35 parts of ground seaweed liquid, 10 parts of fish meal, 7 parts of starch, 15 parts of ultrafinely crushed fermented soybean meal, 35 parts of sea mud, 0.03 parts of yeast selenium ultrafine powder, 8 parts of scallop edge waste powder, 2 parts of sodium alginate, 1.5 parts of vitamin premix, 1.0 parts of mineral premix, 0.3 parts of taurine magnesium, 0.06 parts of tea polyphenols, and 0.1 parts of fermented all-trans vitamin K;

[0056] In the present invention, the preparation method of the ground seaweed liquid is as follows: fresh kelp, enteromorpha and wakame within 1 to 3 days are collected, mixed in a mass ratio of 1 to 2:1 to 2:1 to 2, and ground.

[0057] In the present invention, the fish meal has a protein content of more than 45% and a fat content of less than 12%, and is purchased from Rongcheng New Hope Fish Meal Co., Ltd.

[0058] In the present invention, the starch has a purity of more than 95% and is purchased from Hebei Guangyu Starch Sugar Co., Ltd.

[0059] In the present invention, the ultrafinely pulverized fermented soybean meal has a particle size of more than 150 meshes, a protein content of more than 44%, and a fat content of less than 1%, and is purchased from Huamao (Shandong) Biotechnology Co., Ltd.

[0060] In the present invention, the sea mud is crushed to a particle size of 150 mesh or more and is purchased from Weihai Zhicheng Feed Co., Ltd.

[0061] In the present invention, the yeast selenium is crushed to a particle size of 150 mesh or more and is purchased from Shandong Pingju Biotechnology Co., Ltd.

[0062] In the present invention, the scallop edge waste is the scallop part removed from the scallop column, which is dried and crushed, and passed through a 150 mesh sieve.

[0063] In the present invention, the sodium alginate has a purity of more than 80% and is purchased from Henan Minghe Biotechnology Co., Ltd.

[0064] In the present invention, the vitamin premix composition is preferably (per kg feed): vitamin A 6600.00-8600.00 IU, vitamin D 1000.00-1400.00 IU, vitamin E 100.00-140.00 mg, vitamin K3 16.00-20.00 mg, vitamin B1 12.00-16.00 mg, vitamin B2 12.00-14.00 mg, vitamin B12 8.00-12.00 mg, folic acid 4.10-4.50 mg, choline 15.00-19.00 mg, pantothenic acid 55.00-65.00 mg, nicotinamide 37.00-41.00 mg, pyridoxine 65.00-75.00 mg, inositol 240.00-280.00 mg, vitamin C 300.00~320.00mg, vitamin H75.00~85.00mg, tert-butylparabenzoic acid 180~220mg, more preferably: vitamin A 7600.00IU, vitamin D 1200.00IU, vitamin E 120.00mg, vitamin K318.00mg, vitamin B114.00 mg, vitamin B213.00mg, vitamin B1210.00 mg, folic acid 4.30mg, choline 17.00mg, pantothenic acid 60.00mg, niacinamide 39.00mg, pyridoxine 70.00mg, inositol 260.00mg, vitamin C 310.00mg, vitamin H 80.00mg, tert-butylparabenzoic acid 200mg, each ingredient is purchased from Shandong Liuhe Group Co., Ltd.

[0065] In the present invention, the mineral premix composition is preferably (per kg feed): 55.00-65.00 mg manganese sulfate, 45.00-55.00 mg zinc sulfate, 35.00-45.00 mg iron sulfate, 18.00-22.00 mg copper methionine, 0.50-0.70 mg yeast selenium, 1.20-1.40 mg cobalt chloride, 2.50-3.50 mg potassium sulfate, and further preferably: 60.00 mg manganese sulfate, 50.00 mg zinc sulfate, 40.00 mg iron sulfate, 20.00 mg copper methionine, 0.60 mg yeast selenium, 1.30 mg cobalt chloride, 3.00 mg potassium sulfate, and each component is purchased from Shandong Liuhe Group Co., Ltd.

