A fish farming additive

Abstract

The present invention relates to a fish farming additive, comprising a boron-resistant Lysinibacillus boronitolerans bacterial body and / or a boron-resistant Lysinibacillus fermentation liquid; wherein the boron-resistant Lysinibacillus was deposited in the China Center for Type Culture Collection, referred to as CCTCC, on October 8, 2019, with a deposit number of CCTCC NO.M 2019773. The present application uses probiotics as a fish farming additive, which is simple to use, low in cost, green, pollution-free, residue-free, and non-toxic and side-effect-free. It can effectively improve the intestinal flora of fish, improve the immunity and disease resistance of fish, and the growth performance, and contribute to the body growth of fish.

Description

Technical Field

[0001] The invention relates to aquaculture, in particular to a fish farming additive. Background Art

[0002] In recent years, China's aquaculture industry has flourished, with aquaculture output accounting for nearly 70% of the world's aquaculture output. The total output has increased from 2.33 million tons in 1978 to 51.42 million tons in 2016, making China the only country in the world where aquaculture output exceeds wild catch.

[0003] The current intensive and dense aquaculture methods provide the best transmission pathway for pathogens. The pathogens that cause diseases in aquatic animals are mainly bacteria, viruses, fungi, parasites, and some algae. Common fish diseases caused by bacteria include gill rot, white skin, red skin, vertical scales, bacterial sepsis, bacterial enteritis, etc. When fish are infected with pathogens, they may suffer from weight loss or even mass death. Some pathogens that can cause diseases among humans, fish, and livestock are even more threatening to human health.

[0004] Therefore, improving fish immunity, disease resistance and growth performance is the primary issue that needs to be addressed at present. Summary of the invention

[0005] In view of the technical problems existing in the prior art, the present invention proposes a fish farming additive, comprising boron-resistant Lysinibacillus boronitolerans bacteria and / or boron-resistant Lysinibacillus boronitolerans fermentation broth; wherein the boron-resistant Lysinibacillus boronitolerans was deposited in the China Center for Type Culture Collection, abbreviated as CCTCC, on October 8, 2019, with a preservation number of CCTCC NO.M 2019773.

[0006] The fish farming additive as described above is a feed additive or a water additive.

[0007] The fish farming additive as described above, wherein the feed additive is formulated to be mixed into feed.

[0008] The fish farming additive as described above, wherein the boron-resistant lysine Bacillus bacteria are effective live bacteria, and the number of effective live bacteria mixed into every 1g of feed is 107 CFU / g.

[0009] The fish feed additive as described above, wherein the weight ratio (w / v) of the feed to the boron-resistant lysine bacillus fermentation liquid is (1-30):3, preferably 10:3

[0010] The fish feed additive as described above, wherein the boron-resistant lysine bacillus fermentation liquid is a boron-resistant lysine bacillus cultured to OD 600 The fermentation broth is 4.0-5.5; further, the boron-resistant lysine Bacillus fermentation broth is the boron-resistant lysine Bacillus cultured to OD 600 It is the fermentation broth at 4.7-5.0.

[0011] Application of boron-resistant lysine bacillus cells and / or boron-resistant lysine bacillus fermentation liquid in the preparation of fish farming additives.

[0012] A fish breeding method comprises adding the above-mentioned fish breeding additive into the breeding water or feed of the fish.

[0013] In the above-mentioned breeding method, the fish breeding additive is a feed additive, and the daily feeding amount of the feed additive is 5-15% of the fish body weight, preferably 7-10% of the fish body weight.

[0014] In the breeding method as described above, the fish breeding additive is a feed additive, and the feeding frequency of the feed additive is twice a day.

