A kind of Rhodotorula marina, fermentation preparation, preparation method, fermented feed and application

Abstract

The present invention belongs to the field of microbial technology and discloses a marine red yeast, a fermentation preparation, a preparation method, a fermented feed and an application. The marine red yeast is Rhodotorula mucilaginsoa RMXS-1805, and the deposit number is: CGMCCNO.25049. The composite bacterial preparation for fermenting marine aquaculture solid waste is composed of marine red yeast, Enterococcus faecalis and Lactobacillus plantarum. The present invention screens and preserves a strain of marine red yeast, and supplements a certain carbon source and nitrogen source to the residual bait and feces produced in the marine aquaculture process through the biotechnology method of microbial fermentation, adjusts the carbon-nitrogen ratio, and then adds marine red yeast, Enterococcus faecalis and Lactobacillus plantarum, adjusts the appropriate material-water ratio and temperature, and realizes the fermentation of the residual bait and feces produced in the marine aquaculture process into a recyclable feed resource.

Description

Technical Field

[0001] The present invention belongs to the field of microbial technology, and in particular relates to marine red yeast, a fermentation preparation, a preparation method, a fermented feed and applications. Background Art

[0002] Factory-based recirculating aquaculture is rapidly developing in my country, significantly increasing its economic benefits compared to traditional outdoor earthen pond aquaculture. However, this high-density aquaculture method inevitably leads to higher levels of solid waste. This solid waste primarily consists of feces excreted by aquatic animals during the aquaculture process and uneaten leftover bait. This leftover bait is highly nutritious, with crude protein and crude fat content exceeding 26% and 14%, respectively. If this organic matter remains in the aquaculture water for extended periods, its nutrients will leach into the water, forming suspended particles. This not only causes excessive nitrogen and phosphorus levels in the aquaculture water, deteriorating water quality, but also significantly affects the gills, the respiratory organs of fish, reducing the survival and growth rates of seedlings and, in severe cases, leading to fish mortality. If leftover bait and feces in the aquaculture water are not promptly cleaned, they will deteriorate and produce toxic substances. At the same time, ammonia nitrogen, nitrite nitrogen, and other substances toxic to aquatic animals will rapidly increase in the water. When they reach a certain concentration, they will directly lead to the death of aquatic animals. Aquatic animals growing in severely polluted water will inevitably suffer serious developmental consequences, such as decreased immunity, slower growth, and increased susceptibility to various diseases. This will not only result in substandard aquatic product quality, but also a significant decrease in production, ultimately leading to severe economic losses. Given this background, the solid waste problem of factory aquaculture needs to be urgently addressed.

[0003] While the technology for fermenting livestock and poultry manure through microbial fermentation is well established, it has so far been unsuccessful in treating aquaculture solid waste, let alone marine aquaculture. Marine aquaculture solid waste contains high levels of salt, and many microorganisms that can ferment livestock and poultry manure are inhibited from growing when used in the fermentation process, resulting in poor fermentation results. Therefore, it is necessary to identify salt-tolerant bacteria for fermenting marine aquaculture solid waste.

[0004] Marine red yeast is a fungus that is widely found in the marine environment. It has strong stress resistance and can synthesize a large amount of carotenoids. It contains high protein content, vitamins, trace elements and a certain amount of unsaturated fatty acids. It also produces some growth factors and digestive enzymes during its growth process. These substances can promote the digestion and absorption of feed by aquatic animals, thereby improving the utilization rate of feed.

[0005] Existing similar technical solutions: Patent No. CN109170193A: 72-75 parts chicken manure, 15-20 parts corn flour, 12-14 parts rice bran, 6-8 parts wheat bran, 5-12 parts distiller's grains, and appropriate amount of water. The steps are as follows:

[0006] The first step is to soak the wheat bran in warm water for 30-36 hours, until the surface of the wheat bran produces a lot of bubbles and has a fragrance;

[0007] In the second step, chicken manure is poured into a cement fermentation tank, 25-40% hot water is added, and then it is sealed with a plastic film for 2-5 days. The inner layer of the film is covered with water droplets. When the chicken manure fermentation is completed, the plastic film is opened and corn flour and bran are added and stirred evenly. Then the soaked wheat bran is poured into the fermentation tank, stirred evenly, and sealed for 24-30 hours. The secondary fermentation is completed when the wheat bran aroma appears, forming a fermentation mixture.

[0008] The third step is to spray hot water while mixing the fermentation mixture and the wine lees so that the water content is neither higher than 60 parts nor lower than 55 parts. The preparation of chicken manure fermented pig feed is completed.

[0009] Through the above analysis, the problems and defects of the existing technology are: the existing technology does not add fermentation agents, but allows it to ferment naturally, the fermentation speed is relatively slow, and the fermentation process is also relatively complicated.

[0010] The difficulty in solving the above problems and defects is that it is difficult to find bacteria with good fermentation effect and suitable fermentation process conditions.

[0011] The significance of solving the above problems and defects is that the residual bait and feces can be fermented quickly and conveniently, the operability is strong, and the method can be widely promoted and applied. Summary of the Invention

[0012] To overcome the problems existing in the related art, the present invention discloses a method for preparing a marine red yeast, a fermentation preparation, a preparation method, a fermented feed and its application, and specifically relates to a method for preparing a microbial fermentation feed of aquatic animal leftover bait and feces.

[0013] The technical solution is as follows: a marine red yeast, the marine red yeast is Rhodotorula mucilaginsoa RMXS-1805, the preservation date is June 10, 2022, the preservation address is the General Microbiology Center of the China Culture Collection Administration, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, and the preservation number is CGMCC NO.25049.

[0014] Another object of the present invention is to provide an application of the marine red yeast as a salt-tolerant bacterium in the fermentation treatment of aquaculture solid waste.

[0015] Another object of the present invention is to provide a marine red yeast, wherein the method for culturing the marine red yeast fermentation preparation comprises:

[0016] The red yeast was inoculated into YPD liquid culture medium by streaking and cultured. The inoculum was then inoculated into liquid fermentation medium A at a volume ratio of 1% to obtain a fermentation preparation of the red yeast. The medium A consisted of 30-35 g glucose, 1-4 g beef extract, 1-5 g ammonium chloride, 10-15 g yeast extract, and 1000-1500 g Chenhai water, with a pH of 7-7.5.

[0017] Another object of the present invention is to provide a composite bacterial preparation for fermenting marine aquaculture solid waste using the marine red yeast. The composite bacterial preparation for fermenting marine aquaculture solid waste consists of marine red yeast, Enterococcus faecalis and Lactobacillus plantarum. The addition amount of marine red yeast is 1-6×10 9 cfu / g): Enterococcus faecalis (1~6×10 9 cfu / g: Lactobacillus plantarum (1~6×10 9 cfu / g).

