A method for raising ducks without resistance

Abstract

The application relates to an antibiotic-free breeding method of waterfowls, in particular to an antibiotic-free breeding method of ducks. The method comprises the following steps: (1) preparation of fermented feed of ducks: duck-source compound probiotics are used to ferment full-price feed of ducks, and the prepared fermented feed is used as daily feed at 100%; (2) addition of beneficial bacteria in drinking water: A. addition of duck-source compound probiotics in drinking water; B. addition of compound beneficial microbial metabolites in drinking water; (3) sterilization of litter: breeding excrement and high-temperature fermentation bacteria are added to carry out high-temperature harmless fermentation treatment on the litter in the duck breeding process, and the high temperature above 70 DEG C lasts for 5 days. The application can improve the absorption of feed, improve the immunity of ducks, guarantee the environmental protection and recycling in breeding, and solve the environmental protection problem in breeding.

Description

TECHNICAL FIELD

[0001] The application relates to an antibiotic-free breeding method of waterfowls, in particular to an antibiotic-free breeding method of ducks. BACKGROUND

[0002] Excessive use of antibiotics, improper medication scheme, and transduction of bacterial drug resistance genes, etc. result in poorer and poorer clinical use effect of antibiotics. Long-term and large-scale use of antibiotics in waterfowl breeding causes accumulation of antibiotics in the waterfowl, which brings great harm to food safety and human health. Therefore, the antibiotic-free breeding method is urgently needed in the current waterfowl breeding process, and has dual significance of reality and future development. SUMMARY

[0003] The application aims to provide an antibiotic-free breeding method of ducks.

[0004] The application is implemented as follows:

[0005] The antibiotic-free breeding method of ducks comprises the following steps:

[0006] (1) Preparation of fermented feed of ducks

[0007] Duck compound probiotics are used to ferment duck complete feed, and the prepared fermented feed is 100% used as daily feed for feeding;

[0008] (2) Drinking water with added beneficial bacteria

[0009] A. Drinking water with added duck compound probiotics;

[0010] B. Drinking water with added compound beneficial microbial metabolites;

[0011] Specific drinking is as follows:

[0012] 1-5 days old, duck compound probiotic drinking water prevention; 11-14 days old, compound beneficial microbial metabolite drinking water prevention; 16-20 days old, duck compound probiotic drinking water prevention; 23-26 days old, compound beneficial microbial metabolite drinking water prevention; the above cycle drinking, 8 hours of drinking water is provided per day;

[0013] (3) Bedding sterilization

[0014] Feces and high-temperature fermentation bacteria are added to carry out high-temperature harmless fermentation treatment on the bedding in the duck breeding process, and the high temperature above 70 DEG C lasts for 5 days.

[0015] Further, the preparation method of the duck-derived composite probiotics is as follows: taking the gastrointestinal contents of healthy ducks, increasing bacteria in BHI liquid medium, then streaking on BHI agar plate, proliferating overnight at 30-35 DEG C, picking single colonies to BHI liquid medium for proliferation, finally extracting strain DNA, PCR amplification, sequencing identification, mixing the identified beneficial bacteria for fermentation to prepare duck-derived composite probiotics.

[0016] Further, the mixed fermentation is as follows: proliferating the beneficial bacteria (making the viable bacterial count of each type of bacteria reach 1.0 x 10 8 CFU / mL), mixing, then inoculating into culture medium at an inoculation ratio of 2%, and fermenting at 33 DEG C for 72 h, and the culture medium includes the following raw materials in a weight ratio: peptone 3 g / L, yeast extract 2 g / L, glucose 3 g / L, sodium chloride 2 g / L, to prepare duck-derived composite probiotics.

[0017] Further, the composite beneficial microorganisms in the composite beneficial microbial metabolites include Levilactobacillus brevis ZG-7, Lactobacillus plantarum, Bacillus, yeast, and Bifidobacterium, wherein the Levilactobacillus brevis ZG-7 is Levilactobacillus brevis ZG-7, which has been preserved in the Guangdong Microbial Culture Collection Center (GDMCC) on June 14, 2023, with a preservation number of GDMCC No. 63562, and the rest are common beneficial bacteria.