[0015] The present application uses probiotics as fish farming additives, which are simple to use, low-cost, green, pollution-free, residue-free, and non-toxic and side-effect-free. They can effectively improve the intestinal flora of fish, enhance the immunity, disease resistance, and growth performance of fish, and contribute to the growth of fish. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 A relative abundance diagram of tilapia intestinal microbial phylum level according to one embodiment of the present application;

[0017] Figure 2 A relative abundance diagram of tilapia intestinal microorganisms at the genus level according to one embodiment of the present application; and

[0018] Figure 3 This is a relative abundance diagram of tilapia intestinal microorganism species level according to one embodiment of the present application. DETAILED DESCRIPTION

[0019] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0020] In the following detailed description, reference may be made to the various specification drawings that are part of the present application and are used to illustrate specific embodiments of the present application. In the accompanying drawings, similar reference numerals describe substantially similar components in different figures. The various specific embodiments of the present application are described in sufficient detail below so that a person of ordinary skill in the art with relevant knowledge and skills in the art can implement the technical solutions of the present application. It should be understood that other embodiments may also be used or structural and logical changes may be made to the embodiments of the present application.

[0021] Today's intensive and dense aquaculture methods provide the best transmission pathway for pathogens. In order to combat the rampant spread of pathogens, people have adopted many methods, the most common of which is the use of antibiotics to inhibit common pathogens. However, since antibiotics can inhibit the growth of many bacteria, many beneficial bacteria in water and fish will also be indiscriminately inhibited and killed; and antibiotics will remain in water and animal bodies and be absorbed by animals, making farmed aquatic products fail to meet national drug testing standards. Existing studies have shown that antibiotics remaining in fish can pose risks to human health (Li Le et al. Study on the risk ranking of antibiotic residues in farmed tilapia [J]. China Fisheries Quality and Standards, Vol. 5, No. 5, September 2015). Many countries have banned the use of antibiotics in aquaculture.

[0022] In order to reduce the abuse of antibiotics in aquaculture, people are looking for effective alternative solutions. Joint experts from the Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization define probiotics as "live microorganisms that, when accumulated to a certain amount, benefit the health of the host" based on health assessments.

[0023] This application uses tilapia as a model animal to verify the effects of boron-resistant lysine Bacillus and / or its products on fish immunity, disease resistance and growth performance. However, tilapia is not a limited fish species for the fish feed additive of this application, and feeding other non-tilapia fish with boron-resistant lysine Bacillus and / or its products is also within the scope of protection of this application. According to one embodiment of the present invention, tilapia fry were purchased from the Wenbifeng breeding base of Hainan Baolu Aquatic Technology Co., Ltd., with an initial average body weight of 1.11±0.26g.

[0024] According to one embodiment of the present application, tilapia are divided into 4 groups, namely: a control group fed with a basic feed; a YS11 group fed with fish feed mixed with boron-resistant lysine Bacillus bacteria in the basic feed; a FB group fed with fish feed mixed with boron-resistant lysine Bacillus fermentation liquid in the basic feed; and a FLF group fed with fish feed mixed with boron-resistant lysine Bacillus bacteria and fermentation liquid in the basic feed. According to one embodiment of the present application, each group has 50 tilapia fry. According to one embodiment of the present application, the breeding environment of tilapia is the laboratory of the Agricultural Science Building of Hainan University, and the tilapia is bred in a glass fish tank. According to one embodiment of the present application, the water used to breed tilapia is tap water. In some embodiments, before each use of water, it is placed in a fish tank overnight to remove residual chlorine in the tap water.

[0025] The terms used herein have the following meanings:

[0026] The term "fish farming additive" or "farming additive" refers to a substance added to the water body or the feed of fish during the fish farming process. Antibiotics added to the water body in the prior art are a type of farming additive. In the present application, boron-resistant lysine Bacillus cells and / or their fermentation liquid are used as fish farming additives. In some embodiments, the fish farming additive is directly added to the water body. In other embodiments, the fish farming additive is mixed into the fish feed. In some embodiments, tilapia is used as a representative fish to verify the effect of fish farming additives on fish.

[0027] The term "basic feed" refers to a basic feed with fish meal and soybean meal as protein sources, supplemented with rapeseed meal, wheat, flour, monocalcium phosphate, vitamin A, vitamin D3, vitamin E, niacinamide, D-calcium pantothenate, ferrous sulfate, manganese sulfate, zinc sulfate, and copper sulfate, with a crude protein content of 42% and a crude fat content of 6%.