[0018] Another object of the present invention is to provide an application of the composite bacterial preparation for fermenting marine aquaculture solid waste in aquaculture solid waste fermentation and marine aquaculture environment regulation.

[0019] Another object of the present invention is to provide a method for preparing the composite bacterial preparation of fermented marine aquaculture solid waste, the method for preparing the composite bacterial preparation of fermented marine aquaculture solid waste comprising:

[0020] Step 1: fermentation and cultivation of various components of the composite bacterial preparation of fermented marine aquaculture solid waste;

[0021] Step 2: Optimize the mixing ratio of each component of the composite bacteria preparation for fermenting marine aquaculture solid waste.

[0022] In one embodiment, in step 1, the fermentation culture of each component of the composite bacterial preparation for fermenting marine aquaculture solid waste includes culturing a marine red yeast fermentation preparation, culturing a plant lactobacillus and a faecal enterococcus fermentation preparation;

[0023] The cultivation of the Lactobacillus plantarum and Enterococcus faecalis fermentation preparation comprises: inoculating Lactobacillus plantarum and Enterococcus faecalis into LB liquid culture medium respectively by streaking on a plate for cultivation; then inoculating the inoculum into liquid fermentation medium B at a volume ratio of 1% to obtain the fermentation preparation of Lactobacillus plantarum and Enterococcus faecalis;

[0024] The culture medium B is composed by mass of 10g-15g of peptone, 20g-25g of glucose, 5g-10g of yeast extract, 10g-15g of beef extract, 0.1g-0.5g of magnesium sulfate, 2g-5g of dipotassium hydrogen phosphate, 0.05g-0.1g of manganese sulfate, 1ml-3ml of Tween-80, 5g-8g of sodium acetate, 2g-5g of ammonium citrate and 1000g-1500g of distilled water, with a pH of 7-7.5;

[0025] In the step 2, the bacterial solutions of Rhodotorula marineum, Lactobacillus plantarum, and Enterococcus faecalis were optimized according to the colony count as follows:

[0026] (3×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g);

[0027] (6×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g);

[0028] (3×10 9 cfu / g):(6×10 9 cfu / g):(3×10 9 cfu / g);

[0029] (3×10 9 cfu / g):(3×10 9 cfu / g):(6×10 9 cfu / g).

[0030] Another object of the present invention is to provide a fermented feed for marine aquaculture prepared using the marine red yeast. The method for preparing the fermented feed for marine aquaculture comprises the following steps:

[0031] The invention uses screened and preserved marine red yeast to supplement carbon and nitrogen sources in the residual bait and feces produced in the marine aquaculture process through microbial fermentation biotechnology, adjusts the carbon-nitrogen ratio, and then adds marine red yeast, enterococcus faecalis and lactobacillus plantarum. The feed-water ratio and temperature are adjusted to ferment the residual bait and feces produced in the marine aquaculture process into a recyclable marine aquaculture fermented feed. The carbon-nitrogen ratio is (12-14):1, the fermentation temperature is 29°C, the water content in the feed-water ratio is 55-65%, and the colony count ratio of marine red yeast, enterococcus faecalis and lactobacillus plantarum is (1-6×10 9 cfu / g): Enterococcus faecalis (1~6×10 9 cfu / g: Lactobacillus plantarum (1~6×10 9cfu / g), fermentation time 96h, preferred marine red yeast: Enterococcus faecalis: Lactobacillus plantarum = (3×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g).

[0032] Another object of the present invention is to provide an application of the marine aquaculture fermented feed in preparing shrimp aquaculture feed or Artemia aquaculture feed.

[0033] Combining all the above technical solutions, the advantages and positive effects of the present invention are as follows:

[0034] First, in view of the technical problems existing in the above-mentioned prior art and the difficulty of solving these problems, this paper closely combines the technical solutions to be protected by the present invention and the results and data during the research and development process, and analyzes in detail and in depth how the technical solutions of the present invention solve the technical problems and some creative technical effects brought about by solving the problems. The specific description is as follows:

[0035] The invention adopts a fermentation method combining marine red yeast and lactic acid bacteria to ferment the residual bait and feces in the aquaculture process, thereby increasing the nutritional value.

[0036] The colony number ratio of the composite fermentation of marine red yeast, Lactobacillus plantarum and Enterococcus faecalis is (3×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g).

[0037] The fermentation process conditions of the fermented leftover bait and feces by marine red yeast, Lactobacillus plantarum and Enterococcus faecalis of the present invention are: the carbon-nitrogen ratio (12-14): 1, the fermentation temperature is 29°C, the water content in the material-water ratio is 55-65%, preferably the carbon-nitrogen ratio is 12:1, the fermentation water content is 55%, and the fermentation time is 96h.

[0038] The nutritional value (crude protein, crude fat and amino acids) of the fermentation product of the present invention is improved, and the effect of cultivating Artemia is far better than that of a treatment group without fermentation.

[0039] Specifically, the marine red yeast of the present invention contains abundant proteins, vitamins, digestive enzymes, carbohydrates, amino acids, unsaturated fatty acids, and immune-enhancing nutritional factors. The present invention utilizes a composite fermentation agent combining marine red yeast with Lactobacillus plantarum and Enterococcus faecalis. Compared to conventional fermentation agents using only lactic acid bacteria, this composite fermentation agent has the advantage of increasing the nutritional value of the fermentation product. Furthermore, the fermentation process is simple and easy to operate.

[0040] The fermented feed was crushed and used for Artemia culture. Compared with Artemia fed directly with unfermented leftover bait and feces, the survival rate, body length and weight of Artemia fed with the microbial fermented feed were higher than those of the group fed with unfermented leftover bait and feces, and the water quality of the culture was also better than that of the group fed with leftover bait and feces.

[0041] The present invention evaluated the fermentation effect and measured the fermented products. Compared with those before fermentation, crude protein and crude fat increased, crude ash and crude fiber decreased, and total amino acid content increased.

[0042] Table 1 pH and odor changes before and after fish feces fermentation

[0043]

[0044] Table 2 Changes in conventional nutrients

[0045]

[0046] Table 3 Changes in amino acids in fermented fish feces (%)

[0047]

[0048]

[0049] Second, considering the technical solution as a whole or from the perspective of the product, the technical effects and advantages of the technical solution to be protected by the present invention are described in detail as follows:

[0050] The Lactobacillus plantarum and Enterococcus faecalis of the present invention are commercial strains purchased from Qingdao Genyuan Biotechnology Group Co., Ltd.

[0051] The present invention selects and preserves a strain of marine red yeast, and uses the biotechnology method of microbial fermentation to supplement a certain carbon source and nitrogen source to the residual bait and feces produced in the marine aquaculture process, adjust the carbon-nitrogen ratio, and then add marine red yeast, Enterococcus faecalis and Lactobacillus plantarum, adjust the appropriate material-water ratio and temperature, and realize the fermentation of the residual bait and feces produced in the marine aquaculture process into a recyclable feed resource.

