A feed additive suitable for danzhou chicken and a preparation method thereof

Abstract

The application provides a feed additive suitable for Danzhou chicken and a preparation method thereof, which comprises a Callicarpa nudiflora complex, a wine lees fermentation extract and an eleocharis tuberose peel; the wine lees fermentation extract is obtained by fermenting wheat flour and egg white powder with sweet wine lees, and then performing enzymatic dissolution with protease and glucanase, and then concentrating and drying; the feed additive suitable for Danzhou chicken is prepared from the Callicarpa nudiflora complex, the wine lees fermentation extract and the eleocharis tuberose peel; the feeding test proves that the Callicarpa nudiflora complex, the wine lees fermentation extract and the eleocharis tuberose peel have a good promoting effect on the production performance of Danzhou chicken, effectively improve the feed conversion rate, and reduce the mortality rate; and the inhibition rates of the Callicarpa nudiflora complex, the wine lees fermentation extract and the eleocharis tuberose peel on six kinds of bacteria, including Salmonella enteritidis, Salmonella typhi, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Proteus mirabilis, are all above 90%.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of poultry breeding, in particular to a feed additive suitable for Danzhou chicken and a preparation method thereof. BACKGROUND

[0002] Danzhou chicken, also known as Danzhou small breed chicken, stone chicken, etc., is a local characteristic germplasm resource group in Danzhou City, Hainan Province, and is a local soil chicken in Danzhou which is obtained through long-term breeding and domestication by local people in Danzhou. With the efforts of local government departments, scientific research units and colleges and universities, the Danzhou chicken germplasm resource preservation work has achieved certain results, and a Danzhou chicken original seed field has been established. With the improvement of living standards, consumers' demand for high-quality local soil chicken varieties and meat quality is increasing, and the research and development and application of feed additives, as well as disease prevention and control technology, are one of the solutions to improve the market competitiveness of high-quality local chickens. According to the No. 194 announcement of the Ministry of Agriculture and Rural Affairs of the People's Republic of China, all growth-promoting drug feed additives except Chinese herbal medicines are prohibited, and at present, Chinese herbal medicines and extracts, enzyme preparations, and antibacterial peptides used as feed additives have become a research hotspot. Studies have shown that plant biotin, postbiotics, and prebiotics as feed additives can help increase the abundance of intestinal microflora and promote the production of beneficial "metabolites" by microflora, thereby improving the growth performance of poultry.

[0003] The prior art lacks the use of fermented mash as a feed additive for the breeding of Danzhou chicken, and the effects of fermented mash on the growth performance and immunity of Danzhou chicken are studied, so as to provide a reference for the application of the combination of fermented mash and naked flower purple pearl compound in the breeding of Danzhou chicken. SUMMARY

[0004] In view of this, the purpose of the present application is to provide a feed additive suitable for Danzhou chicken and a preparation method thereof, to study the effects of fermented mash on the growth performance and immunity of Danzhou chicken, and to provide a reference for the application of the combination of fermented mash and naked flower purple pearl compound in the breeding of Danzhou chicken.

[0005] The technical solution of the present application is as follows:

[0006] A feed additive suitable for Danzhou chicken, comprising a naked flower purple pearl compound, a wine yeast mash fermentation extract, and an elephant ear skin; the wine yeast mash fermentation extract is a wine yeast mash fermentation product obtained from sweet wine yeast fermentation wheat flour and egg white powder, which is subjected to enzymatic dissolution with protease and glucanase, and then concentrated and dried.

[0007] Further, the naked flower purple pearl compound comprises naked flower purple pearl extract powder, lactose, and micro-powder silica gel in a mass ratio of 0.5-1.5:8-9:0.5-0.8.

[0008] Further, the mass ratio of the naked flower of purple gem, the fermented extract of distiller's yeast and the lotus root skin is 2-4:2-4:0.5-1.

[0009] Further, the adding amount of the feed additive is 300-600mg / kg.