[0066] In the present invention, the magnesium taurine has a purity of 99% and is purchased from Ningbo Yingqian Technology Co., Ltd.

[0067] In the present invention, the tea polyphenols are purchased from Qingdao Chengrunda Food Ingredients Co., Ltd.

[0068] In the present invention, the fermented all-trans vitamin K has a purity of more than 0.2% and is purchased from Xi'an Tianbao Biotechnology Co., Ltd.

[0069] In the present invention, the preparation method of the stress-resistant and strengthened bait is:

[0070] A. Grinding fish meal, starch, ultrafinely pulverized fermented soybean meal, ultrafine yeast selenium powder and scallop edge waste powder into 150-200 meshes to obtain a mixed powder;

[0071] B. mixing the mixed powder, ground seaweed liquid, sea mud, sodium alginate, vitamin premix, mineral premix, magnesium taurine, tea polyphenols and fermented all-trans vitamin K to obtain a mixture;

[0072] C. Add water to the mixture and stir, and extrude it into granules with a diameter of 1 to 4 mm and a height of 1 to 3 mm;

[0073] D. drying the pellets to a moisture content of less than 8%, and then crushing them into fine particles with a particle size of 50 to 100 meshes to obtain the stress-resistant and strengthened bait;

[0074] In the present invention, the mesh size in step A is preferably 175 meshes.

[0075] In the present invention, the diameter in step C is preferably 2-3 mm, and the height is preferably 2 mm.

[0076] In the present invention, the particle size in step D is preferably 80 mesh.

[0077] In the present invention, the amount of water added in step C is preferably 10-30% of the mass of the mixed material, and more preferably 20%. The extrusion is preferably performed using a pellet extruder, and the extrusion density during the extrusion is preferably set to 1.2-1.5 g / cm 3 , more preferably 1.3 to 1.4 g / cm 3 .

[0078] In the present invention, the method of environmental stress strengthening is preferably: by adjusting the aeration volume, the dissolved oxygen is slowly reduced from 7 to 8 mg / L to 1.5 to 2 mg / L at a rate of 0.5 to 1 mg / L per day, and more preferably: by adjusting the aeration volume, the dissolved oxygen is slowly reduced from 7.5 mg / L to 1.7 to 1.8 mg / L at a rate of 0.7 to 0.8 mg / L per day. The environmental stress strengthening is preferably carried out simultaneously with the bait stress strengthening, and the duration of the environmental stress strengthening is preferably 10 to 14 days, and more preferably 12 days.

[0079] In the present invention, the method for introducing the reinforced monocyclic spiny worm seedlings in step (2) is preferably: placing the reinforced monocyclic spiny worm seedlings into the offshore waters at a distance of 0 to 20 cm from the seabed, more preferably at a distance of 10 cm from the seabed; preferably using a seedling releaser during the introduction; preferably the temperature during the introduction is 16 to 25°C, the current speed is 0 to 0.1 m / s, and the sea state is 0 to 2 levels, more preferably 20°C, the current speed is 0.05 m / s, and the sea state is 1 level; the density of the introduced monocyclic spiny worm seedlings is preferably 30 to 70 per m 2 , more preferably 50 pieces / m 2 .

[0080] In the present invention, the method for introducing the bacterial mass in step (2) is preferably: introducing the bacterial mass at a concentration of 150 to 300 g / m 2 The bacterial mass is sprayed in the breeding area according to the standard of 200-250g / m 2 The standard is to spray it in the breeding area.