[0052] When the fermentation product of the present invention is fed to aquaculture, the water quality will be better compared with directly feeding unfermented leftover bait and feces.

[0053] The purpose of the present invention is to use the biotechnology method of microbial fermentation technology to convert the leftover bait and feces produced in the aquaculture process into feed resources that can be utilized for recycling, thereby turning waste into treasure.

[0054] Third, as auxiliary evidence for the inventiveness of the claims of the present invention, it is also reflected in the following important aspects:

[0055] (1) The expected benefits and commercial value of the technical solution of the present invention after transformation are as follows: in Vannamei shrimp farming, 100 kilograms of normal commercial feed produces 75-85 kilograms of shrimp, and the cost is about RMB 800, while 100 kilograms of fecal microbial fermentation feed mixed with commercial feed in a 1:1 ratio produces 50-60 kilograms of shrimp, and the cost is about RMB 400, so it has great commercial value. In addition, the fecal microbial fermentation feed is of great significance in the commercial breeding of Artemia. Compared with the traditional feeding of Artemia with chicken manure, Artemia fed with fecal microbial fermentation feed grows faster. 100 kilograms of fecal microbial feed can produce 60-70 kilograms of Artemia, which has less pollution to water quality; while 100 kilograms of feces can produce 40-50 kilograms, but it has more serious water pollution.

[0056] (2) The technical solution of the present invention fills the technical gap in the industry at home and abroad: The technology of the present invention fills the gap in the resource utilization of solid waste in the aquaculture industry at home and abroad, which is of great significance.

[0057] (3) Whether the technical solution of the present invention solves the technical problem that people have always wanted to solve but have never been able to solve: The technical solution of the present invention solves the technical problem of resource utilization of leftover bait and feces in aquaculture.

[0058] (4) Whether the technical solution of the present invention overcomes the technical prejudice: The technical solution of the present invention overcomes the technical prejudice that microbial fermentation can only be carried out on livestock and poultry manure in the past.

[0059] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the present disclosure and, together with the description, serve to explain the principles of the present disclosure.

[0061] Figure 1 Schematic diagram of the phylogenetic tree constructed based on the 18S rDNA sequence of the strain RMXS-1805 provided in the embodiment of the present invention.

[0062] Figure 2 This is a schematic diagram of taking a bacterial liquid of marine red yeast RMXS-1805, inoculating it into YPD liquid culture medium with glucose concentrations of 10, 20, 40, 60, and 80 g / L, culturing it in a shaker at 25° C. and 160 rpm for 48 hours, and then measuring its biomass, as provided in an embodiment of the present invention.

[0063] Figure 3This is a schematic diagram of taking a bacterial liquid of Rhodotorula marineum RMXS-1805, inoculating it into a YPD liquid culture medium, culturing it at rotation speeds of 160, 180, 200, and 220 rpm for 48 hours, and then measuring its biomass, as provided in an embodiment of the present invention.

[0064] Figure 4 This is a schematic diagram of taking a bacterial liquid of Rhodotorula marineum RMXS-1805, inoculating it into YPD liquid culture medium with pH values of 4.5, 5.0, 5.5, 6.0, and 6.5, and culturing it for 48 hours, and then measuring its biomass, as provided in an embodiment of the present invention.

[0065] Figure 5 This is a schematic diagram of taking a bacterial liquid of marine red yeast RMXS-1805, inoculating it into a YPD liquid culture medium, culturing it in a shaker at 18, 22, 26, and 30° C. for 48 hours, and then measuring its biomass, as provided in an embodiment of the present invention.

[0066] Figure 6 This is a schematic diagram of taking a bacterial liquid of Rhodotorula marineum RMXS-1805, inoculating it into a YPD liquid culture medium, culturing it for 48, 72, 96, 120, and 144 hours, and then measuring its biomass, as provided in an embodiment of the present invention.

[0067] Figure 7 The present invention provides a flow chart of a method for preparing a preparation for resource utilization of marine aquaculture solid waste provided by an embodiment of the present invention.

[0068] Figure 8 This is a schematic diagram of the concentration of marine red yeast in the fermentation product in the optimized ratio of the composite bacterial preparation provided by an embodiment of the present invention.

[0069] Figure 9 This is a rendering of the product after fermentation provided by an embodiment of the present invention.

[0070] Figure 10 This is a schematic diagram of the concentration of marine red yeast in the fermentation product during the carbon-nitrogen ratio optimization provided by an embodiment of the present invention.

[0071] Figure 11 This is a schematic diagram of the concentration of marine red yeast in the fermentation product during the fermentation temperature optimization provided by an embodiment of the present invention.

[0072] Figure 12 Schematic diagram of the concentration of marine red yeast in the fermentation product during the optimization of fermentation moisture content provided by an embodiment of the present invention.

[0073] Figure 13 This is a schematic diagram of the concentration of marine red yeast in the fermentation product during the fermentation time optimization provided by an embodiment of the present invention.

[0074] Figure 14This is a schematic diagram of the changes in ammonia nitrogen during the breeding period provided by an embodiment of the present invention.

[0075] Figure 15 This is a diagram showing changes in nitrite nitrogen during aquaculture provided by an embodiment of the present invention.

[0076] Figure 16 This is a diagram of nitric nitrogen changes during aquaculture provided by an embodiment of the present invention. DETAILED DESCRIPTION

[0077] To make the above-mentioned objects, features, and advantages of the present invention more readily apparent, specific embodiments of the present invention are described in detail below with reference to the accompanying drawings. The following description sets forth numerous specific details to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways than those described herein, and those skilled in the art may make similar modifications without departing from the scope of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0078] 1. Explanatory Examples In order to enable those skilled in the art to fully understand how to implement the present invention, this section provides an illustrative example that expands upon the technical solutions of the claims.

[0079] An embodiment of the present invention provides a marine red yeast, the marine red yeast deposit number of which is: CGMCCNO.25049; the depositor is: Tian Xiangli, Ocean University of China, College of Fisheries of Ocean University of China, No. 5 Yushan Road, Shinan District, Qingdao City, Shandong Province; the deposit date is: June 10, 2022; the deposit address is: General Microbiology Center of China Culture Collection Administration, Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; the biological material is: RMXS-1805; the classification name is: Glue red yeast Rhodotorula mucilaginsoa, and the test result on June 10, 2022 was survival.

[0080] The present invention also provides a composite bacterial preparation for fermenting marine aquaculture solid waste using the marine red yeast. The composite bacterial preparation for fermenting marine aquaculture solid waste is composed of marine red yeast, Enterococcus faecalis and Lactobacillus plantarum, and the colony count ratio is marine red yeast: Enterococcus faecalis: Lactobacillus plantarum = marine red yeast (1-6×10 9 cfu / g): Enterococcus faecalis (1~6×10 9 cfu / g: Lactobacillus plantarum (1~6×10 9 cfu / g). Among them, Enterococcus faecalis and Lactobacillus plantarum are commercial strains obtained from Qingdao Genyuan Biotechnology Group Co., Ltd.