[0028] The term "feed additive" refers to a small or trace amount of substances added during the production, processing and use of feed. The amount used in the feed is very small but the effect is significant. For example, antibiotics are used as feed additives to inhibit the spread of pathogens in fish farming environments.

[0029] In some embodiments, boron-resistant lysine bacillus is used as a feed additive to improve the immunity, disease resistance and growth performance of fish and reduce the damage of pathogens to fish. In some embodiments, the feed additive is a live bacterial cell of boron-resistant lysine bacillus. In some embodiments, the feed additive is a fermentation liquid of boron-resistant lysine bacillus. In some embodiments, the feed additive is a mixture of live bacterial cells of boron-resistant lysine bacillus and a fermentation liquid of boron-resistant lysine bacillus.

[0030] The term "probiotics" refers to a class of active microorganisms that are beneficial to the host. Probiotics colonize in the host's intestines and reproductive system, and can produce definite health effects, thereby improving the host's microecological balance and exerting beneficial effects on the intestine. The beneficial bacteria or fungi in the human body and animals mainly include: Clostridium butyricum, lactic acid bacteria, bifidobacterium, Lactobacillus acidophilus, actinomycetes, yeast, etc.

[0031] Because probiotics adhere to intestinal epithelial cells and form a biological barrier with the intestine, the mechanism of action of probiotics in the body is as described below:

[0032] 1. Inhibit intestinal pathogens

[0033] The bacterial community in the intestine combines with the intestinal mucosa to form a biological barrier, and prevents the invasion of harmful bacteria, viruses and antigens through space occupancy effect, nutritional competition and various metabolites and bacteriocins secreted by them, stimulating the immune organs of the intestine to exert their immune functions. The intestinal tract adheres to the intestinal mucosa, and after colonization, it continues to multiply, forming a stable microbial community, reducing the invasion of conditional pathogens and preventing the spread of conditional pathogens.

[0034] 2. Regulate intestinal microbial flora

[0035] Intestinal bacteria play an important role in maintaining the host's normal physiology, nutrition, health, homeostasis, and defense against pathogens and immune system function. Once the intestinal tissue is damaged, the probability of invasion by opportunistic pathogens and viruses will increase. Probiotics can change the intestinal microbial structure of animals, increase the abundance of beneficial bacteria, and make Firmicutes and Proteobacteria the dominant bacteria in the intestine. Reduce the abundance of pathogens such as Staphylococcus, Streptococcus, Aeromonas, etc.

[0036] 3. Enhance Immunity

[0037] Probiotics improve the immunity of aquatic organisms and enhance the expression of nonspecific factors by enhancing the activity of macrophages or natural killer cells, regulating the secretion of immunoglobulins or cytokines, or indirectly by strengthening the intestinal epithelial barrier, changing mucus secretion and competitively excluding other pathogenic bacteria.

[0038] The administration method of probiotics is simple, and it can be directly delivered into the water body or mixed into the feed to feed the animals. Boron-resistant lysine bacillus is a probiotic that has an antagonistic effect on some pathogens. The present invention adds it to the basic feed of tilapia, and after long-term feeding of probiotic feed, the growth performance, intestinal digestion ability and liver immunity are enhanced.

[0039] In the present application, probiotic boron-resistant lysine bacillus and / or its product is added to fish basic feed as a feed additive. In the present application, the type of boron-resistant lysine bacillus is not limited. In certain embodiments, the boron-resistant lysine bacillus (Lysinibacillus boronitolerans) of the present application is deposited in China Center for Type Culture Collection on October 8, 2019, referred to as CCTCC, and the deposit number is CCTCC NO.M 2019773.

[0040] The term "OD value" is an abbreviation for optical density, which indicates the light density absorbed by the test substance. 600 The optical density at 600 nm can be used to indicate the bacterial cell density, where the absorbance is proportional to the concentration of bacteria in the culture medium.

[0041] The term "LB medium" is abbreviated as Luria-Bertani. LB medium is a commonly used bacterial culture medium and a semi-synthetic culture medium.

[0042] The term "weight sampling" means that the water outside the tilapia is absorbed with absorbent paper and then the weight is measured on a weighing machine.