[0081] In the present invention, the method for preparing the bacterial cluster in step (2) preferably comprises the following steps:

[0082] ① After crushing the volcanic rock, soak it in seawater for 3 to 7 days, then add nutrient salt and continue soaking for 1 to 2 days to obtain the soaked volcanic rock;

[0083] ② respectively expanding and culturing the nitrate-reducing Marinobacter, the chemoautotrophic sulfur-oxidizing bacteria, the red bacillus and the salt sediment slow-growing Bacillus to obtain the nitrate-reducing Marinobacter bacterial solution, the chemoautotrophic sulfur-oxidizing bacteria bacterial solution, the red bacillus bacterial solution and the salt sediment slow-growing Bacillus bacterial solution;

[0084] ③ Add the nitrate-reducing Marinobacterium bacterial solution, the chemoautotrophic sulfur-oxidizing bacteria bacterial solution, the red bacillus bacterial solution and the salt sediment slow-growing Bacillus bacterial solution into the seawater of step ①, continue to soak, and obtain bacterial flora.

[0085] In the present invention, the nitrate-reducing Marinobacterium (Marinobacterium nitratireducens) was purchased from Shanghai Xuanke Biotechnology Co., Ltd., the chemoautotrophic sulfur-oxidizing bacteria (Thiobacillus denitrificans) was purchased from Beijing Biobowei Biotechnology Co., Ltd., Erythrobacter flavus was purchased from Ningbo Mingzhou Biotechnology Co., Ltd., and the salt sediment slow-growing Bacillus (Lentibacillus sp) was purchased from Shanghai Xige Biotechnology Co., Ltd.

[0086] In the present invention, the particle size of the crushed volcanic rock in step ① is preferably 0.2-0.8 mm, more preferably 0.4-0.6 mm, the mass ratio of the volcanic rock to seawater is preferably 1:1-3, more preferably 1:2, and the seawater immersion time is preferably 3-7 days, more preferably 5 days;

[0087] The seawater after adding nutrient salts includes the following raw materials by weight: 3-5% (NH 4 ) 2 SO 4 , 0.5~0.85%KNO 3 1.0~1.6%CaCl 2 ·2H 2 O, 2.0-3.0% MgSO 4 7H 2 O, 2.0~4.0%K 2 HPO 4 1.8~3.8%KH 2 PO 4 , 0.1~0.4%FeSO 4 7H 2 O, 0.2~0.4%EDTA-Na 2 , 0.005~0.02%NaMoO 4 ·2H 2 O, 0.01~0.03% MnCl 2 , 0.00005~0.00035%CoCl 2 6H 2 O, 0.005~0.025%ZnSO 4 7H 2 O, 0.0015~0.0025%CuSO 4 ·5H 2 O, the balance is seawater, more preferably: 4% (NH 4 ) 2 SO 4 , 0.6~0.7%KNO 3 、1.3% CaCl 2 ·2H 2 O, 2.5% MgSO 4 7H 2 O, 3.0% K 2 HPO 4 , 2.8% KH 2 PO 4 , 0.2~0.3%FeSO 4 7H 2 O, 0.2~0.3%EDTA-Na 2, 0.01~0.15%NaMoO 4 ·2H 2 O, 0.02% MnCl 2 , 0.00015~0.00025%CoCl 2 6H 2 O, 0.015~0.020%ZnSO 4 7H 2 O, 0.0020% CuSO 4 ·5H 2 O, the remainder is sea water;

[0088] The method for expanding the culture in step ② is preferably: inoculating the bacteria into 2216E liquid culture medium at a rate of 1-5%, and culturing until 10 4 ~10 6 / ml, more preferably: the bacteria are inoculated into 2216E liquid culture medium at a 3% inoculum amount and cultured until 10 5 Pieces / ml;

[0089] The volume ratio of the nitrate-reducing Marinobacterium solution, the chemoautotrophic sulfur-oxidizing bacteria solution, the Erythrobacterium solution, the salt sediment slow-growing Bacillus solution and the seawater in step ③ is preferably 1-2:1-2:1-2:1-2:1000, and more preferably 1.5:1.5:1.5:1000;

[0090] The temperature of the continued soaking is preferably 20 to 28° C., more preferably 24° C., and the time of the continued soaking is preferably 5 to 14 days, more preferably 7 to 10 days.