[0081] The embodiment of the present invention further provides a method for preparing a composite bacterial preparation of fermented marine aquaculture solid waste, comprising: fermenting and culturing the components of the composite bacterial preparation of fermented marine aquaculture solid waste;

[0082] Optimization of the mixing ratio of each component of the composite bacterial preparation for fermentation of marine aquaculture solid waste.

[0083] The embodiment of the present invention further provides a fermented feed for seawater aquaculture prepared using the marine red yeast. The method for preparing the fermented feed for seawater aquaculture comprises the following steps:

[0084] The invention uses screened and preserved marine red yeast to supplement carbon and nitrogen sources in the residual bait and feces produced in the marine aquaculture process through microbial fermentation biotechnology, adjusts the carbon-nitrogen ratio, then adds marine red yeast, enterococcus faecalis and lactobacillus plantarum, adjusts the material-water ratio and temperature, and ferments the residual bait and feces produced in the marine aquaculture process into a recyclable marine aquaculture fermented feed; the carbon-nitrogen ratio is (12-14):1, the fermentation temperature is 29°C, the water content in the material-water ratio is 55-65%, and the total colony count ratio of marine red yeast, enterococcus faecalis and lactobacillus plantarum is marine red yeast: enterococcus faecalis: lactobacillus plantarum = marine red yeast (1-6×10 9 cfu / g): Enterococcus faecalis (1~6×10 9 cfu / g: Lactobacillus plantarum (1~6×10 9 cfu / g), fermentation time 96h.

[0085] In the embodiment of the present invention, the carbon-nitrogen ratio is 12:1, the fermentation temperature is 29°C, and the water content in the material-water ratio is 55%;

[0086] Or use a carbon-nitrogen ratio of 14:1, a fermentation temperature of 29°C, and a water content of 65% in the material-water ratio;

[0087] Or use a carbon-nitrogen ratio of 13:1, a fermentation temperature of 29°C, and a water content of 60% in the material-water ratio.

[0088] The technical solution of the present invention is further described below in conjunction with different embodiments.

[0089] Example 1 Screening and Isolation of a Marine Rhodotorula Strain

[0090] Isolation and purification of marine red yeast: A portion of the sediment sample collected from the redfin pufferfish breeding pond was added to a sterile YPD liquid culture medium and cultured at 25°C for 3 days. Then, 1 ml of the above bacterial culture solution was accurately pipetted into a sterile centrifuge tube and graded diluted with sterile physiological saline to prepare 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10-6 , 10 -7 , 10 -8 Use a pipette to draw 0.2 ml of the dilution solution and evenly spread it on YPD solid culture medium. Then place it in a 28°C incubator in the dark and culture for 72 hours. After a single colony grows on the plate, use an inoculation loop to pick up a suspected (orange or red) single colony on the plate and streak it on the YPD solid culture medium to further obtain a high-purity single colony. After isolation and purification, the obtained single colony is stored at 4°C for future use.

[0091] Example 2 Classification and Identification of a Marine Rhodotorula

[0092] 1. Morphological identification of marine red yeast: The bacteria are single-celled, oval or spherical, without flagella, with a cell diameter of approximately 3 to 8 μm, no pseudohyphae, and reproduce by budding; the colonies of this strain are red or orange-red, with a diameter of 3 to 5 mm, a smooth surface, a raised center, opaque, easy to pick up, and neat edges.

[0093] 2. Physiological and biochemical characteristics of marine red yeast: The physiological and biochemical characteristics of marine red yeast are shown in Table 4:

[0094] Table 4 Physiological and biochemical characteristics of marine red yeast strain RMXS-1805

[0095]

[0096] 3. Molecular genetic characteristics

[0097] DNA from Rhodotorula marineensis RMXS-1805 was extracted and amplified using universal primers (Shanghai Sangon Biotechnology Co., Ltd.) ITS1: 5'-TCCGTAGGTGAACCTGCGG-3' and ITS4: 5'T-CCTCCGCTTATTGATATGC-3'. The PCR reaction system consisted of 2 μL template DNA, 1 μL ITS1 primer, 1 μL ITS4 primer, 25 μL Taq enzyme, and 1 μL ddH2O. The PCR reaction procedure included 30 cycles of initial denaturation (95°C for 5 min), denaturation (95°C for 45 s), annealing (56°C for 45 s), and extension (72°C for 30 s), followed by a final incubation at 72°C for 10 min. The PCR amplification products were detected by agarose gel electrophoresis. After passing the test, they were sequenced at Qingdao BGI Biotechnology Co., Ltd. and the final sequencing results were entered into the website: www.NCBI.nlm.gow. Click on the BLAST analysis software to compare the measured gene sequence with the sequence in the GenBank database. Finally, the Neighbor-Joining method in MEGA7 was used to construct a phylogenetic tree. The results showed that it was highly similar to the sequence of Rhodotorula glutinis (such as Figure 1The strain RMXS-1805 was finally determined to belong to the genus Rhodotorula mucilaginosa (shown in the phylogenetic tree constructed from the 18S rDNA sequence of the strain RMXS-1805).

[0098] Example 3 Determination of Growth Conditions of Marine Red Yeast RMXS-1805

[0099] 1. Take the Rhodotorula RMXS-1805 bacterial suspension and inoculate it into YPD liquid medium with glucose concentrations of 10, 20, 40, 60 and 80 g / L. Incubate it in a shaker at 25°C and 160 rpm for 48 hours, and then measure its biomass. Figure 2 As shown in the figure, the biomass was the highest when the glucose concentration was 40 g / L, so 40 g / L was selected as the optimal concentration.

[0100] 2. Take the Rhodotorula RMXS-1805 bacterial solution, inoculate it into YPD liquid medium, culture it at the speed of 160, 180, 200, and 220 rpm for 48 hours, and then measure its biomass. Figure 3 As shown, when the rotation speed is 200 rpm, the biomass is the highest, so 200 rpm is selected as the optimal rotation speed.

[0101] 3. Take the Rhodotorula marineis RMXS-1805 bacterial suspension and inoculate it into YPD liquid medium with pH values of 4.5, 5.0, 5.5, 6.0, and 6.5, respectively, and culture for 48 hours. Then measure its biomass. Figure 4 As shown in the figure, the biomass was the highest when the pH was 4.5, so pH 4.5 was selected as the optimal culture pH.

[0102] 4. Take the marine red yeast RMXS-1805 bacterial liquid, inoculate it into YPD liquid medium, place it in a shaker at 18, 22, 26, and 30℃ for 48 hours, and then measure its biomass. Figure 5 As shown in the figure, when the temperature is 26℃, the biomass is the highest, so 26℃ is selected as the optimal culture temperature.