[0043] The term "visceral sampling" means opening the abdominal cavity of the tilapia near the cloaca, taking out the entire liver and the entire intestine, flushing the blood and connective tissue on the tissue surface and the intestinal contents with PBS pre-cooled at 4°C, absorbing the water with absorbent paper and weighing it with a weighing instrument.

[0044] The term "liver disruption" means that the external connective tissue and blood are rinsed with PBS, mixed with 0.9% saline, 5-8 1.5 mm steel beads are added, and the liver is homogenized and disrupted in an oscillating disruptor at 4°C. After the supernatant is aspirated, the activity of each enzyme is determined according to the kit.

[0045] In some embodiments, kits for assaying the activities of visceral catalase (CAT), superoxide dismutase (SOD), acid phosphatase (ACP), and alkaline phosphatase (AKP) were purchased from Suzhou Keming Biotechnology Co., Ltd., China, and the operating steps followed the instructions provided with the kits.

[0046] The term "average weight gain" in this application refers to recording the average weight of tilapia on day 0 of the experiment, recorded as the initial average weight; after feeding for 70 days in the manner of Examples 1-3, weight sampling was performed to obtain the average weight of tilapia in each group after 70 days, recorded as the final average weight. Average weight gain = final average weight - initial average weight.

[0047] The term "weight gain rate %" or "WGR" in this application refers to the average weight gain percentage of tilapia in each group after 70 days of feeding according to the example relative to the weight gain before feeding, and the calculation formula is: weight gain rate % (WGR) = (final average weight - initial average weight) / initial weight × 100%.

[0048] The term "specific growth rate %" or "SGR" in this application is calculated as Specific Growth Rate % (SGR) = [(ln final average weight - ln initial average weight) / number of days of feeding] x 100%.

[0049] The term "feed to meat ratio" or "FCR" in this application refers to the relationship between the total feed consumed and the total weight gain of tilapia, and the calculation formula is: feed to meat ratio (FCR) = total weight of feed consumed / total weight gain.

[0050] The term "survival rate %" or "SR" in this application refers to the relationship between the number of tilapia initially selected and the number of tilapia surviving after 70 days of feeding in each group of experiments, and the calculation formula is: Survival rate % (SR) = final tail number / initial tail number × 100%.

[0051] The term "relative abundance" refers to the number of species in a community. The greater the species abundance, the more complex its structure and the greater its resistance stability. The fish farming additive of the present application can change the abundance of fish intestinal microbial flora, thereby regulating the intestinal microbial flora.

[0052] The present application takes a strain of boron-resistant lysine Bacillus, which is a potential antagonistic biocontrol bacterium preserved in the laboratory and can antagonize a variety of pathogenic bacteria and pathogenic fungi, as a research object. According to an embodiment of the present application, the boron-resistant lysine Bacillus can promote the growth of tilapia and promote the development of the liver and intestines.

[0053] The following examples use feed additives as fish farming additives to illustrate the technical effects of the present application. The feed additives are prepared with reference to the process flow of the following examples:

[0054] Example 1 Group YS11: Boron-resistant lysine Bacillus cells were mixed into the basic fish feed as a feed additive

[0055] 1. Pick a single colony of boron-resistant Bacillus lysine in 5 mL of LB liquid medium and culture it in a 37°C constant temperature incubator at 150 rpm overnight. 600 =0.02 was transferred to 50 mL of fresh LB liquid medium and cultured for 24 h to obtain a fresh boron-resistant lysine Bacillus culture liquid.

[0056] 2. Use a spectrophotometer to determine the concentration of the bacterial solution, dilute it with fresh LB liquid, adjust the concentration of the bacterial solution to 107 CFU, then put the liquid into a centrifuge tube, centrifuge it at 6000 rpm for 15 minutes, take the precipitate, and discard the supernatant.

[0057] 3. Mix the centrifuged cells of the above-mentioned boron-resistant lysine bacillus with LB liquid culture medium, and then mix the mixed bacterial solution with the basic feed in a sterile container. The effective viable count of boron-resistant lysine bacillus per 1g of feed is determined to be 107 CFU / g. The feed weight to liquid ratio (w / v) is 10:3. Ensure that the bacterial solution is completely mixed with the feed. After mixing, pack the feed in sterile plastic bags and store it in a refrigerator at 4°C.