[0091] In the present invention, the post-introduction management method in step (3) is preferably: feeding cage fish feed on the day of introduction, the 2nd, 4th to 5th, 7th to 8th, 10th to 12th, and 15th to 17th day; selecting 2 to 4 1m2 cages on the day of introduction, the 15th, 30th, 90th, and 150th day, respectively. 2 Excavate the seabed area, calculate the survival rate of the single-ringed spiny worm in the area on the 15th, 30th, 90th and 150th days, and carry out seedling replenishment 1 to 3 times, preferably 2 times, according to the survival situation;

[0092] The calculation formula of the retention rate is:

[0093]

[0094] When the retention rate is lower than 50%, supplementary seedlings are carried out.

[0095] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0096] Example 1

[0097] An area of ​​about 50 square meters was selected in the East Village of Daqin Island, Changdao (latitude and longitude 120.838053°E, 38.299053°N) to conduct a bacterial flora-spinys sludge treatment experiment in cage aquaculture. A control group and an experimental group were set up. The control group used 0.5g, 10mm and 1.0g, 25mm spinys sludge fed with seaweed and not fortified (named control group 1 and control group 2, respectively), and no bacterial flora was introduced. The experimental group used 1.0g, 25mm long spinys sludge seedlings for bait stress resistance and environmental stress resistance.

[0098] The method of strengthening the stress resistance of the bait is: before the monocyclic worm seedlings are introduced into the breeding area, the stress-strengthening bait is continuously fed for 10 days, and the amount of the stress-strengthening bait is 1.5% of the body weight of the monocyclic worm seedlings / day;

[0099] The bait fortification formula is as follows: 45% seaweed ground liquid (total weight), 6% fish meal, 4.57% starch, 8% ultrafinely crushed fermented soybean meal, 25% sea mud, 0.03% yeast selenium ultrafine powder, 8% scallop edge waste powder, 1% sodium alginate, 1.5% vitamin premix, 0.5% mineral premix, 0.3% taurine magnesium, 0.06% tea polyphenols, and 0.04% fermented all-trans vitamin K. After the powdered raw materials are crushed to 200 mesh, all the formulas are mixed, 30% water is added and stirred into a dough, and the dough is put into the pellet extruder, and the extrusion density is set to 1.5g / cm 3 , extruded into pellets with a diameter of 1 mm and a height of 2 mm, dried to a moisture content of less than 8%, and then crushed into fine particles of 80 mesh. Before introduction, the monocyclic stingray was continuously fed in the culture pond at an amount of 1.5% of the estimated body weight per day for 10 days.

[0100] Environmental stress resistance enhancement: mainly anti-hypoxia enhancement, that is, by adjusting the aeration volume, the dissolved oxygen is slowly reduced from 8mg / L to 2mg / L at a rate of 1mg / L per day. Environmental stress resistance enhancement is carried out simultaneously with bait stress resistance enhancement, that is, the dissolved oxygen content is slowly reduced while bait stress resistance enhancement is carried out. The duration of environmental stress resistance enhancement is 12 days, and the lowest dissolved oxygen drops to 2mg / L at the end of stress resistance enhancement.

[0101] Introduce bacterial colonies in the later stage of strengthening. Prepare sediment treatment bacterial colonies and introduce them: ① Crushing and rinsing of volcanic rock particles: volcanic rock microparticles are crushed from commercially available volcanic rock particles, with a crushing particle size of 0.4mm. After crushing, they are soaked in 2 times natural seawater for 3 days. ② Preparation of nutrient salts: Add 3% (NH4) to the seawater soaked with volcanic rock particles 2 SO 4 ,0.6%KNO 3 ,1.2%CaCl2 ·2H 2 O, 3.2% MgSO 4 7H 2 O, 2.5% K 2 HPO 4 ,1.8%KH 2 PO 4 ,0.15%FeSO 4 7H 2 O, 0.4% EDTA-Na 2 ,0.008%NaMoO 4 ·2H 2 O, 0.016% MnCl 2 ,0.00007%CoCl 2 6H 2 O, 0.015% ZnSO 4 7H 2 O, 0.0018% CuSO 4 ·5H 2 O. ③ Construction of bacterial clusters: Non-photosynthetic bacteria Marinobacter nitrate-reducing Marinobacter (commercially available, purity 100%), chemoautotrophic sulfur-oxidizing bacteria (commercially available, purity 100%), Erythrobacter flavus21-3 (commercially available, purity 100%), and salt sediment bradybacillus (commercially available, purity 100%) were inoculated into 2216E liquid culture medium at a 5% inoculation rate and gradually expanded to 10 6 4. Bacteria introduction: Collect the already constructed volcanic rock bacteria clusters and sprinkle them in the breeding area at a rate of 200g / m 2 .