[0103] 5. Take the marine red yeast RMXS-1805 bacterial liquid, inoculate it into YPD liquid medium, culture it for 48, 72, 96, 120, and 144 hours, and then measure its biomass. Figure 6 As shown in the figure, the biomass was the largest when the culture time was 48 h, so 48 h was selected as the optimal culture time.

[0104] In summary, the experiments showed that R. maritima R. Rhodotorula RMXS-1805 grew best at a glucose concentration of 40 g / L, a rotation speed of 200 rpm, a pH of 4.5, a temperature of 26°C, and a growth time of 48 h.

[0105] Example 4

[0106] like Figure 7 As shown, the preparation method of the marine aquaculture solid waste resource utilization preparation provided by the embodiment of the present invention includes:

[0107] S101, Optimization of formula of composite bacterial preparation for fermentation of aquaculture solid waste.

[0108] Strains: Rhodotorula marineum (RMXS-1805), Lactobacillus plantarum (ZW), Enterococcus faecalis (FC).

[0109] (1) Cultivation of R. mutans (RMXS-1805) fermentation preparation: R. mutans was inoculated into YPD liquid medium by streaking on a plate and cultured at 26°C and 200 r / min for 48 h. Then, 1% by volume of the inoculum was inoculated into liquid fermentation medium A and cultured at 26°C and 200 r / min for 48 h to obtain R. mutans (RMXS-1805) fermentation preparation. At this time, the concentration of the fermentation preparation could reach 10 8 cfu / ml.

[0110] The culture medium A is composed of 30-35 g of glucose, 1 g-4 g of beef extract, 1 g-5 g of ammonium chloride, 10 g-15 g of yeast extract and 1000-1500 g of Chenhai water by mass, and has a pH of 7-7.5.

[0111] (2) Fermentation preparation culture of Lactobacillus plantarum (ZW) and Enterococcus faecalis (FC): Lactobacillus plantarum (ZW) and Enterococcus faecalis (FC) were inoculated into LB liquid culture medium by streaking on plates and cultured at 28°C in the dark for 24-48 hours; then, they were inoculated into liquid fermentation medium B at a volume ratio of 1% and cultured at 28°C and 180 r / min for 48 hours to obtain fermentation preparations of ZW and FC. At this time, the concentration of the fermentation preparations could reach 10 9 cfu / ml.

[0112] The culture medium B is composed by mass of 10g-15g of peptone, 20g-25g of glucose, 5g-10g of yeast extract, 10g-15g of beef extract, 0.1g-0.5g of magnesium sulfate, 2g-5g of dipotassium hydrogen phosphate, 0.05g-0.1g of manganese sulfate, 1ml-3ml of Tween-80, 5g-8g of sodium acetate, 2g-5g of ammonium citrate and 1000g-1500g of distilled water, with a pH of 7-7.5;

[0113] S102, optimization of the formula ratio of compound bacterial preparations.

[0114] The cultured bacterial solutions of marine red yeast RMXS-1805, Lactobacillus plantarum ZW, and Enterococcus faecalis FC were mixed in the following proportions (number of colonies):

[0115] (3×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g);

[0116] (6×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g);

[0117] (3×10 9 cfu / g):(6×10 9 cfu / g):(3×10 9 cfu / g);

[0118] (3×10 9 cfu / g):(3×10 9 cfu / g):(6×10 9 cfu / g).

[0119] Then, the mixture is mixed to make four composite bacterial preparations with different proportions, which are then used as fermentation agents and mixed with leftover bait and feces for subsequent fermentation.

[0120] A fixed mass of 25g of fish feces (containing 10g of dry matter) was placed in each sample bag, and the corresponding bran was added to adjust the carbon-nitrogen ratio to 12:1. The moisture was controlled at 55%, and 4 composite bacterial preparations with different ratios were added. Three parallel preparations were set up for each ratio. After adding the bacterial solution, the whole system was mixed and then placed in a sealed bag and sealed. It was then placed in a constant temperature incubator at 26°C for fermentation for 96 hours. After fermentation, the optimal formula of the composite bacterial preparation was determined by recording the sensory effects of the fermentation products and combining them with the concentration of marine red yeast.

[0121] Experimental results:

[0122] (1) Sensory effects of fermentation products, see Table 5.

[0123] Table 5 Sensory effects of fermentation products

[0124] Compound bacteria ratio Color and sensory smell <![CDATA[(3×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g)]]> Bright color, with a strong wine lees and grassy flavor <![CDATA[(6×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g)]]> Brighter in color, with a stronger wine lees and grassy aroma <![CDATA[(3×10 9 cfu / g):(6×10 9 cfu / g):(3×10 9 cfu / g)]]> Dark in color, with a strong wine lees and grassy smell <![CDATA[(3×10 9 cfu / g):(3×10 9 cfu / g):(6×10 9 cfu / g)]]> Dark in color, with a strong wine lees and grassy smell

[0125] (2) In the optimization of the ratio of compound bacterial preparation, the concentration of marine red yeast in the fermentation product is shown in Figure 8 .

[0126] The comprehensive sensory fermentation effect and the concentration of marine red yeast can be seen that the composite bacteria ratio is (3×10 9 cfu / g):(3×10 9cfu / g):(3×10 9 cfu / g) can be used as the optimal ratio. Figure 9 This is a rendering of the product after fermentation provided by an embodiment of the present invention.

[0127] Example 5

[0128] In a preferred embodiment of the present invention, the fermented feed preparation method provided by the present invention includes: by screening and preserving a strain of marine red yeast, supplementing a certain carbon source and nitrogen source to the leftover bait and feces produced in the marine aquaculture process through the biotechnology method of microbial fermentation, adjusting the carbon-nitrogen ratio, and then adding marine red yeast, Enterococcus faecalis and Lactobacillus plantarum, adjusting the appropriate feed-water ratio and temperature, thereby achieving the fermentation of the leftover bait and feces produced in the marine aquaculture process into a recyclable feed resource.

[0129] Specifically include:

[0130] Optimization process of fermentation feed process conditions.

[0131] Collect the leftover bait and feces from the factory-scale recirculating aquaculture pond, drain the excess water (leave it for about 24 hours), and finally concentrate the water content of the leftover bait and feces to about 60% (probably the state where the leftover bait and feces can naturally form). The carbon-nitrogen ratio of the leftover bait and feces is about 10:1.

[0132] A. Carbon-nitrogen ratio optimization:

[0133] Each sample bag was filled with a fixed mass of 25g of fish feces (containing 10g of dry matter), and then the corresponding amount of bran was added according to the set carbon-nitrogen ratio. An appropriate amount of water was added to control the moisture of the system at 55%, and the carbon-nitrogen ratio was set to five gradients of 10:1, 11:1, 12:1, 13:1, and 14:1, with 3 replicates for each gradient. Then, marine red yeast RMXS-1805, Lactobacillus plantarum ZW, and Enterococcus faecalis FC were cultured at a rate of (3×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g) ratio, and after adding the bacterial solution, the whole system was mixed, then placed in a sealed bag, sealed, and placed in a constant temperature incubator at 26°C for fermentation for 96 hours. After fermentation, the optimal carbon-nitrogen ratio of the fermentation substrate was comprehensively determined by recording the sensory effects of the fermentation products and combining them with the concentration of marine red yeast.

