Composition, feed and use for reducing odorous substances in poultry

Abstract

The present invention belongs to the field of biotechnology, and discloses a composition for reducing the emission of odorous substances from poultry, comprising rutin or mulberry leaf extract, Clostridium butyricum, and Bacillus subtilis; the mass ratio of rutin or mulberry leaf extract, Clostridium butyricum, and Bacillus subtilis is 1-3:0.5-1.5:1; the specification of Clostridium butyricum is 1 billion CFU / g, and the specification of Bacillus subtilis is 1 billion CFU / g. The composition utilizes the flavonoid additive rutin or mulberry leaf extract, Bacillus subtilis, and Clostridium butyricum in combination, which can significantly improve the ability of Bacillus subtilis and Clostridium butyricum to inhibit odor. At the same time, the present invention also discloses the application and feed of the composition.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to a composition, feed and use for reducing the emission of odorous substances from poultry. Background Art

[0002] my country is the second largest producer and consumer of broilers in the world. The healthy and sustainable development of broiler farming is directly related to the national economy and people's livelihood. However, while the broiler farming industry is developing rapidly, it is also facing severe challenges. The odor substances in chicken manure are mainly produced by the anaerobic decomposition of undigested nutrients by microorganisms in the feces. They have complex components and diverse types. According to their chemical composition, they are mainly divided into four categories: ammonia and volatile biogenic amines, volatile sulfur compounds, volatile fatty acids, phenols and indole compounds. The production of these odor substances can not only reduce the production performance and product quality of broilers, but also endanger the health of host animals and surrounding residents. By optimizing feeding conditions, scientifically formulating feeds and adding some functional substances, the intestinal health of animals can be improved to a certain extent, the apparent digestibility of nutrients can be increased, the fermentation pattern of intestinal microorganisms can be changed, and the content of odor compounds in animals and feces can be reduced. This is of great significance for promoting the healthy and sustainable development of broiler farming and environmentally friendly animal husbandry production.

[0003] Adding one or more exogenous additives to the diet, such as probiotics, acidifiers, plant extracts, etc., can affect the intestinal microecological structure and thereby reduce the production of odorous substances.

[0004] CN201710683939.6 discloses a feed additive for improving the quality of discarded laying hen meat, and its preparation method and application. The feed additive consists of mulberry leaf powder, green tea powder, betaine, proanthocyanidins and vitamin C, which significantly reduces the odor and ammonia concentration of the chicken house; the mass percentage content of total flavonoids in the mulberry leaf powder is ≥1%.

[0005] The manual further states: The combination of mulberry leaf powder, green tea powder and betaine is more effective in improving the meat quality of eliminated laying hens than using them alone or in combination, and the amount of each component added can be reduced. However, some active components in mulberry leaf powder and green tea powder are easily oxidized, resulting in reduced efficacy and increased addition. When used in combination with proanthocyanidins and vitamin C, the oxidation of active components in mulberry leaf powder and green tea powder can be effectively prevented, and the action ability of mulberry leaf powder, green tea powder and betaine can be improved. At the same time, the deterioration of the meat quality of laying hens due to oxidation can be prevented. The combination of the five components has shown a good effect in improving the meat quality in the production of eliminated laying hens, especially significantly improving the taste and flavor of chicken, and significantly reducing the mortality rate of eliminated laying hens and the odor and ammonia concentration in the chicken house.

[0006] It can be seen that mulberry leaf powder, as a flavonoid active ingredient, can only exhibit corresponding properties in the presence of proanthocyanidins and vitamin C.

[0007] CN202310516234.0 discloses a Clostridium butyricum composite bacterial agent and its application, which points out that the composite bacterial agent includes 55-90 parts of Clostridium butyricum bacterial agent, 6-25 parts of polyglutamic acid, 1-10 parts of Jerusalem artichoke extract, and 3-10 parts of bile acid;

[0008] CN201410843491.6 discloses a Bacillus subtilis and a biological deodorant containing the strain. On the one hand, the strain can effectively reduce the release of malodorous gases such as ammonia and hydrogen sulfide in livestock and poultry manure, thereby reducing the generation of malodorous gases from the source. The ammonia concentration released by chicken manure on the fifth day was only 6.7% of that of the control group, with a significant effect. On the other hand, the strain can also quickly degrade the existing ammonia and hydrogen sulfide in the chicken house. After spraying for 10 minutes, the ammonia concentration in the chicken house was reduced by 95.7%, and the hydrogen sulfide concentration was reduced by 94.7%.