[0102] The introduction of the single-ringed worm was carried out in the coastal waters with a temperature of 17°C and a current speed of about 0.02m / s (sea state 0). The introduction time was in the afternoon and the introduction density was 60 / m 2 The cages were fed with fish feed on the day of introduction, 2, 4, 7, 10, and 15 days after the introduction of the seedlings. The effect evaluation was conducted on the day (initial), 15 days, 1 month, 3 months, and 5 months after the introduction of the seedlings. Three cages with a length of 1 m 2 The substrate area of ​​a certain size was excavated and the retention rate (%) of Echinops monocyclica in the area was calculated.

[0103] Table 1 Comparison of the survival rate of Echinops monocyrthospermum in each treatment group

[0104]

[0105] Note: The data are the mean ± standard deviation of three times. Considering the low retention rate after 15 days, the seedlings were supplemented once. According to the retention rate, 60 / m 2 .

[0106] Implementation effect (Table 1): The retention rate of control group 2 was much higher than that of control group 1 from the time of seedling supplementation to 5 months. Control group 2 used large-sized seedlings while control group 1 used small-sized seedlings, indicating that seedling size has a greater impact on the retention rate and large-sized seedlings have stronger adaptability. The retention rate of the single-ringed spiny borer in the experimental group was much higher than that in control groups 1 and 2, indicating that bait enhancement and bacterial introduction can improve the retention rate of the single-ringed spiny borer at 5 months. Implementation effect ( Figures 2 to 5 ): After 5 months, the improvement of sulfide, nitrogen, carbon and phosphorus in the bottom mud of each experimental group area was measured and analyzed. It was found that the sulfide, total carbon, nitrogen and phosphorus content of control group 2 were significantly lower than those of control group 1, and the experimental group was significantly lower than the two control groups, indicating that the introduction of monocyclic schizonts and bacterial groups can improve the aquaculture bottom quality well.

[0107] Example 2

[0108] In the Laizhou Bay aquaculture area (latitude and longitude 119.8609099°E, latitude 37.2821438°N), two 900m 2 A comparative experiment was conducted in two water surface areas, namely the control area and the experimental area, which were 200 meters apart. The control area was not treated in any way, while the experimental area was stocked with Echinoceros unicyclicus and introduced with bacterial groups. The caged fish in both areas were fed normally. The experimental area used large-sized seedlings of Echinoceros unicyclicus weighing 1.3g and 28-30mm in length. Echinoceros unicyclicus was subjected to feed stress resistance and environmental stress resistance enhancement;

[0109] The method of strengthening the stress resistance of the bait is as follows: before the monocyclic worm seedlings are introduced into the breeding area, the stress-strengthening bait is continuously fed for 10 days, and the amount of the stress-strengthening bait is 1.5% / day of the body weight of the monocyclic worm seedlings. The bait strengthening formula is as follows: 30% of seaweed ground liquid (accounting for the total weight), 12% of fish meal, 10%, 5% of ultrafinely crushed fermented soybean meal, 36% of sea mud, 0.05% of yeast selenium ultrafine powder, 12% of scallop edge waste powder, 1% of sodium alginate, 1% of vitamin premix, 1% of mineral premix, 0.1% of taurine magnesium, 0.02% of tea polyphenols, and 0.08% of fermented all-trans vitamin K. After the powdered raw materials are crushed to 200 meshes, all the formulas are mixed, 30% of water is added and stirred into a dough state, and the dough is put into the particle extruder, and the extrusion density is set to reach 1.5g / cm 3 The pellets were extruded into a granular material with a diameter of 4 mm and a height of 1 mm, dried to a moisture content of less than 8%, and then crushed into fine particles of 80 mesh. Before introduction, the monocyclic spiny worms were continuously fed in the culture pond at a feeding amount of 1.5% / day of the estimated body weight for 10 days.

