CN114158657A - Functional amino acid for improving oxidation resistance and immunity of poultry and application thereof - Google Patents

Abstract

The invention relates to a functional amino acid for improving the oxidation resistance and immunity of poultry and application thereof, wherein the functional amino acid is 5-aminolevulinic acid, the functional amino acid is added into poultry feed, drinking water and medicines in the poultry breeding industry, the daily intake of the poultry is 0.0001-1g by the 5-aminolevulinic acid, and the addition amount of the functional amino acid in the complete feed for 1-42-day-old broiler chickens is 30-100 g/ton in the complete feed for the poultry; the addition amount of the complete feed for meat ducks of 1-28 days old is 40-100 g/ton; the addition amount of the complete feed for laying hens aged over 28 weeks is 30-100 g/ton. The invention discloses that 5-ALA can improve the oxidation resistance and immunity of poultry, and one or more of 5-aminolevulinic acid, 5-aminolevulinic acid derivatives or 5-aminolevulinic acid salts can be added into poultry feed, drinking water and medicines, so that the oxidation resistance and immunity of poultry can be improved after the poultry ingests the 5-ALA, and the health state and production performance of the poultry can be improved.

Description

Functional amino acid for improving oxidation resistance and immunity of poultry and application thereof

Technical Field

The invention belongs to the technical field of biology, and particularly relates to functional amino acid for improving the oxidation resistance and immunity of poultry and application thereof.

Background

Under normal physiological state and life activity, the generation and elimination of free radicals in animal body always keep a dynamic balance to maintain normal function and activity of animal body. When animals eat the oxidized diet or are affected by adverse factors such as stress and the like, a large amount of free radicals are generated in the animals, and if the free radicals exceed the capacity and speed of the organisms for removing the free radicals, the free radicals are accumulated in the organisms, the oxidation-reduction balance state in the organisms is damaged, the organisms of the animals are in an oxidative stress state, the organisms are affected and damaged, and the growth performance of the animals is reduced and the quality of animal products is reduced.

Poultry such as broiler chicken, meat duck, laying hen and the like can generate free radicals and endogenous active substances for resisting free radicals in the growth metabolic process, and the free radicals and the endogenous active substances are in dynamic balance under normal conditions. However, high energy and high protein feeds, oxidized fats, fast growth, high density feeding, cage farming environments, high temperatures, etc. all cause strong stress reactions in poultry such as broiler chickens, meat ducks, laying hens, etc. Biological antioxidant systems in poultry bodies such as rapidly-growing broilers, meat ducks and laying hens are fragile and easy to be stressed by environment, oxidative stress caused by oxidized grease in feed, high temperature in houses and the like causes imbalance of in-vivo antioxidant reaction, free radicals and peroxide are accumulated, harm and influence are caused to body health, growth performance, meat quality after slaughtering and the like of the poultry, and feed reward is further influenced.

5-Aminolevulinic acid (5-ALA) is a naturally occurring functional amino acid in vivo, and is chlorophyllHeme, VB₁₂The essential precursor of synthesis is a substance indispensable to life activities. 5-aminolevulinic acid is a natural compound produced when a citric acid cycle (TCA) circuit operates in mitochondria of higher cells, a nonprotein amino acid widely present in living cells of living organisms such as bacteria, fungi, animals and plants, and a metabolically active physiologically active substance essential for life activities of animals and plants. At present, no report is found about the research of improving the production performance of livestock and poultry by improving the antioxidant capacity and the immune function of the poultry by using 5-ALA.

Disclosure of Invention

Aiming at the defects of the prior art, the invention considers that the biological oxidation resistance in the rapidly growing poultry is low, the immune system is fragile, and the stress reaction can be caused by the oxidation-reduction imbalance of the organism, so that the invention provides the functional amino acid capable of improving the oxidation resistance and the immune function of the poultry, the effective component of the functional amino acid is 5-aminolevulinic acid, and the production performance is improved by improving the oxidation resistance and the immune function of the poultry.

The invention provides a functional amino acid for improving the oxidation resistance and immunity of poultry, wherein the functional amino acid is 5-aminolevulinic acid, and the active ingredient of the 5-aminolevulinic acid is one or more of 5-aminolevulinic acid, 5-aminolevulinic acid derivatives or 5-aminolevulinic acid salts.