[0018] Further, the preparation method of the composite beneficial microbial metabolites is as follows: proliferating Levilactobacillus brevis ZG-7, Lactobacillus plantarum, Bacillus, yeast, and Bifidobacterium at a weight ratio of 3:2:2:2:1 (each type of bacteria needs to be expanded and cultured to a viable bacterial count of 1.0 x 10 8 CFU / mL before mixing), proliferating overnight at 30-35 DEG C, and the culture medium includes the following raw materials in a weight ratio: peptone 8 g / L, yeast extract 5 g / L, glucose 5 g / L, sodium chloride 2 g / L, to obtain the metabolites.

[0019] Further, the bedding in step (3) is sterilized by mixing the duck breeding bedding, feces and auxiliary materials at a mass ratio of 6:3:1, and adding 0.01% of high-temperature resistant fermentation bacteria for fermentation.

[0020] The present application has the following advantages:

[0021] (1) Different duck-derived composite probiotics can be prepared for different duck varieties, and duck-derived composite probiotics are used to ferment feed for feeding different varieties of ducks, which can improve the absorption of feed.

[0022] (2) The metabolic products produced by the culture of the complex beneficial microorganisms can inhibit common harmful pathogenic bacteria such as Escherichia coli, Salmonella, Riemerella anatipestifer and Pasteurella anatipestifer in duck breeding, and improve the immunity of the duck.

[0023] (3) The high-temperature harmless fermentation treatment of the litter reduces the residues of harmful bacteria and viruses in the litter, reduces the discharge of breeding manure and wastewater, ensures the environmental protection and recycling of breeding, and solves the environmental protection problem of breeding. DETAILED DESCRIPTION

[0024] I. Materials

[0025] The general beneficial bacteria: Lactobacillus plantarum BNCC187897, Bacillus sp. BNCC221996, yeast BNCC357965, and Bifidobacterium BNCC185341 were purchased from Beina Biological Technology Co., Ltd.

[0026] The high-temperature fermentation bacteria: Bacillus sp. BNCC139195 was purchased from Beina Biological Technology Co., Ltd.

[0027] Levilactobacillus brevis ZG-7 was disclosed in patent CN202311228607.0.

[0028] II. Antibiotic-free breeding method of duck

[0029] 1. Preparation of fermented feed for duck

[0030] The beneficial complex microbial agent (containing nearly 100 beneficial bacteria including Bacillus sp., Enterococcus faecalis, Bacteroides, yeast, Bifidobacterium and Lactobacillus) isolated from the intestinal tract of duck was mixed and fermented, and the complex fermentation agent was prepared. The fermented feed for duck was prepared by adding 0.1% of the complex fermentation agent to the full-price feed for duck, and then performing solid-state feed fermentation production under the temperature and humidity conditions referring to the comprehensive pH and temperature of the bacterial community (generally 33℃, humidity 18%, fermentation time 3d).

[0031] Table 1 Effect of fermentation of different bacteria on pre-digestion of feed

[0032]

[0033] 2. Metabolic products of microorganisms

[0034] Levilactobacillus brevis ZG-7, Lactobacillus plantarum, Bacillus sp., yeast, and Bifidobacterium were mixed and cultured at a weight ratio of 3:2:2:2:1, and then proliferated overnight at 30-35℃ (the culture medium was composed of proteose peptone, yeast extract, glucose, and sodium chloride at a weight ratio of 8:5:5:2). The metabolic products obtained mainly included bacteriocin, lactic acid, acetic acid, phenyllactic acid, and formic acid, etc.

[0035] 3. Disease prevention and control during breeding

[0036] 1-5 days old, duck-derived complex probiotics 0.1% drinking water for 8 hours / day; 11-14 days old, microbial metabolites for prevention 0.1% for 8 hours / day; 16-20 days old, probiotics for prevention 0.1% for 8 hours / day; 23-26 days old, microbial metabolites for prevention 0.1% for 8 hours / day; if there is death or abnormality, add 1% drinking water to feed complex microbial metabolites for 4 days, 24 hours a day, and then add duck-derived complex probiotics for prevention after stabilization.