[0110] Environmental stress resistance enhancement: mainly anti-hypoxia enhancement, that is, by adjusting the inflation volume, the dissolved oxygen is slowly reduced from 8mg / L to 2mg / L at a rate of 1mg / L per day. Environmental stress resistance enhancement is carried out simultaneously with bait stress resistance enhancement, that is, the dissolved oxygen content is slowly reduced while bait stress resistance enhancement is carried out. The duration of environmental stress resistance enhancement is 12 days, and the lowest oxygen level drops to 2mg / L at the end of stress resistance enhancement.

[0111] Preparation of sediment treatment bacterial colonies and introduction: ① Crushing and rinsing of volcanic rock particles: The particle size of volcanic rock microparticles is 0.5 mm, and after crushing, they are soaked in 3 times natural seawater for 5 days. ② Preparation of nutrient salts: Add 4% (NH 4 ) 2 SO 4 ,0.85%KNO 3 ,1.0%CaCl 2 ·2H 2 O, 3.0% MgSO 4 7H 2 O, 2% K 2 HPO 4 ,2.2%KH 2 PO 4 ,0.1%FeSO 4 7H 2 O, 0.2% EDTA-Na 2 ,0.006%NaMoO 4 ·2H 2 O, 0.03% MnCl 2 ,0.0003%CoCl 2 6H 2 O, 0.005% ZnSO 4 7H 2 O, 0.0015% CuSO 4 ·5H 2 O. ③ Bacterial activation: Non-photosynthetic bacteria Marinobacter nitrate-reducing Marinobacter (commercially available, purity 100%), chemoautotrophic sulfur-oxidizing bacteria (commercially available, purity 100%), Erythrobacter flavus 21-3 (commercially available, purity 100%), and salt sediment bradybacillus (commercially available, purity 100%) were taken in 2216E liquid medium at a 1% inoculation amount and gradually expanded and cultured until 10 4 4. Bacteria introduction: Collect the already constructed volcanic rock bacteria clusters and sprinkle them in the breeding area at a rate of 300g / m 2 .

[0112] The introduction of Echinops monocyrta was carried out in the morning when the temperature was 20°C and the current speed was 0.01 m / s (sea state 0), and the introduction density was 45 / m 2 The cage was fed with fish feed on the day of introduction, 2, 4, 7, 10, and 15 days. The effect evaluation was conducted on the day (initial), 1, 3, and 5 months after the introduction of seedlings. Three cages with a length of 1 m 2 The substrate area of ​​the same size was excavated and the survival rate of the single-ringed worm was calculated. Two seedlings were supplemented after 1 month and 3 months respectively according to the survival rate.

[0113] Table 2 Comparison of the survival rate of Echinops monocyrta in the control area and the experimental area (%)

[0114]

[0115] Experimental results ( Figures 6 to 9 ): It can be seen that the control area was only found twice in 5 months, with one head each time. It is speculated that it is an individual existing in nature or an individual that rushed over from the experimental area, while the monocyclic schizonts in the experimental area have formed a scale and can perform bottom treatment functions. Analysis of the bottom mud shows that the sulfide, total carbon, phosphorus and nitrogen in the experimental area have all decreased to varying degrees, far lower than the control area. It is possible that the introduced bacterial mass and monocyclic schizonts can convert nitrogen, phosphorus, carbon and sulfur into other substances and remove them from the environment, indicating that the technology of the present invention can significantly improve the bottom mud quality of the aquaculture cage.