The invention also provides application of the function of improving the oxidation resistance and the immunity of poultry based on 5-aminolevulinic acid in the poultry breeding industry, and particularly provides application of the function of improving the oxidation resistance and the immunity of the poultry based on 5-aminolevulinic acid in poultry feed, drinking water and medicine, wherein one or more of 5-aminolevulinic acid, 5-aminolevulinic acid derivatives or 5-aminolevulinic acid salts are added into the poultry feed, drinking water and medicine, and the daily intake of the poultry is 0.0001-1g calculated by 5-aminolevulinic acid.

Further, one or more of 5-aminolevulinic acid, 5-aminolevulinic acid derivatives or 5-aminolevulinic acid salts are added into the complete feed for the domestic fowls, and the addition amount of the complete feed for the 1-42-day-old broiler chickens is 30-100 g/ton based on the 5-aminolevulinic acid; the addition amount of the complete feed for meat ducks of 1-28 days old is 40-100 g/ton; the addition amount of the complete feed for laying hens aged over 28 weeks is 30-100 g/ton.

Furthermore, the additive amount of the complete feed for 1-42 day old broiler chickens is 45-60 g/ton.

Furthermore, the addition amount of the complete feed for meat ducks of 1-28 days old is 70-100 g/ton.

Furthermore, the addition amount of the complete feed for laying hens aged over 28 weeks is 70-100 g/ton.

The invention has the following beneficial effects:

1. the 5-ALA can improve the oxidation resistance and immunity of poultry, and can be added into poultry feed, drinking water and medicines by one or more of 5-aminolevulinic acid, 5-aminolevulinic acid derivatives or 5-aminolevulinic acid salts, so that the oxidation resistance and immunity of poultry are improved after the poultry ingests the 5-ALA, and the health state and the production performance of the poultry are improved.

2. The adding time and the adding amount of the 5-ALA in the daily ration of the broiler chickens, the meat ducks and the laying hens are determined, the optimal selection is given, the 5-ALA is added into the feed of the broiler chickens of 1-42 days old according to the amount of 30-100 g/ton, and the optimal selection is 45-60 g/ton; adding 5-ALA in an amount of 40-100 g/ton into 1-28 days old meat duck feed, and optimally selecting 70-100 g/ton; 5-ALA is added into the feed according to the amount of 30-100 g/ton from the age of 28 weeks, and the optimal addition amount is 70-100 g/ton.

3. According to the use time and the addition amount of the 5-ALA provided by the invention, the feed-meat ratio of broiler chickens and meat ducks and the feed-egg ratio of laying hens can be obviously reduced; the immune organ index of the tested poultry tends to increase, and has certain positive effects on the establishment of the immune function of the poultry organism and the maintenance of the internal environment stability of an immune system; obviously improves the activity of catalase in poultry serum, obviously reduces the content of malondialdehyde serving as a lipid oxidation marker, and improves the oxidation resistance of poultry, thereby improving the production performance of broiler chicken.

4. According to the using time and the adding amount of the 5-ALA provided by the invention, the IgM, IgG and IgA contents in the broiler serum can be obviously improved, the gamma-interferon (IFN-gamma) level in the broiler serum can be obviously improved, the trend of a serum tumor necrosis factor alpha (TNF-alpha) is reduced, and the IL-4 content in the serum is improved to a certain extent. The results show that the addition of 5-ALA in the daily ration can reduce the content of proinflammatory cytokines in the broiler chicken, improve the level of the anti-inflammatory cytokines, activate the innate and acquired immune systems of the body and promote the improvement of the immune level of the body, thereby improving the production performance of the broiler chicken.

5. According to the application time and the addition amount of the 5-ALA provided by the invention, the IgM, IgG and IgA contents in the blood serum of the meat duck can be obviously improved, the secretory expression of TLR4 in the jejunum of the meat duck is reduced, the transitional secretion of proinflammatory factors TNF-alpha and IL-6 in the intestinal tract is reduced, and the improvement of the immune level of an organism is promoted, so that the production performance of the meat duck is improved.

6. According to the use time and the addition amount of the 5-ALA provided by the invention, the contents of IgA, IgG and IgM in the blood serum of the laying hens can be increased, and when the addition amount is 40-100g/t, compared with a group without the 5-ALA, the content of IgA in the blood serum can be obviously increased, the improvement of the body immunity level is promoted, and the production performance of the laying hens is improved.

Drawings

FIG. 1 is a graph showing the effect of 5-ALA on intestinal bleeding point of broiler chickens at 42 days of age

Detailed Description

In order to make the technical solutions better understood by those skilled in the art, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only partial embodiments of the present application, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.