[0037] 4. Dung sterilization

[0038] Add breeding dung and high-temperature fermentation bacteria to the litter during duck breeding for high-temperature harmless fermentation treatment. Specifically as follows:

[0039] Apply nano-membrane composting fermentation equipment to the litter during duck breeding for high-temperature harmless fermentation treatment. Mix the previous batch of duck breeding litter, dung, and auxiliary materials (new litter) in a mass ratio of 6:3:1, and add 0.01% high-temperature litter bacteria for fermentation. First, use intermittent ventilation until the temperature rises above 70°C, and continue for 5 days, then switch to continuous ventilation mode.

[0040] III. Results and analysis

[0041] 1. Harmless treatment of litter and dung (previous batch) before breeding

[0042] Apply intelligent molecular membrane fermentation system, add breeding dung and fermentation bacteria to the mixed material for high-temperature harmless fermentation treatment of the litter during duck breeding for 10 days, during which the high temperature above 70°C lasts for 5 days (to ensure harmless treatment of the litter), then increase the ventilation volume of the fan (20 cubic meters / minute) to ensure that the litter is dried in a short time for reuse of the next batch.

[0043] Table 2. Killing effect of different fermentation temperatures on harmful pathogens

[0044]

[0045]

[0046] Table 3. Relative abundance of harmful bacteria and viruses in litter after harmless treatment at different fermentation temperatures detected by metagenome and metavirome

[0047]

[0048]

[0049] From table 2, table 3 can be seen, above 65 DEG C fermentation temperature, the relative abundance of harmful bacteria and viruses in the litter is reduced to 0, the harmless treatment effect is obvious.

[0050] Table 4 different litter high temperature fermentation time on the killing effect of harmful pathogens

[0051]

[0052] From table 4 can be seen, the litter high temperature fermentation 10 days, the treatment effect of harmful bacteria and viruses in the litter is obvious.

[0053] Table 5 the effect of litter treatment, feed fermentation and the addition of beneficial microorganisms and metabolites on duck breeding

[0054]

[0055]

[0056] From table 5 can be seen, using the method of the application, can improve the immunity of duck, improve the absorption of feed, avoid the use of antibiotics, and truly realize the non antibiotic breeding.

[0057] 2, non antibiotic technology in breeding process

[0058] (1) drinking water prevention

[0059] 1-5 day old, add duck source complex probiotic drinking water prevention, 8 hours drinking / D, last for 5 days; 11-14 day old, feed complex beneficial microorganism source metabolite prevention, 8h per day; 16-20 day old duck source complex probiotic prevention 8h / D; 23-26 day old complex beneficial microorganism source metabolite prevention 8h / D; if there is death abnormality, feed micro complex beneficial microorganism source metabolite for 4 days, 24h per day, after stabilization, add duck source complex probiotic prevention 8h / D, last for 3 days. Magpie duck in 40-45 day old, feed 5D complex beneficial microorganism source metabolite prevention 8h / D.

[0060] Table 6 the effect of adding beneficial microorganisms and metabolites on duck breeding

[0061]

[0062] (2) fermented feed use

[0063] The beneficial complex microbial agent separated from duck source intestinal tract is added into the full price feed of duck to ferment and prepare the fermented feed of duck. The fermented feed is added throughout.

[0064] Table 7 the effect of fermented feed and beneficial bacteria on feed to meat ratio, death rate and antibiotic use

[0065]

[0066] Example 1 Raising and management of Muscovy duck

[0067] The preparation of duck-derived intestinal tract beneficial complex microbial agent is as follows:

[0068] 45-day-old healthy Muscovy duck wings were collected for venous blood and feces, and PCR detection was used to exclude related viruses and bacterial pathogens of Muscovy duck. Then the healthy Muscovy duck was sacrificed by heart bloodletting. Then the contents of the muscular stomach, duodenum, ileum, jejunum, cecum and rectum of the Muscovy duck were aseptically collected, added with BHI liquid medium, proliferated at 33°C for 2h, and then plated on BHI plates and proliferated in an aerobic and anaerobic incubator for 48h. Single colonies were picked and plated in BHI broth for aerobic and anaerobic culture for 24h, and then DNA was extracted for in vitro amplification by PCR, and finally sent to a sequencing company for sequencing identification. The identified beneficial bacteria were further subjected to biochemical and related toxicity tests to determine their probiotic properties.