[0010] Further, the preparation method of the fermented extract of distiller's yeast comprises the following steps: mixing wheat flour and egg white powder, adding distilled water with the weight ratio of 1:8-12, stirring, adding sweet wine starter with the inoculation amount of 0.15-0.3% after cooking and cooling, culturing, and obtaining the fermented extract of distiller's yeast.

[0011] Further, the culture condition is as follows: the culture temperature is 33-37℃, the culture oscillation frequency is 150-300r / min, and the culture time is 40-55h.

[0012] Further, the preparation method of the fermented extract of distiller's yeast comprises the following steps: taking the fermented extract of distiller's yeast, centrifuging, taking the precipitate, adding distilled water with the weight ratio of 1:7-9, mixing, adding protease and glucanase with the amount of 0.25-0.35%, enzymatic dissolution at 42-48℃ for 36-45h, inactivating the enzyme, concentrating, and drying to obtain the fermented extract of distiller's yeast.

[0013] Further, the mass ratio of the protease and the glucanase is 3-4:1, and the protease is papain.

[0014] The application further provides a preparation method of the feed additive suitable for Danzhou chicken.

[0015] (1) crushing: drying the lotus root skin at 30-45℃ to constant weight, crushing, and obtaining lotus root skin powder;

[0016] (2) compounding: taking naked flower of purple gem extract powder, lactose and micro-powder silica gel according to the ratio, respectively passing through 50-70 meshes, mixing, and obtaining naked flower of purple gem compound;

[0017] (2) extracting: mixing wheat flour and egg white powder, adding distilled water with the weight ratio of 1:8-12, stirring, adding sweet wine starter with the inoculation amount of 0.15-0.3% after cooking and cooling, culturing at 33-37℃ and 150-300r / min for 40-55h to obtain the fermented extract of distiller's yeast; taking the fermented extract of distiller's yeast, centrifuging, taking the precipitate, adding distilled water with the weight ratio of 1:7-9, mixing, adding papain and glucanase with the amount of 0.25-0.35%, enzymatic dissolution at 42-48℃ for 36-45h, inactivating the enzyme, concentrating, and drying to obtain the fermented extract of distiller's yeast; the mass ratio of the papain and the glucanase is 3-4:1;

[0018] (3) Product: the naked flower purple gem complex powder, the powder of the eleocharis tuberosa peel and the fermented extract of the wine starter are mixed according to the formula to obtain a feed additive suitable for Danzhou chicken.

[0019] Compared with the prior art, the present application has the following beneficial effects:

[0020] (1) The present application provides a feed additive suitable for Danzhou chicken, wherein the components of the feed additive are naked flower purple gem complex, fermented extract of wine starter mash and eleocharis tuberosa peel. The feeding test proves that the naked flower purple gem complex, the fermented extract of wine starter mash and the eleocharis tuberosa peel as the feed additive of Danzhou chicken have a good promoting effect on the production performance of Danzhou chicken, effectively improve the feed conversion rate and reduce the mortality rate.

[0021] (2) The naked flower purple gem complex, the fermented extract of wine starter mash and the eleocharis tuberosa peel provided by the present application have a certain influence on the tenderness of the meat of Danzhou chicken as the feed additive on the basis of the daily ration, and the muscle tenderness is better. The dosage of the feed additive is small, and the feed additive has no toxic side effects on Danzhou chicken and is green and environmentally friendly.

[0022] (3) Adding different dosages of the above-mentioned feed additive has different degrees of reducing effect on the mortality rate of Danzhou chicken, especially in the brooding period, which can effectively promote the absorption efficiency of the chicks to the nutrients, thereby laying the material foundation for the chicks, achieving the effect of improving the immunity of the body and effectively reducing the mortality rate.