[0009] The applicant previously filed a patent application CN201610115586.5, which discloses a feed additive premix for reducing the odor of broiler feces and its application. The solution uses Bacillus licheniformis, Bacillus subtilis, Clostridium butyricum, Enterococcus faecium, Lactobacillus reuteri, yucca extract, Lauraceae extract, fructooligosaccharides, and rice protein peptides as feed additives, which can effectively reduce the emission of ammonia, hydrogen sulfide and other odorous gases by broilers, improve the growth environment of chickens, and reduce the infection rate and mortality rate of respiratory and intestinal diseases in broilers.

[0010] It can be confirmed without a doubt that Bacillus subtilis and Clostridium butyricum have the function of promoting intestinal health of poultry, reducing odor emissions, and inhibiting the generation of odor from chicken house feces.

[0011] The technical problem to be solved by this scheme is: how to further improve the deodorizing ability of relevant bacteria with deodorizing function. Summary of the invention

[0012] The object of the present invention is to provide a composition for reducing the emission of odorous substances from poultry. The composition utilizes a flavonoid additive rutin or a mulberry leaf extract, Bacillus subtilis, and Clostridium butyricum in combination, and can significantly improve the ability of Bacillus subtilis and Clostridium butyricum to inhibit odor.

[0013] At the same time, the invention also discloses the application of the composition and feed;

[0014] To achieve the above object, the present invention provides the following technical solution: a composition for reducing the emission of odorous substances from poultry, comprising rutin or mulberry leaf extract, Clostridium butyricum, and Bacillus subtilis;

[0015] The mass ratio of rutin or mulberry leaf extract, Clostridium butyricum, and Bacillus subtilis is 1-3:0.5-1.5:1;

[0016] The specification of Clostridium butyricum is 1 billion CFU / g, and the specification of Bacillus subtilis is 1 billion CFU / g.

[0017] In the above-mentioned composition for reducing the emission of odor substances from poultry, the mass ratio of the rutin or mulberry leaf extract, Clostridium butyricum, and Bacillus subtilis is 2:1:1.

[0018] Meanwhile, the present invention also discloses a poultry feed, which comprises a basic feed and a composition accounting for 0.02wt% to 0.08wt% of the weight of the basic feed, and the composition is as shown above.

[0019] In the above-mentioned poultry feed, the basic feed is a corn-soybean meal type basic feed.

[0020] Finally, the present invention also discloses the use of the composition as a feed additive.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] (1) Adding a functional package rich in 0.02% rutin or mulberry leaf extract, 0.01% Bacillus subtilis and 0.01% Bacillus subtilis to the diet of Qingyuan Ma chicken has the effect of improving its growth performance;

[0023] (2) Adding a functional package rich in 0.02% rutin or mulberry leaf extract, 0.01% Bacillus subtilis, and 0.01% Bacillus subtilis to the diet of Qingyuan Ma chickens can improve their nutrient digestibility;

[0024] (3) Adding a functional package rich in 0.02% rutin or mulberry leaf extract, 0.01% Bacillus subtilis, and 0.01% Bacillus subtilis to the diet of Qingyuan Ma chicken can reduce the emission of odorous substances such as p-cresol, indole, ammonia nitrogen, putrescine, and cadaverine from the source, and purify the breeding environment. DETAILED DESCRIPTION

[0025] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. 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.

[0026] 1. Materials and Methods

[0027] The additive rutin used in the experiment was provided by Shanghai Aladdin Technology Co., Ltd. Rutin (purity ≥ 95%) is a flavonoid compound, and the addition amount is 0.02%; Clostridium butyricum and Bacillus subtilis were provided by Jiangsu Yuanshan Biological Company, the specification of Clostridium butyricum was 1 billion CFU / g, and the addition amount was 0.01%; the specification of Bacillus subtilis was 10 billion CFU / g, and the addition amount was 0.01%. The basic diet of the experiment was corn-soybean meal type, and its nutrient content is shown in Table 1.