[0069] A total of 100 strains of beneficial bacteria were isolated, including 30 strains of Bacillus distributed in various intestinal segments, 15 strains of Lactobacillus mainly distributed in the stomach and duodenum, 30 strains of Enterococcus distributed in various intestinal segments except the muscular stomach, 12 strains of Bacteroides distributed in various intestinal segments except the muscular stomach, 13 strains of yeast distributed in various intestinal segments except the muscular stomach, and 5 strains of Bifidobacterium distributed in the cecum.

[0070] The above-mentioned bacteria were proliferated in their respective specific culture media to make the viable count of each type of bacteria reach 1.0x10 8 CFU / mL. Then they were mixed in a weight ratio of 2:2:3:1:1:1, inoculated into a mixed culture medium of peptone, yeast extract, glucose and sodium chloride (3:2:3:2) at 2%, and then mixed and fermented at 33°C for 72h.

[0071] (I) Raising during the brooding period (1-3 weeks old)

[0072] 1. Drinking water and feeding. The drinking water and feeding of the hatched Muscovy duckling should be controlled to drink water first and then feed. The feeding time is generally 12-24 hours after hatching, or when 1 / 3 of the ducklings have foraging behavior. Appropriate amount of glucose is added to the drinking water, and then duck-derived complex probiotics are added to the drinking water. From 1-5 days old, duck-derived complex probiotics are used for drinking water prevention; from 11-14 days old, complex beneficial microbial metabolites are used for drinking water prevention; from 16-20 days old, duck-derived complex probiotics are used for drinking water prevention; from 23-26 days old, complex beneficial microbial metabolites are used for drinking water prevention; the above cycle is used for drinking water, and drinking water is provided for 8 hours a day.

[0073] 2. Feed nutrition and formula. Metabolic energy 12.13-12.55 mega joule, crude protein 19.5-22%, calcium 1-1.2%, phosphorus 0.35-0.45% per kilogram of feed. Feed reference formula (%): corn 45, bran 5, fine chaff 7, soybean meal 17, imported fish meal 6, and appropriate amount of minerals and vitamins. Then, the beneficial complex microbial agent separated from the intestinal tract of the duck is added to the complete feed of the duck for fermentation, and the temperature and humidity are referred to the comprehensive pH and temperature of the microbial flora (generally 33°C, humidity 18%, fermentation time 3 days) for solid feed fermentation production to prepare the fermented feed of the duck.

[0074] (ii) Middle brooding period (4-7 weeks old) feeding

[0075] 1. Feeding. The Muscovy duck grows fastest in this stage, the feed intake increases greatly, the digestive capacity is enhanced, and the coarse feed is tolerated. The provided feed can be reduced in quality and increased in quantity, supplemented with green feed and animal feed to meet the Muscovy duck's demand for feed quantity and quality to the maximum extent. However, the Muscovy duck is prone to overeating, and the feeding amount should be properly controlled. The feed change should have a one-week transition period. Metabolic energy 11.7-12.55 mega joule, crude protein 18-19% per kilogram of feed. Feed reference formula (%): corn 50, No. 4 flour 14, bran 8, fine chaff 10, soybean meal 14, fish meal 4, and appropriate amount of minerals and vitamins. The beneficial complex microbial agent separated from the intestinal tract of the duck is added to the complete feed of the duck for fermentation, and the temperature and humidity are referred to the comprehensive pH and temperature of the microbial flora (generally 33°C, humidity 18%, fermentation time 3 days) for solid feed fermentation production to prepare the fermented feed of the duck.