Example 15 feeding experiments of ALA on broiler chickens and its effects on Productivity, serum Biochemical indices and immune function

1.1 test materials

5-aminolevulinic acid phosphate with content of 5%, and 5-aminolevulinic acid (5-ALA) as active ingredient.

1.2 test animal grouping and feeding management

1680 AA broiler chicks of 1 day old were randomly divided into 15 treatment groups, each treatment group was replicated 6 times, and each treatment group was replicated 20 chickens (10 chickens each male and female, mixed rearing). The test is carried out in two stages, wherein the 1-21 days old is the early stage of growth, and the 22-42 days old is the late stage of growth. The chickens are raised in cages by adopting three layers of overlapped type, are fed freely, drink water freely, are artificially illuminated for 24 hours at the age of 1-3 days, and are kept illuminated for 16 hours and dark for 8 hours. The water circulation type central heating system supplies heat, the temperature of 1-3 days old is maintained at 33 ℃, the temperature is reduced by 2 ℃ every week, and the temperature of the henhouse is finally maintained at 24 ℃. Marek's vaccine was inoculated at 1 day of age, and newcastle disease and renal infectious bronchitis combined vaccine was inoculated by nasal drip at 7 days of age. Henhouse sanitation management is performed conventionally.

1.3 test feed

The basic diet composition and nutritional ingredients for this trial are shown in table 1. A total of 5 experimental diets were prepared, each: t1 (basal diet), T2 (basal diet +10g/T of 5-ALA), T3 (basal diet +15g/T of 5-ALA), T4 (basal diet +20g/T of 5-ALA), T5 (basal diet +25g/T of 5-ALA), T6 (basal diet +30g/T of 5-ALA), T7 (basal diet +35g/T of 5-ALA), T8 (basal diet +40g/T of 5-ALA), T9 (basal diet 45g/T of 5-ALA), T10 (basal diet +50g/T of 5-ALA), T11 (basal diet +55g/T of 5-ALA), T12 (basal diet +60g/T of 5-ALA), T13 (basal diet +70g/T of 5-ALA), T14 (basal diet +100g/T of 5-ALA). The test feed is granulated and then fed.

1.4 measurement index and method

1.4.1 growth Performance

During the test period, only the growth and health condition of the test chicken are observed, and the feed intake is repeatedly recorded. The chickens were weighed in duplicate after 8 hours fasting at 21 and 42 days of age, respectively. And calculating the average weight, the average daily gain, the average daily feed intake, the material consumption and weight gain ratio and the death and culling rate of the plants at 1-21 days old, 22-42 days old and 1-42 days old.

TABLE 1 basic diet composition and nutritional ingredients

1) Vitamins and trace elements premix (each 1kg of batch contains effective components): 8000IU of vitamin A, 31000 IU of vitamin D, 20mg of vitamin E, K30.5mg of vitamin, 12.0 mg of vitamin B, 28 mg of vitamin B, 63.5 mg of vitamin B, 120.01mg of vitamin B, 10mg of pantothenic acid, 35mg of nicotinic acid, 0.55mg of folic acid, 0.18mg of biotin, 8mg of copper sulfate, 100mg of ferrous sulfate, 80mg of zinc sulfate, 80mg of manganese sulfate and 72mg of potassium iodide. 6000 IU of vitamin A, 3750 IU of vitamin D, 10mg of vitamin E, K30.5mg of vitamin, 12.0 mg of vitamin B, 25 mg of vitamin B, 63.0 mg of vitamin B, 120.01mg of vitamin B, 10mg of pantothenic acid, 0.55mg of folic acid, 0.15mg of biotin, 6.4mg of copper sulfate, 64mg of ferrous sulfate, 64mg of zinc sulfate, 64mg of manganese sulfate and 0.56mg of potassium iodide. 2) The nutrient levels are calculated values.

1.4.2 measurement of antioxidant index of liver tissue and serum

Liver tissue (5g) from the same side was taken, washed with sterile saline solution to remove blood traces, and immediately stored in liquid nitrogen and then stored at-70 ℃ for subsequent analysis. A 10% liver homogenate was prepared using a phosphate buffer solution at a weight (g) to volume (mL) ratio of 1: 9. Centrifuging at 4 deg.C (3500rpm) for 10min to obtain homogenate supernatant for antioxidant index analysis. The protein concentration of the liver homogenate was determined by the Bradford method. Total antioxidant capacity (T-AOC), Malondialdehyde (MDA) content, total superoxide dismutase (SOD) and Catalase (CAT) activity in serum and tissue homogenate supernatants were determined using a commercial kit purchased from Nanjing institute of bioengineering.