[0116] As can be seen from the above embodiments, the present invention provides a method for in-situ treatment of cage culture bottom using a bacterial flora-Schizontidae combination, comprising the following steps: (1) conducting stress-resistant and enhanced culture of Schizontidae seedlings to obtain enhanced Schizontidae seedlings; (2) introducing the enhanced Schizontidae seedlings and bacterial flora into the culture area; (3) managing the Schizontidae seedlings after introduction. The present invention adopts a pollution treatment method using a bacterial flora-Schizontidae combination based on the nutritional characteristics of the bottom sludge of offshore aquaculture, and in particular, adopts an efficient implantation method to introduce Schizontidae and bacterial flora, so that they form an ecosystem at the bottom of the aquaculture cage area. The system can be used for fish residual bait and metabolic waste, so as to achieve the purpose of purifying the aquaculture bottom, protecting the ecological environment of offshore waters, and improving economic benefits.

[0117] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for in-situ treatment of bottom sediment in cage culture by using bacterial flora and Echinops unicinctus, characterized in that: The steps include: (1) performing stress-resistant strengthening cultivation on the seedlings of the single-ringed spiny moth to obtain strengthened seedlings of the single-ringed spiny moth; (2) Introducing the enhanced Echinops monocyclica seedlings and bacterial colonies into the culture area; (3) Carry out post-introduction management of the seedlings of P. monocyrta; The stress-resistant enhancement culture in step (1) includes feed stress-resistant enhancement and environmental stress-resistant enhancement; The method of strengthening the stress resistance of the bait is: before the monocyclic worm seedlings are introduced into the breeding area, the stress-strengthening bait is continuously fed for 5 to 14 days, and the amount of the stress-strengthening bait is 1 to 2% of the body weight of the monocyclic worm seedlings per day; The stress-resistant and strengthened bait comprises the following raw materials in parts by weight: 25-45 parts of ground seaweed liquid, 3-15 parts of fish meal, 3-10 parts of starch, 5-20 parts of ultrafinely crushed fermented soybean meal, 25-45 parts of sea mud, 0.01-0.05 parts of yeast selenium ultrafine powder, 4-12 parts of scallop edge waste powder, 1-3 parts of sodium alginate, 0.5-2.0 parts of vitamin premix, 0.5-1.5 parts of mineral premix, 0.1-0.5 parts of taurine magnesium, 0.02-0.1 parts of tea polyphenols, and 0.04-0.15 parts of fermented all-trans vitamin K; The method of environmental stress strengthening is: by adjusting the aeration volume from 7-8 mg / L of dissolved oxygen to slowly decrease to 1.5-2 mg / L at a rate of 0.5-1 mg / L per day, the environmental stress strengthening is carried out simultaneously with the bait stress strengthening, and the duration of the environmental stress strengthening is 10-14 days; The method for preparing the bacterial cluster in step (2) comprises the following steps: ① After crushing the volcanic rocks, soak them in seawater for 3 to 7 days, then add nutrient salts and continue soaking for 1 to 2 days to obtain soaked volcanic rocks; ② respectively expanding and culturing the nitrate-reducing Marinobacter, the chemoautotrophic sulfur-oxidizing bacteria, the red bacillus and the salt sediment slow-growing Bacillus to obtain the nitrate-reducing Marinobacter bacterial solution, the chemoautotrophic sulfur-oxidizing bacteria bacterial solution, the red bacillus bacterial solution and the salt sediment slow-growing Bacillus bacterial solution; ③ Add the nitrate-reducing Marinobacterium bacterial solution, the chemoautotrophic sulfur-oxidizing bacteria bacterial solution, the red bacillus bacterial solution and the salt sediment slow-growing Bacillus bacterial solution to the seawater in step ①, and continue soaking to obtain a bacterial mass; In step ③, the volume ratio of the nitrate-reducing Marinobacterium solution, the chemoautotrophic sulfur-oxidizing bacteria solution, the Erythrobacterium solution, the salt sediment bradybacillus solution and the seawater is 1-2:1-2:1-2:1-2:1-2:1000.