[0058] 4. Feed the tilapia with the mixed feed for 70 days, with a daily feeding amount of 7-10% of the fish body weight, and feed twice a day, at 9 am and 4 pm respectively, so that the tilapia can completely eat the feed.

[0059] Change the water once a day, changing 50% of the water, and clean up the feces in the fish tank.

[0060] 5. Take samples on the 0th day after the start of culture and the 70th day after the end of culture.

[0061] 6. Take body weight and viscera samples. Disintegrate the viscera and measure superoxide dismutase (SOD), alkaline phosphatase (AKP), acid phosphatase (ACP), and catalase (CAT).

[0062] Example 2 FB group: mixing the fermentation liquid of boron-resistant Bacillus lysine into the basic feed of fish

[0063] 1. Preparation of boron-resistant lysine Bacillus fermentation broth. Inoculate the boron-resistant lysine Bacillus in LB liquid culture medium and culture it in a constant temperature incubator at 37°C and 150rpm for 96h. Then divide it into 50mL centrifuge tubes and centrifuge it at 4°C and 12000rpm for 30 minutes. Centrifuge the bacteria completely to remove the precipitate and filter the supernatant with a 0.22μm filter membrane to obtain a sterile fermentation filtrate. Store the fermentation broth at 4°C. In some embodiments, the boron-resistant lysine Bacillus fermentation broth is cultured to OD 600 The fermentation liquid after centrifugation and removal of bacteria when the OD is 4.0-5.5. In some embodiments, the fermentation liquid of the boron-tolerant lysine bacillus is cultured to an OD of 600 The fermentation liquid after centrifugation and removal of bacteria at 4.7-5.0.

[0064] 2. Mix the above-mentioned boron-resistant lysine Bacillus fermentation liquid with the basic feed, and the feed weight to liquid ratio (w / v) is 10:3. Ensure that the bacterial liquid is completely mixed with the feed. After mixing, pack the feed in sterile plastic bags and store it in a refrigerator at 4°C.

[0065] 3. Feed the tilapia with the mixed feed for 70 days, with a daily feed amount of 7-10% of the fish weight, twice a day, at 9 am and 4 pm, so that the tilapia can eat the feed completely. Change the water once a day, with a water change of 50%, and clean the feces in the fish tank.

[0066] 4. Take samples on the 0th day after the start of culture and the 70th day after the end of culture.

[0067] 5. Take body weight and viscera samples. Disintegrate the viscera and measure superoxide dismutase (SOD), alkaline phosphatase (AKP), acid phosphatase (ACP), and catalase (CAT).

[0068] Example 3 FLF group: Boron-resistant lysine Bacillus cells and fermentation liquid were mixed into the basic feed of fish

[0069] 1. Pick a single colony of boron-resistant Bacillus lysine and place it in 5 mL of LB liquid medium and culture it overnight in a 37°C constant temperature incubator at 150 rpm. 600 =0.02 was transferred to multiple 50 mL of fresh LB liquid culture medium and cultured for 24 h to obtain fresh boron-resistant lysine Bacillus bacterial liquid.

[0070] The concentration of the bacterial solution was measured using a spectrophotometer. In some embodiments, the OD value of the boron-tolerant lysine Bacillus solution in each culture medium was 600 is 4.0-5.5; in some embodiments, the OD of the boron-resistant lysine Bacillus solution in each culture medium is 600 It is 4.7-5.0.

[0071] 2. In some embodiments, the above-mentioned boron-resistant lysine Bacillus liquid is placed in a centrifuge tube, centrifuged at 6000rpm for 15min, and the precipitate A is taken, and the supernatant B is removed. In some embodiments, the liquid is placed in a centrifuge tube, centrifuged at 6000rpm for 15min, and the supernatant B' is taken, and the precipitate A' is removed. Among them, the precipitate is the bacterial body, and the supernatant is the fermentation liquid. Then the remaining precipitate A is mixed with the supernatant B' to form a new bacterial liquid. Dilute with fresh LB liquid, adjust the bacterial liquid concentration to 107CFU, and set aside.