[0134] Experimental results:

[0135] The sensory effects of fermentation products are shown in Table 6.

[0136] Table 6 Sensory effects of fermentation products

[0137] Carbon-nitrogen ratio Color and sensory smell 10：1 Black in color and has a strong odor 11：1 The color is dark brown, with a bad smell and a little wine lees smell 12：1 Bright yellow in color, odorless, with a strong wine lees and grassy aroma 13：1 Bright yellow in color, odorless, with a strong wine lees and grassy aroma 14：1 Bright yellow in color, odorless, with a strong wine lees and grassy aroma

[0138] (2) The concentration of marine red yeast in the fermentation product during the optimization of carbon-nitrogen ratio is shown in Figure 10 .

[0139] A comprehensive analysis of the sensory fermentation effects and Rhodotorula concentrations revealed that bacterial concentrations increased with increasing carbon-nitrogen ratios. Furthermore, the odor of the fermented product decreased until it became odorless and gradually shifted to a wine lees and grassy aroma. The C-N ratio of 10:1, without bran, produced a foul-smelling, darkly colored fermentation product. However, the C-N ratio of 12:1, 13:1, and 14:1 was odorless, with a wine lees and grassy aroma and a brighter color, but no significant differences were observed. Considering fermentation costs, the C-N ratio of 12:1, with a relatively low amount of bran added, was the optimal ratio.

[0140] B. Fermentation temperature optimization.

[0141] Each sample bag was filled with a fixed mass of 25 g of fish feces (containing 10 g of dry matter), and the corresponding bran was added to adjust the carbon-nitrogen ratio to 12:1. The moisture content was controlled at 55%, and the fermentation temperature was set at 20°C, 23°C, 26°C, 29°C, and 32°C. There were three replicates for each gradient. Then, RMXS-1805, Lactobacillus plantarum ZW, and Enterococcus faecalis FC were cultured at a temperature of (3 × 10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g) ratio, and after adding the bacterial solution, the whole system was mixed, then placed in a sealed bag, sealed, and placed in a corresponding constant temperature incubator for fermentation for 96 hours. After fermentation, the optimal fermentation temperature was comprehensively determined by recording the sensory effects of the fermentation products and combining the concentration of marine red yeast.

[0142] Experimental results:

[0143] (1) Sensory effects of fermentation products, see Table 7.

[0144] Table 7 Sensory effects of fermentation products

[0145] Temperature Color and sensory smell 20 Black in color and has a strong odor 23 Dark in color, with a strong odor 26 The color is brown, the odor is not obvious, and it has a little wine lees smell and grass fragrance. 29 Yellow in color, odorless, with a strong wine lees smell and grassy aroma 32 The color is yellow-brown, odorless, and the wine lees smell and grassy aroma are lighter than those at 29℃

[0146] (2) The concentration of marine red yeast in the fermentation product during fermentation temperature optimization is shown in Figure 11 .

[0147] A comprehensive analysis of the sensory fermentation effects and the concentration of Rhodotorula spp. showed that at 29°C, the fermentation product had a strong lees and grassy flavor. At the same time, the concentration of Rhodotorula spp. reached its highest level at 29°C. Therefore, 29°C was selected as the optimal fermentation temperature.

[0148] C. Optimization of fermentation moisture content.

[0149] Each sample bag was filled with a fixed mass of 25 g of fish feces (containing 10 g of dry matter), and the corresponding bran was added to adjust the carbon-nitrogen ratio to 12:1. The moisture content was set to 35%, 45%, 55%, and 65% in four gradients, with three replicates for each gradient. Then, R. oxysporum RMXS-1805, L. plantarum ZW, and Enterococcus faecalis FC were added according to the colony count (3 × 10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g) ratio, and after adding the bacterial solution, the whole system was mixed, then placed in a sealed bag, sealed, and placed in a constant temperature incubator at 29°C for 96 hours. After fermentation, the optimal fermentation moisture content of the fermentation substrate was comprehensively determined by recording the sensory effects of the fermentation products and combining them with the concentration of marine red yeast.

[0150] Experimental results:

[0151] (1) Sensory effects of fermentation products, see Table 8.

[0152] Table 8 Sensory effects of fermentation products

[0153] Moisture content% Color and sensory smell 35 Black in color, with a strong odor, and clumping and shriveling 45 The color is dark, there is a strong odor, and there are lumps and shriveling phenomena. 55 Bright color, no odor, relatively fluffy and moist, with a strong wine lees smell and grassy aroma 65 Darker in color, smelly, and thinner fermentation products

[0154] (2) The concentration of marine red yeast in the fermentation product during the optimization of fermentation moisture content, e.g. Figure 12 shown.

[0155] Comprehensively considering the sensory fermentation effect and the concentration of marine red yeast, it can be seen that when the moisture content is 55%, the sensory effect of the fermentation product is the best and the concentration of marine red yeast reaches the highest. Therefore, the moisture content of 55% is selected as the optimal fermentation moisture content.

[0156] D. Fermentation time optimization.

[0157] A fixed mass of 25 g of fish feces (containing 10 g of dry matter) was placed in each sample bag, and the corresponding bran was added to adjust the carbon-nitrogen ratio to 12:1. The moisture was controlled at 55%, the fermentation temperature was 29 °C, and the fermentation time was set to 48 h, 72 h, 96 h, and 120 h, for a total of 5 time periods, with 3 replicates in each time period. Then, marine red yeast RMXS-1805, Lactobacillus plantarum ZW, and Enterococcus faecalis FC were added according to the colony count (3 × 10 9 cfu / g):(3×10 9 cfu / g):(3×10 9cfu / g) was added, and after adding the bacterial solution, the whole system was mixed, then placed in a sealed bag, sealed, and placed in a constant temperature incubator at 29°C for fermentation. After fermentation, the optimal fermentation time was determined comprehensively by recording the sensory effects of the fermentation products and combining the concentration of marine red yeast.

[0158] Experimental results

[0159] (1) Sensory effects of fermentation products, see Table 9.

[0160] Table 9 Sensory effects of fermentation products

[0161] Fermentation time h Color and sensory smell 48 Dark in color, with a strong odor 72 The color is brown, the odor is not obvious, and it has a little wine lees smell and grass fragrance. 96 Brown in color, basically odorless, with a strong wine lees smell and grassy aroma 120 Brown in color, basically odorless, with a strong wine lees smell and grassy aroma

[0162] (2) The concentration of marine red yeast in the fermentation product during fermentation time optimization is shown in Figure 13 .