2. The method according to claim 1, characterized in that The weight of the seedlings of the monocyclic spiny worm in step (1) is 0.3-1.0 g and the body length is 20-50 mm.

3. The method according to claim 2, characterized in that The preparation method of the stress-resistant and enhanced bait in step (1) is: A. Grinding fish meal, starch, ultrafinely pulverized fermented soybean meal, ultrafine yeast selenium powder and scallop edge waste powder into 150-200 meshes to obtain a mixed powder; B. mixing the mixed powder, ground seaweed liquid, sea mud, sodium alginate, vitamin premix, mineral premix, magnesium taurine, tea polyphenols and fermented all-trans vitamin K to obtain a mixture; C. Add water to the mixture and stir, then extrude it into pellets with a diameter of 1~4mm and a height of 1~3mm; D. drying the pellets to a moisture content of less than 8%, and then crushing them into fine particles with a particle size of 50 to 100 meshes to obtain the stress-resistant and enhanced bait; The amount of water added in step C is 10-30% of the mass of the mixture. The pellet extruder is used for extrusion, and the extrusion density during extrusion is set to 1.2-1.5 g / cm 3 .

4. The method according to claim 3, characterized in that The method for introducing the enhanced monocyclic spiny worm seedlings in step (2) is as follows: the enhanced monocyclic spiny worm seedlings are placed in the offshore waters at a distance of 0 to 20 cm from the seabed; a seedling release device is used for the introduction; the temperature during the introduction is 16 to 25° C., the sea current speed is 0 to 0.1 m / s, and the sea state is 0 to 2 levels; the density of the introduced monocyclic spiny worm seedlings is 30 to 70 per m 2 .

5. The method according to claim 4, characterized in that The method for introducing the bacterial mass in step (2) is: the bacterial mass is introduced at a concentration of 150-300 g / m 2 The standard is to spray it in the breeding area.

6. The method according to claim 1, characterized in that The particle size of the crushed volcanic rock in step ① is 0.2-0.8 mm, and the mass ratio of the volcanic rock to seawater is 1:1-3; The seawater after adding nutrient salts comprises the following raw materials by weight: 3-5% (NH4)2SO4, 0.5-0.85% KNO3, 1.0-1.6% CaCl2·2H2O, 2.0-3.0% MgSO4·7H2O, 2.0-4.0% K2HPO4, 1.8-3.8% KH2PO4, 0.1-0.4% FeSO4·7H2O, 0.2-0.4% EDTA-Na2, 0.005-0.02% NaMoO4·2H2O, 0.01-0.03% MnCl2, 0.00005-0.00035% CoCl2·6H2O, 0.005-0.025% ZnSO4·7H2O, 0.0015-0.0025% CuSO4·5H2O, and the balance is seawater; The method for expanding the culture in step ② is: inoculate the bacteria into 2216E liquid culture medium at a rate of 1-5%, and culture until 10 4 ~10 6 Pieces / ml; The temperature of the continued soaking is 20-28° C., and the time of the continued soaking is 5-14 days.

7. The method according to claim 6, characterized in that The post-introduction management method described in step (3) is: feeding cage fish feed on the day of introduction, the second day, the fourth to fifth day, the seventh to eighth day, the tenth to twelfth day, and the fifteenth to seventeenth day; selecting 2 to four 1m2 cages on the day of introduction, the fifteenth day, the 30th day, the 90th day, and the 150th day; 2 Excavate the seabed area, calculate the survival rate of Echinops monocyrthistus on the 15th, 30th, 90th and 150th days, and carry out 1 to 3 replenishment according to the survival rate; The calculation formula of the retention rate is: When the retention rate is lower than 50%, supplementary seedlings are carried out.

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