[0028] Table 1 Basic diet composition and nutritional level

[0029]

[0030]

[0031] Note: 1. Each kg of feed provides: Vitamin A, 3600 IU; Vitamin D, 1650 IU; Vitamin E, 16.5 mg; Vitamin K3, 1.65 mg; Vitamin B1, 1.5 mg; Vitamin B2, 4.5 mg; Vitamin B6, 3 mg; Niacin, 30 mg; Pantothenic acid, 9 mg; Folic acid, 0.75 mg; Vitamin B12, 0.015 mg; Biotin, 0.075 mg; Copper (CuSO4·5H2O),

[0032] 7.2mg; Fe(FeSO4·H2O), 72mg; Zn(ZnSO4·H2O), 66mg; Manganese (MnSO4·H2O), 78mg; I(CI), 0.42mg; See (Na2SeO3), 0.29mg.

[0033] 2. Use the calculated values ​​of feed composition and nutritional value provided by the China Feed Database (2020), except for crude protein.

[0034] 2. Experimental animals and breeding methods

[0035] The experiment selected 1200 Qingyuan Ma chickens aged 90 days and randomly divided them into 10 groups according to body weight, with 6 replicates in each group and 20 chickens in each replicate. The control group was fed a basal diet (control group), and the experimental groups were fed a basal diet + 0.02% rutin group (experimental group 1), a basal diet + 0.02% mulberry leaf extract group (experimental group 2), a basal diet + 0.01% Clostridium butyricum group (experimental group 3), a basal diet + 0.01% Bacillus subtilis group (experimental group 4), a basal diet + 0.02% rutin + 0.01% Clostridium butyricum group (experimental group 5), a basal diet + 0.02% rutin + 0.01% Bacillus subtilis group (experimental group 6), basic diet + 0.02% rutin + 0.01% Bacillus subtilis + 0.01% Clostridium butyricum (experimental group 7), basic diet + 0.02% mulberry leaf extract + 0.01% Bacillus subtilis + 0.01% Clostridium butyricum (experimental group 8), basic diet + 0.01% Bacillus subtilis + 0.01% Clostridium butyricum (experimental group 9). The pre-trial period of the experiment was 5 days, and the formal experiment was 30 days. During the experiment, all Qingyuan Ma chickens were raised in the same chicken house, raised at room temperature, and the relative humidity was maintained at 55-65%. Feeding was carried out at 8:00 every morning, and the leftovers were weighed and recorded, and free access to food and drinking water was allowed. Clean the chicken house, clean up the feces, disinfect and ventilate the chicken house according to weather conditions, and maintain a good sanitary environment in the chicken house.

[0036] One day before the end of the experiment, fresh feces samples of each replicate were collected after feeding was completed. 50-100 g of feces samples were collected from each replicate and placed in a sealed bag. After removing impurities and mixing them thoroughly, they were stored at -80°C for testing of relevant indicators.

[0037] 3. Calculation method

[0038] 3.1 Calculation of growth performance

[0039] All groups were weighed before feeding in the morning at the beginning and end of the experiment, and the initial and final weights were calculated. The remaining amount and feed intake were recorded before and after feeding every day during the experiment. The average daily feed intake (ADFI), average daily weight gain (ADG) and feed-to-gain ratio (F / G) were calculated.

[0040] ADG (g / d) = (final weight - initial weight) / number of test days

[0041] ADFI (g / d) = total material consumption / test days

[0042] FCR=ADFI / ADG

[0043] 3.2 Apparent digestibility of nutrients

[0044] At the end of the feeding experiment, two experimental chickens with a body weight close to the average weight were selected from each replicate, and six replicates were selected for each treatment. They were transferred to metabolic cages, and fecal samples were collected at 8 am every day for three consecutive days to carry out metabolic experiments. The collected fecal samples were placed in a 65℃ oven for drying. After complete drying, they were placed in the air for 1 day to regain moisture, weighed, and crushed to determine the content of each nutrient in the fecal samples.