[0023] (4) The naked flower purple gem complex, the fermented extract of wine starter mash and the eleocharis tuberosa peel provided by the present application have a bacteriostatic rate of more than 90% on six kinds of bacteria, including enteritis salmonella, typhoid salmonella, staphylococcus aureus, escherichia coli, listeria monocytogenes and proteus mirabilis, and have good bacteriostatic effect. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 It is a statistical result graph of the weight gain rate of each treatment group on the Danzhou chicken in the brooding period;

[0025] Figure 2 It is a statistical result graph of the feed conversion rate of each treatment group on the Danzhou chicken in the brooding period;

[0026] Figure 3 It is a statistical result graph of the mortality rate of each treatment group on the Danzhou chicken in the brooding period;

[0027] Figure 4 It is a statistical result graph of the weight gain rate of each treatment group on the Danzhou chicken in the growing period;

[0028] Figure 5 It is a statistical result graph of the feed conversion rate of each treatment group on the Danzhou chicken in the growing period;

[0029] Figure 6Statistical results of mortality of Danzhou chickens in the growing period for each treatment group;

[0030] Figure 7 In-vitro antibacterial results of four formulae of the feed group against Salmonella enteritidis, Salmonella typhi, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Proteus mirabilis. DETAILED DESCRIPTION

[0031] In order to better understand the technical content of the present application, the following specific examples are provided to further illustrate the present application.

[0032] The experimental methods used in the embodiments of the present application are all conventional methods unless otherwise specified.

[0033] The materials, reagents, etc. used in the embodiments of the present application can be obtained from commercial channels unless otherwise specified.

[0034] Example 1 - Preparation of koji mash fermentation extract

[0035] 1. Experimental materials

[0036] Wheat flour, egg white powder, sweet koji (Angel Yeast Co., Ltd.).

[0037] 2. Koji mash fermentation

[0038] Mix the wheat flour and egg white powder, add distilled water with a solid-liquid ratio of 1:10, and stir, then cook for 20 min. After cooling, evenly mix in the sweet koji at an inoculation amount of 0.2%, and culture at 35℃ and 200 r / min for 48 h to obtain the koji mash fermentation product.

[0039] 3. Extraction of koji mash fermentation product

[0040] Centrifuge the koji mash fermentation product at 8000 r / min for 10 min, take the precipitate, add distilled water with a solid-liquid ratio of 1:8, and blow to make a uniform suspension of the koji mash fermentation product. Add papain and dextranase (with a dosage ratio of 3:1), and the enzyme addition amount is 0.3%. Perform enzymatic dissolution at 45℃ for 40 h, then inactivate the enzyme at 90℃ for 15 min. Concentrate and dry to obtain the koji mash fermentation extract.

[0041] Example 2 - Feeding test of feed additives

[0042] 1. Experimental materials

[0043] Feed additives (naked flower purple gem complex, koji mash fermentation extract and water chestnut peel), basic daily ration (Hainan Chengmai Xinhao Farming and Breeding Co., Ltd., specification: Guoxiong 320) and yeast hydrolysate (Guangzhou Jideyi Biological Technology Co., Ltd.), wherein the naked flower purple gem complex is naked flower purple gem extract powder (Hainan Juzhitang Pharmaceutical Co., Ltd.), lactose and micro-powder silica gel.

[0044] 2. Experimental design

[0045] 375 one-day-old Danzhou chicks of the same size, healthy and lively, and close to the weight were randomly divided into 5 groups, with 5 replicates in each group, 15 in each replicate, i.e. 75 per treatment group. Chicks with consistent feather color and close to the weight were selected for the test. Three chicks per replicate and close to the average value in the group were selected for blood sampling and slaughter. The experimental site was the Livestock Research Center of the Institute of Tropical Crop Variety Resources of the Chinese Academy of Tropical Agricultural Sciences in Danzhou, Hainan Province. During the experiment, the Danzhou chicks were free to eat and drink water, and the daily management was the same as the relevant procedures of the Danzhou chicken farm. The test lasted for 48 days. The feeding methods of each group of Danzhou chicks were as follows:

[0046] Feeding group one (control group): feeding basic diet;

[0047] Feeding group two (comparison group): feeding basic diet + 300 mg / kg feed additive (the use amount ratio of naked flower purple pearl compound, yeast hydrolysate and arrowhead skin is 3:3:1);

[0048] Feeding group three: feeding basic diet + 300 mg / kg feed additive (the use amount ratio of naked flower purple pearl compound, wine yeast extract and arrowhead skin is 3:3:1);