[0163] Based on the comprehensive sensory fermentation effect and the concentration of marine red yeast, it can be seen that 96h can be used as the optimal fermentation time.

[0164] In summary, the fermentation scheme is: carbon-nitrogen ratio 12:1, fermentation temperature 29°C, water content 55%, composite bacteria ratio (3×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g), fermentation time 96h.

[0165] In a preferred embodiment of the present invention, after step S103, a fermentation effect evaluation is required: a fixed mass of 25 g of fish feces (containing 10 g of dry matter) is placed in each sample bag, the corresponding bran is added to adjust the carbon-nitrogen ratio to 12:1, the moisture is controlled at 55%, the fermentation temperature is 29°C, the fermentation time is 96 h, three parallels are set, and then the marine red yeast RMXS-1805, Lactobacillus plantarum ZW and Enterococcus faecalis FC are added according to the colony count (3×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g) was added, the bacterial solution was added, and the entire system was mixed thoroughly. The mixture was then sealed in a sealed bag and placed in a 29°C constant temperature incubator for fermentation. The fermented product was dried in a dryer and crushed to a 60-mesh size using a grinder to obtain a microbial fermented feed for Artemia cultivation. Measurements of the fermented product showed increases in crude protein and crude fat compared to pre-fermentation levels, decreases in crude ash and crude fiber, and an increase in total amino acid content compared to pre-fermentation levels.

[0166] Table 10 pH and odor changes before and after fish feces fermentation

[0167]

[0168] Table 11 Changes in conventional nutrients

[0169] Before fermentation After fermentation crude protein 23.32％ 25.38％ crude fat 9.85％ 10.15％ Crude ash 14.62％ 13.46％ crude fiber 7.52％ 6.44％

[0170] Table 12 Changes in amino acids in fermented fish feces (%)

[0171]

[0172]

[0173] Example 7

[0174] The present invention provides a composite bacterial preparation for fermenting marine aquaculture solid waste using the marine red yeast. The composite bacterial preparation for fermenting marine aquaculture solid waste consists of marine red yeast, Enterococcus faecalis and Lactobacillus plantarum. The colony count of marine red yeast: Enterococcus faecalis: Lactobacillus plantarum is (3×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g).

[0175] Example 8

[0176] The present invention also provides a method for preparing a composite bacterial preparation of fermented marine aquaculture solid waste, comprising:

[0177] Step 1: screening and isolation of marine red yeast strains, classification and identification of a marine red yeast strain, molecular genetic characterization, and determination of growth conditions of marine red yeast;

[0178] Step 2, fermentation culture of each component of the composite bacteria preparation of fermented marine aquaculture solid waste;

[0179] Step 3: Optimize the mixing ratio of each component of the composite bacteria preparation for fermenting marine aquaculture solid waste.

[0180] 2. Application Examples: In order to demonstrate the creativity and technical value of the technical solution of the present invention, this section provides application examples of the claimed technical solution on specific products or related technologies.

[0181] The residual bait and feces in the factory circulating aquaculture pond were collected, and the excess water was drained (left for about 24 hours), and finally the water content of the residual bait and feces was concentrated to about 60% (probably the state where the residual bait and feces can be naturally formed). The carbon-nitrogen ratio of the residual bait and feces was about 9:1. Bran was added to adjust the carbon-nitrogen ratio to 12:1, and an appropriate amount of water was added to control the moisture content of the system to about 55%. Then, marine red yeast RMXS-1805, Lactobacillus plantarum ZW and Enterococcus faecalis FC were added according to (3×10 9 cfu / g):(3×109 cfu / g):(3×10 9 After adding the bacterial solution, the whole system was mixed thoroughly, then sealed in a sealed bag and placed in a constant temperature incubator at 29°C for 96 hours. After fermentation, the fermented product was dried and ground with a 60-mesh grinder to obtain the microbial fermented feed.

[0182] Application Example 1

[0183] Farming of Litopenaeus vannamei: The normal shrimp feed of the control group was named DO group, and the dried and crushed microbial fermentation feed was mixed with the normal shrimp feed in a ratio of 1:1 and named D1 group.

[0184] The specific steps are as follows: Healthy Litopenaeus vannamei shrimp were purchased from Qingdao Baorong Aquatic Products Co., Ltd., and the salinity of the culture water was approximately 17‰. They were acclimated and temporarily cultured in aquaria. During the temporary culture period, the salinity of the culture water was increased by 2‰ daily to approximately 30‰, and stabilized for two days before the culture experiment. After the temporary culture period, the shrimp were starved for 24 hours. Then, 150 healthy, uniformly sized shrimp (initial weight 3.57±0.01g) were selected and evenly distributed into 10 50L glass aquariums, with 15 shrimp placed in each aquarium. The 10 glass aquariums were randomly divided into two experimental groups, with five replicates in each group. During the 42-day culture experiment, the shrimp were fed twice daily, at 8:00 am and 5:00 pm, with a daily feed amount of 4%-5% of their body weight. Before each feeding, leftover bait and feces were siphoned out. Change the water once a day, with the water replacement volume being about 1 / 3 to 2 / 3 of the original water level. The water temperature is around 23°C, the salinity is 28‰-30‰, the pH is 7.8-8.0, and the dissolved oxygen content is above 5 mg / L. Check and record the shrimp feeding and mortality every day.

[0185] After the experiment, the shrimp were starved for 24 hours, and their final weight was weighed and recorded. The following growth indicators were also calculated:

[0186] Survival rate = (number of shrimp after the experiment / number of shrimps stocked before the experiment) × 100%

[0187] Weight gain rate (WGR) = (W 末 -W 初 )×100% / W 初

[0188] Specific growth rate (SGR) = [(lnW 末 -lnW 初 ) / t]×100%

[0189] Where: W 末 —End weight, W 初 —Initial weight

[0190] The experimental results are shown in Table 13:

[0191] Table 13 Growth, specific growth rate, survival rate and weight gain rate of Penaeus vannamei

[0192]

[0193] Note: Different letters indicate significant differences among them (P<0.05).

[0194] From the growth data in the table, it can be seen that compared with the normal feed group, adding part of the fecal microbial fermentation feed to the feed does not affect the growth of Vannamei shrimp. Therefore, in practical aquaculture production, part of the fecal microbial fermentation feed can be added to the normal feed to save aquaculture costs and at the same time achieve the purpose of recycling waste resources.

[0195] Application Example 2

[0196] Artemia culture: The dried and crushed leftover bait and feces are named feed D0 group, and the dried and crushed microbial fermentation feed group is named D1.