[0072] In some embodiments, the boron-resistant lysine Bacillus bacterial solution is not centrifuged, but diluted with fresh LB liquid to adjust the bacterial solution concentration to 107 CFU for standby use.

[0073] 3. Mix any bacterial solution obtained in step 2 with the basic feed in a sterile container. The mixing ratio is 107 CFU / g of boron-resistant lysine Bacillus per 1g of feed. The feed weight to liquid ratio (w / v) is 10:3. Ensure that the bacterial solution is completely mixed with the feed. After mixing, pack the feed in sterile plastic bags and store in a refrigerator at 4°C.

[0074] 4. Feed the tilapia with the mixed feed for 70 days, with a daily feed amount of 7-10% of the fish weight, twice a day, at 9 am and 4 pm, so that the tilapia can eat the feed completely. Change the water once a day, with a water change of 50%, and clean the feces in the fish tank.

[0075] 5. Take samples on the 0th day after the start of culture and the 70th day after the end of culture.

[0076] 6. Take body weight and viscera samples. Disintegrate the viscera and measure superoxide dismutase (SOD), alkaline phosphatase (AKP), acid phosphatase (ACP), and catalase (CAT).

[0077] Example 4 LB group: LB liquid medium was mixed into the fish basal feed

[0078] 1. Mix the sterilized LB liquid medium with the basic feed, with the feed weight to liquid ratio (w / v) being 10:3. Ensure that the liquid is completely mixed with the feed, and after mixing, pack the feed in sterile plastic bags and store in a 4°C refrigerator.

[0079] 2. Feed the tilapia with the mixed feed for 70 days, with a daily feed amount of 7-10% of the fish weight, twice a day, at 9 am and 4 pm, so that the tilapia can eat the feed completely. Change the water once a day, with a water change of 50%, and clean the feces in the fish tank.

[0080] 3. Take samples on the 0th day after the start of culture and the 70th day after the end of culture.

[0081] 4. Take body weight and viscera samples. Disintegrate the viscera and measure superoxide dismutase (SOD), alkaline phosphatase (AKP), acid phosphatase (ACP), and catalase (CAT).

[0082] Example 5 Growth performance and liver immune enzyme activity of tilapia

[0083] Below, the preferred embodiment of the present invention will be further described in detail in conjunction with the following table and accompanying drawings, wherein the tilapia in the control group were fed only with the basic feed:

[0084] Table 1 shows the growth performance changes of tilapia fed with boron-resistant lysine Bacillus cells and fermentation liquid mixed with basic feed according to an embodiment of the present application, wherein the final body weight of the FB group and the FLF group was significantly higher than that of the control group and the LB group (p<0.05), the weight gain rate of the FB group and the FLF group was significantly higher than that of the control group and the LB group (p<0.05), and the feed-to-meat ratio of the FB group and the FLF group was significantly lower than that of the control group and the LB group (p<0.05). The specific growth rate of the FB group and the FLF group was significantly higher than that of the control group and the LB group (p<0.05). After adding additives to the feed, the survival rate of tilapia was improved. There was no significant difference in the liver weight of tilapia between the groups, and in the intestinal weight results, the FLF group was significantly higher than the control group and the LB group (p<0.05).

[0085] Table 1 Growth performance of tilapia

[0086]

[0087] Table 2 is the result data of liver immune enzyme activity of tilapia fed with boron-resistant lysine Bacillus cells mixed with fermentation liquid and basic feed according to one embodiment of the present application. The enzyme activities of catalase (CAT), superoxide dismutase (SOD), acid phosphatase (ACP), and alkaline phosphatase (AKP) were measured, and the results showed that the various indicators of the YS11 group, FB group, and FLF group were higher than those of the control group and LB group.