[0049] Feeding group four: feeding basic diet + 300 mg / kg feed additive (the use amount ratio of naked flower purple pearl compound, wine yeast extract and arrowhead skin is 3:0.5:1);

[0050] Feeding group five: feeding basic diet + 600 mg / kg feed additive (the use amount ratio of naked flower purple pearl compound, wine yeast extract and arrowhead skin is 3:3:1);

[0051] Among them, the naked flower purple pearl compound is: naked flower purple pearl extract powder, lactose and micro-powder silica gel are respectively passed through a 60-mesh sieve in a ratio of 1:8.5:0.5, and then uniformly mixed to obtain the naked flower purple pearl compound; the arrowhead skin is dried in a 40°C drying oven until the weight is constant, and then pulverized to obtain arrowhead skin powder

[0052] The naked flower purple pearl compound, arrowhead skin powder and wine yeast extract are uniformly mixed in a mixer according to the ratio to obtain the feed additive.

[0053] 3. Determination index

[0054] (1) Growth performance determination

[0055] Average body weight (g) = total weight (g) / total number of chickens (number);

[0056] Average weight gain per chicken = final body weight (g) - initial body weight (g);

[0057] Average feed intake per chicken = total feed intake (g) / number of chickens (chickens) ;

[0058] Feed conversion rate (%) = average weight gain per chicken (g) / average feed intake per chicken (g) x 100%;

[0059] Mortality rate (%) = number of dead chickens / total number of chickens x 100%.

[0060] (2) Determination of organ index

[0061] After the end of the breeding (2022.9.26), 2 chickens were randomly selected from each group and each repetition, and after being killed, the heart, liver, spleen, kidney, thymus and bursa of Fabricius were removed, the excess tissue was removed, the filter paper was wiped dry, and the weight was recorded. The ratio of internal organ weight to last weight (2022.9.27) body weight was calculated, and the organ index = internal organ weight (mg) / fasting body weight (g).

[0062] (3) Determination of meat quality traits

[0063] Take chicken breast meat about 6 cm x 3 cm x 3 cm in size, remove tendons, fat and other substances, set the water bath to 80℃, and when the center temperature of the chicken breast meat reaches 70℃, take out the meat and cool it. After taking the meat sample hole, the C-LM3B type muscle tenderness instrument was used to measure the shear force of the sample.

[0064] 4. Feeding results of feed additives

[0065] (1) Feeding results of feed additives on Danzhou chickens during the brooding period

[0066] Table 1 Statistical results of feed conversion rate and mortality rate of Danzhou chickens during the brooding period in each group

[0067]

[0068]

[0069] From Table 1, the feed conversion rates of each treatment group were: the control group was 41.7%, the feeding group two was 44.94%, and the different amounts and ratios of feed additives were 45.20%, 42.01% and 42.88%, respectively. The weight gain rates of each treatment group were: the control group was 761.80%, the comparison group was 774.45%, and the different amounts and ratios of feed additives were 829.02%, 742.14% and 766.74%, respectively.

[0070] From the above, the addition of naked flower purple compound, wine koji fermentation extract and water chestnut peel as the feed additive of Dangzhou chicken, has no toxic side effects on Dangzhou chicken, on the contrary, the addition of different doses of the above feed additives has different degrees of reducing effect on the mortality of Dangzhou chicken, and effectively improves the feed conversion rate, from the mortality, different amount, ratio of feed additives are lower than 3%, which shows that naked flower purple compound, wine koji fermentation extract and water chestnut peel as the feed additive of Dangzhou chicken, can effectively promote the absorption efficiency of nutrients of chicks during the brooding period, and then lay the material basis of chicks during the brooding period, achieve the effect of improving the immunity of the body and effectively reducing the mortality.

[0071] In summary, the addition of naked flower purple compound, wine koji fermentation extract and water chestnut peel as the feed additive of Dangzhou chicken, has good promoting effect on the production performance of Dangzhou chicken during the brooding period, and significantly reduces the mortality during the brooding period.