[0045] Nutrient apparent digestibility (%) = (nutrient mass of ingested feed - nutrient mass of feces) / nutrient mass of ingested feed × 100

[0046] 3.3 Number of main microorganisms in fecal samples:

[0047] About 0.2 g of fecal sample was weighed, and the DNA of microorganisms was extracted according to the kit (StoolDNA Purification Kit) of Hangzhou Xinjing Biological Reagent Development Co., Ltd. Then, according to the method of Yu et al., the number of total bacteria, Lactobacillus, Escherichia coli, and Bifidobacterium in the fecal sample was quantified by fluorescence quantitative PCR instrument. The results were expressed as the number of copies of 16S rRNA gene per gram of fecal sample.

[0048] 3.4 Short-chain fatty acids:

[0049] Weigh about 0.5 g of fecal sample, pre-treat it using the method commonly used in this laboratory, and then measure it using gas chromatography.

[0050] 3.5 Determination of nitrogen-containing metabolic compounds

[0051] According to the methods commonly used in our laboratory in the early stage, the content of ammonia nitrogen, biogenic amines, phenols, indoles and other odorous substances in fecal samples was determined by colorimetry and high performance liquid chromatography.

[0052] 4 Test results

[0053] 4.1 Test of growth performance of Qingyuan Silkie chicken

[0054] The growth performance of Qingyuan Ma chicken is shown in Table 2, as follows:

[0055] Table 2 Effects of rutin and probiotics on the growth performance of Qingyuan Ma chicken

[0056]

[0057]

[0058] As shown in Table 2, compared with the control group, experimental group 3, experimental group 4 and experimental group 9, the final weight of broilers in experimental group 7 increased (P<0.05); the average daily weight gain of experimental groups 7 and 8 was also significantly higher than that of the control group, experimental group 1, experimental group 2, experimental group 3, experimental group 4, experimental group 5, experimental group 6 and experimental group 7 (P<0.05), while the feed-to-weight ratio of experimental groups 7 and 8 was significantly lower than that of the control group, experimental group 1, experimental group 2, experimental group 3 and experimental group 4 (P<0.05).

[0059] From the above results we can see that:

[0060] 1. The feed-to-weight ratios of the control group, experimental group 1, and experimental group 2 show little difference among the three groups. Among them, the performance of experimental group 1 is slightly better, indicating that rutin is slightly better than mulberry leaf extract in promoting the growth performance of chickens;

[0061] 2. The feed-to-weight ratios of the control group, experimental group 3, experimental group 4, and experimental group 8 show that there is little difference between the first three. Among them, the performance of experimental group 3 is slightly better, indicating that the use of probiotics alone has little effect on the growth performance of chickens; the performance of experimental group 8 is significantly better than that of experimental groups 3 and 4, indicating that the use of two probiotics significantly promotes the growth of chickens, which may also be due to the increase in the amount of probiotics used;

[0062] 3. The weight-to-weight ratios of the control group, experimental group 3, experimental group 4, experimental group 5, and experimental group 6 show that once rutin is combined with any probiotic, it can produce a very obvious synergistic effect;

[0063] 4. It can be seen from experimental groups 7 and 8 that the synergistic effect of rutin is significantly better than that of mulberry leaf extract.

[0064] 5. By comparing experimental group 9, experimental group 4, experimental group 5 and experimental group 7, it can be found that the combination of rutin and two probiotics produces a very obvious internal synergistic effect.

[0065] 4.2 Test of nutrient digestibility of Qingyuan Silkie chicken

[0066] The nutrient digestibility of Qingyuan Ma chicken is shown in Table 3 as follows:

[0067] Table 3 Effects of rutin and probiotics on nutrient digestibility of Qingyuan chicken

[0068]

[0069] As shown in Table 3, the apparent digestibility of crude protein in experimental groups 7 and 8 increased significantly (P<0.05) compared with the control group, experimental group 1, experimental group 2, experimental group 3, experimental group 4, and experimental group 9. At the same time, the apparent digestibility of digestible energy in experimental group 7 was also significantly higher than that in the control group, experimental group 1, experimental group 2, experimental group 3, and experimental group 4 (P<0.05).