[0088] Table 2 Results of tilapia liver immune enzyme activity

[0089]

[0090] Example 6 Fish farming additives regulate the intestinal microbial flora of tilapia

[0091] In this example, the relative abundance of intestinal microorganisms of tilapia in the above examples 1-3 was measured before and after 70 days of feeding the aquaculture additives. The results are shown in Figure 1-Figure 3 :

[0092] At door level ( Figure 1), Proteobacteria, Actinobacteria, Firmicutes, and Fusobacteria constituted the four most important bacterial phyla, followed by Bacteroidetes, Acidobacteria, Verrucomicrobia, Patescibacteria, Tenericutes, and Chloroflexi. Among them, Actinobacteria was the dominant bacterial community in the control group, while Proteobacteria, Firmicutes, and Fusobacteria were the dominant bacterial communities in the experimental group.

[0093] At the genus level ( Figure 2 ), uncultured bacteriummicrobacteriaceae, bosea, uncultured bacteriumrhizobiaceae, and cetobacterium are the main bacterial genera, and also include uncultured bacterium rhizobiales, shinella, plesiomonas, uncultured bacterium muribaculaceae, leucobacter, and paracoccus.

[0094] At the species level ( Figure 3), uncultured bacteriummicrobacteriaceae, uncultured bacterium bosea, uncultured bacterium rhizobiaceae, uncultured bacteriumcetobacterium as the main bacterial species. Also included are uncultured bacteriumrhizobiales, uncultured bacterium shinella, uncultured bacteriumplesiomonas, uncultured bacteriummuribaculaceae, uncultured bacteriumleucobacter, and uncultured bacteriumparacoccus.

[0095] The above embodiments are only used to illustrate the present invention, but not to limit the present invention. Ordinary technicians in the relevant technical field can make various changes and modifications without departing from the scope of the present invention. Therefore, all equivalent technical solutions should also fall within the scope of the present invention.

Claims

1. A tilapia farming additive comprising boron-resistant lysine bacillus ( Lysinibacillus Boronitolerans ) bacteria and / or boron-resistant lysine bacillus fermentation broth; wherein, The boron-resistant lysine Bacillus was deposited in the China Center for Type Culture Collection, abbreviated as CCTCC, on October 8, 2019, with a deposit number of CCTCC NO.M2019773.

2. The tilapia breeding additive according to claim 1, wherein the additive is a feed additive or a water additive.

3. The tilapia farming additive according to claim 2, wherein the feed additive is configured to be mixed into feed.

4. The tilapia farming additive according to claim 3, wherein: Boron-resistant lysine Bacillus is an effective living bacteria, and the number of effective living bacteria mixed in 1g of feed is 10 7 CFU / g.

5. The tilapia breeding additive according to claim 3, wherein: The weight ratio (w / v) of feed and boron-resistant lysine bacillus fermentation broth is (1-30):

3.

6. The tilapia breeding additive according to claim 5, wherein: The weight ratio (w / v) of feed to boron-resistant lysine Bacillus fermentation broth is 10:

3.

7. The tilapia breeding additive according to claim 5 or 6, wherein the boron-resistant lysine bacillus fermentation liquid is a boron-resistant lysine bacillus cultured to OD 600 The fermentation liquid is 4.0-5.

5.

8. The tilapia breeding additive according to claim 7, wherein the boron-resistant lysine bacillus fermentation liquid is a boron-resistant lysine bacillus cultured to OD 600 It is the fermentation broth at 4.7-5.

0.

9. Use of boron-resistant lysine Bacillus cells and / or boron-resistant lysine Bacillus fermentation liquid in the preparation of tilapia breeding additives; wherein: The boron-resistant lysine Bacillus was deposited in the China Center for Type Culture Collection, abbreviated as CCTCC, on October 8, 2019, with a preservation number of CCTCC NO.M 2019773; the application includes improving the growth performance of tilapia.

10. A method for breeding tilapia, comprising adding the tilapia breeding additive according to claim 1 to breeding water or feed of tilapia.

11. The culture method according to claim 10, wherein: The tilapia breeding additive is a feed additive, and the daily feeding amount of the feed additive is 5-15% of the fish body weight.

12. The breeding method according to claim 11, wherein the daily feeding amount of the feed additive is 7-10% of the fish body weight.

13. The culture method according to claim 10, wherein: The tilapia breeding additive is a feed additive, and the feeding frequency of the feed additive is twice a day.

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