[0072] (2) The results of feed additives on the feeding of Dangzhou chicken during the growing period

[0073] Table 2 The statistical results of feed conversion rate and mortality of each group of Dangzhou chicken during the growing period

[0074] Group Control group Feeding group two Feeding group three Feeding group four Feeding group five Weight gain 399.05±16.43 391.18±15.56 453.15±20.25 404.59±12.88 413.84±5.30 Weight gain rate (%) 202.03±19.45 171.45±10.18 213.00±16.26 180.01±12.00 184.51±7.44 Feed conversion rate (%) 25.79±3.22 25.28±1.43 27.10±1.91 25.18±1.08 25.53±0.82 Mortality rate (%) 3.33±6.67 3.45±3.99 0.00 1.48±2.95 3.45±3.99

[0075] From table 2, the feed conversion rate of each treatment group is: the control group is 25.79%, the feeding group two is 25.28%, the different amount and ratio of feed additives are 27.1%, 25.18% and 25.53% respectively, the weight gain rate of each treatment group is: the control group is 202.03%, the comparison group is 171.45%, the different amount and ratio of feed additives are 213.00%, 180.01% and 184.51% respectively, from the results, only feeding group three can significantly improve the weight gain rate during the growing period, the rest of the groups have no significant effect on the weight gain rate, and each group has no significant effect on the feed conversion rate and mortality, in the case of no difference in feed conversion rate, feeding group three can increase the feed intake of Dangzhou chicken during the growing period, it can be seen that the addition of naked flower purple compound, wine koji fermentation extract and water chestnut peel as the feed additive of Dangzhou chicken, has good promoting effect on the production performance of Dangzhou chicken during the growing period, and further improves the weight gain rate of Dangzhou chicken during the growing period.

[0076] (3) The results of feed additives on the feeding of meat quality

[0077] Table 3 The statistical results of shear force of each group of Dangzhou chicken

[0078] Group Shear force (N) Control group 21.56 Group three 18.72

[0079] As seen from Table 3, after 84 days of cultivation, the shear force of Danzhou chickens in Group Three is lower than that of the control group, and the addition of the naked flower purple gem complex, the extract of fermented mash with distiller's yeast and the feed additive of the eleocharis tuberosa peel has certain influence on the meat tenderness of Danzhou chickens, and the muscle tenderness is better and the meat is more delicate.

[0080] Example 3 - Test of antibacterial effect in vitro

[0081] Preferably, the formula of Group Three is dissolved in a 20% volume concentration of ethanol solution to make the concentration of the formula 0.2 g / mL. The formula is ultrasonically treated for 1 h, and then centrifuged at 10,000 rpm for 10 min at 25°C. The supernatant is taken for standby and determination of the antibacterial rate. Single colonies of Salmonella enteritidis, Salmonella typhi, Staphyloccocus aureus, Escherichia coli, Listeria monocytogenes and Proteus mirabilis are taken into 50 μL of liquid culture medium, and then placed in a 37°C incubator for 12 h. The number of viable colonies is counted under a microscope, and then diluted to 10 x 7 cfu / mL for standby. In a sterile operation table, 0.1 μL of the above bacterial solution is taken and added into 5 mL of the test solution of Group Three and the control solution of 20% volume concentration of ethanol solution. Then, the mixture is placed in a culture incubator for 10 min, which is called the action time. Then, 0.5 mL of each solution after the action is taken and added into 5 mL of PBS solution, and then placed in a culture incubator for 10 min, which is called the termination reaction. Each solution after the termination reaction is diluted with PBS at a ratio of 1:5, and then placed in a culture incubator for 10 min. 0.5 mL of the above diluted solution is taken and added into a culture dish, and then 15 mL of nutrient agar culture medium cooled to 40-45°C is poured into the culture dish for pouring. The culture dish is rotated to mix the solution thoroughly. After the agar is completely solidified, the culture dish is turned over and placed in a culture incubator for 24 h. Then, the number of viable colonies is counted to calculate the antibacterial rate. The antibacterial rate calculation formula is: (control solution - test solution) / control solution x 100%.