[0070] The general results are the same as the trend in Table 2, with a slight difference that the trend is opposite to that in Table 2 when two probiotics are used alone.

[0071] 4.3 Test of main microorganisms in Qingyuan Ma chicken feces

[0072] The main microorganisms in Qingyuan Ma chicken feces refer to Table 4, as follows:

[0073] Table 4 Effects of rutin and probiotics on the main microorganisms in Qingyuan Ma chicken feces (lg16S rRNA gene copy number)

[0074]

[0075] As shown in Table 4, compared with the control group, experimental group 1, experimental group 2, experimental group 3, experimental group 4 and experimental group 9, the number of Lactobacillus and Bifidobacterium in fecal samples of experimental group 7 and experimental group 8 increased significantly (P<0.05), while the number of Escherichia coli decreased significantly (P<0.05). At the same time, the number of Bifidobacterium in the control group was significantly lower than that in experimental group 4, experimental group 5, experimental group 6 and experimental group 9 (P<0.05).

[0076] 4.4 Test of short-chain fatty acid content in Qingyuan hemp feces

[0077] The content of short-chain fatty acids in Qingyuan Ma chicken feces is shown in Table 5, as follows:

[0078] Table 5 Effects of rutin and probiotics on the content of short-chain fatty acids in Qingyuan Ma chicken feces (μmol / g)

[0079]

[0080]

[0081] As shown in Table 5, compared with the control group, experimental group 1, experimental group 2, experimental group 3 and experimental group 4, the contents of acetate, butyrate and total short-chain fatty acids in experimental groups 7 and 8 were significantly increased (P<0.05), and the contents of ethylbutyrate in experimental groups 7 and 8 were also significantly higher than those in experimental groups 5 and 6 (P<0.05).

[0082] 4.5 Test of short-chain fatty acid content in Qingyuan hemp feces

[0083] The content of short-chain fatty acids in Qingyuan Ma chicken feces is shown in Table 6, as follows:

[0084] Table 6 Effects of rutin and probiotics on the content of main odorous substances in Qingyuan Ma chicken feces

[0085]

[0086]

[0087] As shown in Table 6, compared with the control group, experimental group 1, experimental group 2, experimental group 3 and experimental group 4, the concentrations of major odorous substances in fecal samples of experimental group 7, such as p-cresol, indole, putrescine and cadaverine, were significantly reduced (P<0.05), the skatole concentrations of experimental groups 7 and 8 were also significantly lower than those of the control group, experimental group 1 and experimental group 2 (P<0.05), and the ammonia nitrogen concentrations of experimental groups 7 and 8 were also significantly lower than those of the control group (P<0.05).

[0088] The above results show that adding a functional package rich in 0.02% rutin, 0.01% Bacillus subtilis and 0.01% Bacillus subtilis to the diet of Qingyuan Ma chicken can improve its growth performance and nutrient digestibility, thereby reducing the emission of odorous substances such as p-cresol, indole, ammonia nitrogen, putrescine, cadaverine, etc. from the source, and purifying the breeding environment.

[0089] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be considered exemplary and non-restrictive in all respects, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims be included in the present invention.

Claims

1. A composition for reducing the emission of odorous substances from poultry, characterized in that: It is composed of rutin, Clostridium butyricum, and Bacillus subtilis; The mass ratio of rutin, Clostridium butyricum, and Bacillus subtilis is 2:1:1; The specification of Clostridium butyricum is 1 billion CFU / g, and the specification of Bacillus subtilis is 1 billion CFU / g.

2. A poultry feed, characterized in that: The poultry feed comprises a basic feed and a composition accounting for 0.02wt% to 0.08wt% of the weight of the basic feed, and the composition is as shown in claim 1.

3. The poultry feed according to claim 2, characterized in that: The basic feed is a corn-soybean meal type basic feed.

4. Use of the composition as claimed in claim 1 as a feed additive.

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