[0076] (iii) Fattening period (8 weeks old to market) feeding

[0077] The proportion of energy feed of the Muscovy duck should be appropriately increased in the fattening stage, and the metabolic energy per kilogram of feed should not be less than 13 mega joule, and the crude protein should be 14-15%. Feed reference formula (%): corn 60, No. 4 flour 10, bran 8, fine chaff 8, soybean meal 12, fish meal 2, and appropriate amount of minerals and vitamins. The beneficial complex microbial agent separated from the intestinal tract of the duck is added to the complete feed of the duck for fermentation, and the temperature and humidity are referred to the comprehensive pH and temperature of the microbial flora (generally 33°C, humidity 18%, fermentation time 3 days) for solid feed fermentation production to prepare the fermented feed of the duck.

[0078] Although the specific embodiments of the present application are described above, those skilled in the art should understand that the specific examples described are only illustrative, and are not intended to limit the scope of the present application, and equivalent modifications and variations made by those skilled in the art in accordance with the spirit of the present application should be covered within the scope of the claims of the present application.

Claims

1. A method for rearing ducks without resistance, characterized by: It comprises the following steps: (1) Preparation of fermented feed for ducks Ferment duck complete feed with duck-derived compound probiotics, and feed the prepared fermented feed as 100% of the daily ration; (2) Adding beneficial bacteria to drinking water A. Drinking water with added duck-derived compound probiotics; B. Drinking water with added compound beneficial microbial metabolites; The drinking method is as follows: From 1-5 days old, drink duck-derived compound probiotic water for prevention; from 11-14 days old, drink compound beneficial microbial metabolite water for prevention; from 16-20 days old, drink duck-derived compound probiotic water for prevention; from 23-26 days old, drink compound beneficial microbial metabolite water for prevention; the above cycle is repeated, and drinking water is provided for 8 hours a day; (3) Bedding sterilization Add breeding feces and high-temperature fermentation-resistant bacteria to conduct high-temperature harmless fermentation treatment on the bedding during duck breeding, and the high temperature above 70℃ lasts for 5 days; The preparation method of the duck-derived compound probiotics is as follows: take the gastrointestinal contents of healthy ducks, increase bacteria in BHI liquid medium, then plate on BHI agar plate, proliferate overnight at 30-35℃, then pick single colonies to BHI liquid medium for proliferation, finally extract strain DNA, PCR amplification, sequencing identification, mix the identified beneficial bacteria for fermentation to prepare duck-derived compound probiotics; The beneficial bacteria are proliferated and mixed, then inoculated into culture medium at an inoculation ratio of 2% for 33℃ mixed fermentation for 72h, and the culture medium comprises the following raw materials in a weight ratio: 3g / L of proteose peptone, 2g / L of yeast extract, 3g / L of glucose, 2g / L of sodium chloride, and prepared duck-derived compound probiotics; The complex beneficial microorganisms in the complex beneficial microorganism metabolic product include Lactobacillus brevis ZG-7, Lactobacillus plantarum, Bacillus, Saccharomycetes and Bifidobacterium, wherein the Lactobacillus brevis ZG-7 is Lactobacillus brevis (Lactobacillus brevis) Levilactobacillus brevis ) ZG-7, which has been preserved in the Guangdong Microbial Culture Collection Center (GDMCC) on June 14, 2023, with a preservation number of GDMCC No. 63562, and the rest are common beneficial bacteria; The preparation method of the compound beneficial microbial metabolites is as follows: mix Lactobacillus brevis ZG-7, Lactobacillus plantarum, Bacillus, yeast, and Bifidobacterium at a weight ratio of 3:2:2:2:1 for mixed culture, proliferate overnight at 30-35℃, and the culture medium comprises the following raw materials in a weight ratio: 8g / L of proteose peptone, 5g / L of yeast extract, 5g / L of glucose, and 2g / L of sodium chloride to obtain the metabolites.

2. The method of rearing ducks without resistance according to claim 1, characterized by: In step (3), the bedding sterilization is to mix the duck breeding bedding, feces, and auxiliary materials at a mass ratio of 6:3:1, and add 0.01% of high-temperature fermentation-resistant bacteria for fermentation.

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