[0197] The specific steps are as follows: 6 glass tanks (35×35cm) were divided into 2 groups, each with 3 tanks, namely group D0 and group D1. 100 Artemia larvae of the same size were placed in each glass tank. The same amount of unfermented leftover bait and feces and microbial fermented feed were fed to group D0 and group D1 at 8:00, 13:00, and 19:00 every day for 15 days. The bottom of the tank was cleaned once a day. The water was not changed during the whole process. The ammonia nitrogen NH4 in the water was measured every 3 days. + -N, nitrite NO2 - -N, nitrate NO3 - -N content was recorded. After the rearing was completed, all Artemia in each tank were fished out and counted separately to calculate their survival rates. Ten Artemia were randomly selected from each tank and weighed and measured to calculate the average weight and average length. Finally, the water quality indicators after the rearing were measured. The results are shown in Table 14 below:

[0198] Table 14 Growth of Artemia

[0199] Treatment group Survival rate / % Body length / mm Body weight / mg D0 <![CDATA[47.88% a ]]> <![CDATA[2.95 a ]]> <![CDATA[12.19 a ]]> D1 <![CDATA[83.65% b ]]> <![CDATA[4.23 b ]]> <![CDATA[17.56 b ]]>

[0200] Changes in ammonia nitrogen during the breeding period are shown in Figure 14 For the changes of nitrite nitrogen during the breeding period, see Figure 15 For the changes of nitrate and nitrogen during the breeding period, see Figure 16 .

[0201] from Figure 14-16From the data, we can see that the survival rate, body length and weight of Artemia in the D1 group fed with microbial fermented feed were better than those in the D0 group fed with unfermented leftover bait and feces; and the water quality was also better than that in the D0 group fed directly with unfermented leftover bait and feces.

[0202] In summary, the fermented feed provided by the present invention is crushed and used for Artemia culture. Compared with the Artemia cultured by directly feeding unfermented leftover bait and feces, the survival rate, body length and weight of Artemia cultured by feeding the microbial fermented feed are higher than those of the group fed with unfermented leftover bait and feces, and the culture water quality is also better than that of the group fed with leftover bait and feces.

[0203] The above description is only a preferred specific implementation method of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications, equivalent substitutions and improvements made by any technician familiar with this technical field within the technical scope disclosed by the present invention and within the spirit and principles of the present invention should be covered by the scope of protection of the present invention.

Claims

1. A marine red yeast, characterized in that The marine red yeast is red yeast ( Rhodotorula mucilaginsoa )RMXS-1805, the deposit date is June 10, 2022, and the deposit number is CGMCC NO. 25049.

2. Use of the marine red yeast as claimed in claim 1 as a salt-tolerant bacterium in the fermentation treatment of aquaculture solid waste.

3. A marine red yeast fermentation preparation obtained by fermentation treatment of aquaculture solid waste according to claim 2, characterized in that: The culturing method of the marine red yeast fermentation preparation comprises: The marine red yeast was inoculated into a YPD liquid medium by streaking and cultured; then, the inoculum was inoculated into a liquid fermentation medium A at a volume ratio of 1% to obtain a fermentation preparation of the marine red yeast; The culture medium A consists of 30g-35g of glucose, 1g-4g of beef extract, 1g-5g of ammonium chloride, 10g-15g of yeast extract and 1000g-1500g of Chenhai water by mass, and has a pH value of 7-7.

5.

4. A composite bacterial preparation of fermented marine aquaculture solid waste using the marine red yeast of claim 1, characterized in that: The composite bacterial preparation for fermenting marine aquaculture solid waste is composed of marine red yeast, Enterococcus faecalis and Lactobacillus plantarum, and the addition ratio according to the number of colonies is: marine red yeast: Enterococcus faecalis: Lactobacillus plantarum = (3×10 9 cfu / g):(3×10 9 cfu / g): (3×10 9 cfu / g).

5. Use of the composite bacterial preparation for fermenting marine aquaculture solid waste as claimed in claim 4 in fermentation of aquaculture solid waste and regulation of marine aquaculture environment.

6. A method for preparing a composite bacterial preparation of fermented marine aquaculture solid waste as claimed in claim 4, characterized in that: The preparation method of the composite bacterial preparation of fermented marine aquaculture solid waste comprises: Step 1: fermentation and cultivation of various components of the composite bacterial preparation of fermented marine aquaculture solid waste; Step 2: Optimize the mixing ratio of each component of the composite bacteria preparation for fermenting marine aquaculture solid waste.

7. The method for preparing the composite bacterial preparation of fermented marine aquaculture solid waste according to claim 6, characterized in that: In step 1, the fermentation culture of each component of the composite bacterial preparation for fermenting marine aquaculture solid waste includes the culture of marine red yeast fermentation preparation, the culture of plant lactobacillus and the culture of enterococcus faecalis fermentation preparation; The culture of the Lactobacillus plantarum and Enterococcus faecalis fermentation preparation comprises: inoculating Lactobacillus plantarum and Enterococcus faecalis into LB liquid culture medium by streaking a plate for culturing; then inoculating the inoculum into liquid fermentation medium B at a volume ratio of 1% to obtain the fermentation preparation of Lactobacillus plantarum and Enterococcus faecalis; the culture medium B comprises, by mass, 10g-15g of peptone, 20g-25g of glucose, 5g-10g of yeast extract, 10g-15g of beef extract, 0.1g-0.5g of magnesium sulfate, 2g-5g of dipotassium hydrogen phosphate, 0.05g-0.1g of manganese sulfate, 1ml-3ml of Tween-80, 5g-8g of sodium acetate, 2g-5g of ammonium citrate, and 1000g-1500g of distilled water, with a pH of 7-7.5; In the step 2, the bacterial solutions of Rhodotorula marineum, Lactobacillus plantarum, and Enterococcus faecalis were optimized according to the colony count as follows: (3×10 9 cfu / g):(3×10 9 cfu / g):(3×10 9 cfu / g)。 8. A fermented feed for marine aquaculture prepared using the marine red yeast of claim 1, characterized in that: The method for preparing fermented feed for marine aquaculture comprises the following steps: The invention uses screened and preserved marine red yeast to supplement carbon and nitrogen sources in the residual bait and feces produced in the marine aquaculture process through microbial fermentation biotechnology, adjusts the carbon-nitrogen ratio, and then adds marine red yeast, Enterococcus faecalis and Lactobacillus plantarum. The feed-water ratio and temperature are adjusted to ferment the residual bait and feces produced in the marine aquaculture process into a recyclable marine aquaculture fermented feed. The carbon-nitrogen ratio is (12-14):1, the fermentation temperature is 29°C, the water content in the feed-water ratio is 55%-65%, the marine red yeast, Enterococcus faecalis and Lactobacillus plantarum are added according to the colony number, and the marine red yeast (3×10 9 cfu / g): Enterococcus faecalis (3×10 9 cfu / g): Lactobacillus plantarum (3×10 9 cfu / g), fermentation time 96h.

9. Use of the fermented marine aquaculture feed according to claim 8 in preparing shrimp aquaculture feed or Artemia aquaculture feed.

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