1.4.3 determination of immune organ index and immune-related index

At 42 days of age, broiler chickens with a weight close to the average weight (1 each for the male and female) were repeatedly selected and slaughtered, and the thymus, spleen and bursa of Fabricius of the chickens were collected, and then the blood stain on the organ surface was removed with filter paper and weighed. Immune organ index (%) ═ 100 x immune organ weight (g)/body weight (g).

Serum concentrations of immunoglobulin a (iga), immunoglobulin m (igm), immunoglobulin g (igg), tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), and interleukin-4 (IL-4) were determined using a chicken-specific ELISA kit purchased from shanghai enzyme linked biotechnology limited (china, shanghai).

1.4.4 statistical analysis: all experimental data were analyzed by the one-way variance (ANOVA) program performed by IBM-SPSS 19.0(SPSS, Chicago, IL) statistical software. The duncan assay was used for multiple comparisons. P <0.05 indicates significance of difference.

1.5 results and analysis

1.5.1 Effect of 5-ALA addition to the daily ration on the growth Performance of broiler chickens

TABLE 2 Effect of 5-ALA addition to diets on broiler growth Performance

The stress and oxidation can cause the increase and accumulation of oxygen free radicals in animal bodies, so as to cause damage to intestinal mucosa, cause atrophy and shedding of intestinal epithelial villi, reduce the digestion and absorption functions of the intestinal epithelial villi and further influence the breeding benefits of broiler chickens. The effect of the addition of 5-ALA to the ration on the productivity of broiler chickens is shown in Table 2.

The result shows that 30-100g/t 5-ALA is added into the broiler feed to reduce the feed conversion ratio in the early stage, the later stage and the whole stage to different degrees. The feed conversion ratio (P <0.05) of 1-21-day-old broiler chickens can be remarkably reduced by feeding 5-ALA added with 45-60g/t of daily ration, the feed conversion ratio (P <0.05) of 22-42-day-old broiler chickens can be remarkably reduced by feeding 5-ALA added with 30-100g/t of daily ration, the average daily gain of 1-42-day-old broiler chickens can be remarkably increased by adding 30-100g/t of 5-ALA, and the feed conversion ratio of the whole period (1-42-day-old) of the broiler chickens can be remarkably reduced by adding 30-70 g/t of 5-ALA. In summary, 5-ALA added to feed for 1-42 day old broiler chickens in an amount of 45-60 g/ton is most preferably selected.

1.5.2 Effect of 5-ALA addition to daily ration on slaughter Performance and immune organ index of broiler chickens

The immune organs of the poultry mainly comprise thymus, spleen and bursa of fabricius, and the increase of the mass of the poultry is generally considered to be caused by the division and growth and development of self cells, which can indicate the improvement of the immune function and the immune level of the body. In the case of healthy animals, the higher the immune organ index, the stronger the immune function of the animal. Studies have shown that thymus, spleen, and bursal index can be used to evaluate immune status. Animal test results show that 15-100 g/t5-ALA has no significant influence on broiler immune organ indexes (Table 3), thymus serving as a primary immune organ dominates the development of peripheral immune organs and is the earliest immune organ to occur, and the index of 5-ALA added with 45-70 g/t has a tendency of increasing along with the increasing of the added amount, so that the 5-ALA has a certain positive effect on establishment of the immune function of a broiler organism and maintenance of immune system homeostasis.

TABLE 3 Effect of 5-ALA addition to diets on broiler immune organs

1.5.3 Effect of 5-ALA added into daily ration on oxidation resistance of broiler blood serum and liver tissue

T-AOC is a comprehensive index for measuring the oxidation resistance of organisms, and MDA is an important index for measuring the lipid peroxidation level and the cell damage degree. Within the normal range, the higher the CAT, T-SOD activity, the higher the T-AOC in serum and liver, and the lower the MDA content. In the modern breeding process, high-energy high-protein feed, oxidized grease, rapid growth, high-density breeding, cage culture environment, high temperature and the like can cause broiler chickens to generate strong stress response, organisms generate a large amount of superoxide anions, hydroxyl free radicals and hydrogen peroxide, and if the superoxide anions, the hydroxyl free radicals and the hydrogen peroxide are not removed in time, the excessive accumulation of the superoxide anions, the hydroxyl free radicals and the hydrogen peroxide in blood and liver is caused, the oxidation resistance of serum and liver is reduced, and a large amount of MDA is generated.