[0082] Table 4 - Antibacterial results of Group Three on six kinds of bacteria

[0083] Serial number Bacteria Bacteriostatic rate (%) 1 Salmonella enteritidis 95.8 2 Salmonella typhi 94.2 3 Staphylococcus aureus 91.6 4 Escherichia coli 97.3 5 Listeria monocytogenes 96.1 6 Proteus mirabilis 95.4

[0084] According to the above results, the antibacterial rate of Group Three of the present application on Salmonella enteritidis, Salmonella typhi, Staphyloccocus aureus, Escherichia coli, Listeria monocytogenes and Proteus mirabilis reaches more than 90%.

[0085] The above merely provides the preferred embodiment of the present application, and is not used to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A feed additive suitable for Danzhou chicken, characterized in that, The feed additive comprises the naked flower purple gem complex, the wine starter mash fermentation extract, and the water chestnut peel, and the mass ratio of the naked flower purple gem complex, the wine starter mash fermentation extract, and the water chestnut peel is 3:3:

1. The feed additive is added in an amount of 300 mg / kg. The naked flower purple gem complex comprises the naked flower purple gem extract powder, lactose, and micro-powder silica gel, and the mass ratio of the naked flower purple gem extract powder, lactose, and micro-powder silica gel is 0.5-1.5:8-9:0.5-0.

8. The wine starter mash fermentation extract is obtained by fermenting wheat powder and egg white powder with sweet wine starter, enzymatically dissolving by using protease and glucanase, and then concentrating and drying.

2. The feed additive according to claim 1, characterized in that, The preparation method of the wine starter mash fermentation extract comprises the following steps: taking the wine starter mash fermentation product, centrifuging, taking the precipitate, adding distilled water in a solid-liquid weight ratio of 1:7-9, mixing, adding protease and glucanase in an amount of 0.25-0.35%, enzymatically dissolving at 42-48°C for 36-45 hours, inactivating the enzyme, concentrating, and drying to obtain the wine starter mash fermentation extract.

3. The feed additive according to claim 2, characterized in that, The preparation method of the wine starter mash fermentation product comprises the following steps: mixing wheat powder and egg white powder, adding distilled water in a solid-liquid weight ratio of 1:8-12, stirring, adding sweet wine starter in an inoculation amount of 0.15-0.3% after steaming and cooling, and culturing to obtain the wine starter mash fermentation product.

4. The feed additive according to claim 3, characterized in that, The culture conditions are as follows: a culture temperature of 33-37°C, a culture oscillation frequency of 150-300 r / min, and a culture time of 40-55 hours.

5. The feed additive according to claim 2, characterized in that, The mass ratio of the protease and the glucanase is 3-4:1, and the protease is papain.

6. The method for preparing the feed additive for Danzhou chicken according to any one of claims 1-5, characterized in that, The method comprises the following steps: (1) crushing: drying the water chestnut peel at 30-45°C to constant weight, and crushing to obtain water chestnut peel powder; (2) compounding: taking the naked flower purple gem extract powder, lactose, and micro-powder silica gel according to the ratio, respectively passing through a 50-70 mesh sieve, and mixing to obtain the naked flower purple gem complex; (3) extracting: mixing wheat powder and egg white powder, adding distilled water in a solid-liquid weight ratio of 1:8-12, stirring, adding sweet wine starter in an inoculation amount of 0.15-0.3% after steaming and cooling, and culturing at 33-37°C and 150-300 r / min for 40-55 hours to obtain the wine starter mash fermentation product; taking the wine starter mash fermentation product, centrifuging, taking the precipitate, adding distilled water in a solid-liquid weight ratio of 1:7-9, mixing, adding papain and glucanase in an amount of 0.25-0.35%, enzymatically dissolving at 42-48°C for 36-45 hours, inactivating the enzyme, concentrating, and drying to obtain the wine starter mash fermentation extract; The mass ratio of the papain and the glucanase is 3-4:1; (4) finished product: mixing the naked flower purple gem complex powder, the water chestnut peel powder, and the wine starter mash fermentation extract according to the formula to obtain the feed additive suitable for Dangzhou chickens.

Images