As shown in Table 4, compared with the control group, the activity of catalase in the serum of broiler chickens in the test group added with 35-70 g/t5-ALA in daily ration is obviously improved at 21 days of age (CAT, P is 0.05). The CAT activity level of the broiler chickens at the age of 42 days can be improved by adding 5-ALA into the daily ration. The addition of 45-60 g/T5-ALA in the daily ration has the effect of improving the total antioxidant capacity (T-AOC) in the serum of the broiler chickens with the age of 21 days and 42 days, and has no obvious influence (P is more than 0.05) on Malondialdehyde (MDA) and total superoxide dismutase (T-SOD).

TABLE 4 influence of the addition of 5-ALA to the daily ration on the serum antioxidant capacity of broiler chickens

TABLE 5 Effect of 5-ALA addition to diets on broiler liver tissue antioxidant Capacity

As shown in Table 5, the total antioxidant capacity (T-AOC) of liver tissues of 21-day-old broilers is improved with the increase of the addition level of 5-ALA in daily ration, wherein the addition of 45-60g/T of 5-ALA can obviously improve the T-AOC (P <0.05) of the liver tissues of 21-day-old broilers compared with a control group. In addition, 5-ALA is added into the daily ration to promote Catalase Activity (CAT) of liver tissues, and the CAT enzyme activity is remarkably higher than that of a control group after 10-100 g/t5-ALA is added. In addition, the addition of 35 and 100g/t of 5-ALA to the ration significantly reduced the content of Malondialdehyde (MDA), a marker of lipid oxidation, compared to the control group (P < 0.05). In addition, 5-ALA of 45-60g/t is added into the daily ration, so that the activity of superoxide dismutase (SOD) of 42-day-old broiler chickens is obviously improved. Similarly, the addition of 5-ALA to the diet tended to decrease the amount of MDA in liver tissue (P ═ 0.09).

1.5.4 influence of 5-ALA added into diet on immunity of broiler chicken

Immunoglobulins are the major immune molecules of humoral immunity in animals and play an important role in the immune system of the body. Serum immunoglobulin content can reflect changes in the immune function of the body. IgG is mainly secreted by plasma cells of immune organs, is a key immune protein for mediating humoral immunity, and has higher content in serum; IgM is an immune substance which is firstly secreted and released in the humoral immunity process, and can react with in vivo and in vitro antigens to eliminate pathogens. IgA is secreted by plasmablasts in the intestinal tract, is mainly distributed in the mucus layer, is the core of main immunoglobulin for providing immune protection for the intestinal tract and mucosal immunity, is combined with surface antigens of pathogenic microorganisms to protect intestinal epithelial cells from being damaged, and is a main effector of humoral immunity of the broiler chicken. The immunoglobulin IgA and IgG are main antibodies for mediating humoral immunity and important indexes for measuring the immune function of the body, and the content of the immunoglobulin can reflect the immune state of animals.

Research results show that compared with a basic diet treatment group, when 21-day-old broiler chickens added with 45-60g/t 5-ALA in the daily diet are 21 days old, the contents of immunoglobulin M (IgM) and immunoglobulin G (IgG) in serum (P <0.01) can be obviously improved (Table 6). But had no significant effect on immunoglobulin a (iga) (P > 0.05). Compared with a control group, the daily ration added with 30-100g/t 5-ALA can obviously improve the IgA content (P is less than 0.01), the serum IgA has various antibody activities, and the content of IgA in the blood of the test broiler chicken is obviously improved, which indicates that the immune function of the organism is improved to a certain extent and the immunity of the organism is enhanced; adding 30-70 g/t5-ALA to obviously improve the IgM content (P < 0.01); the addition of 30-100g/t obviously improves the immunoglobulin IgG (P < 0.01). As shown in FIG. 1, broiler chickens in the test group to which 50g/t 5-ALA was added had a detail-improving effect at the 42-day-old intestinal bleeding spots, as compared with the control group.

TABLE 6 Effect of the addition of 5-ALA to the diet on broiler serum immunoglobulins

Another important index for evaluating the immunological competence of the body is cytokine, which is a kind of bioactive substance synthesized and secreted by immune cells (such as monocyte, macrophage, T cell, B cell, NK cell, etc.) and some non-immune cells (endothelial cell, epidermal cell, fibroblast, etc.), and can be used as intercellular signal transmission molecule to mediate and regulate chronic inflammatory reaction.

As can be seen from table 7, the level of interferon-gamma (IFN- γ) in serum of 21-day-old broiler chickens was significantly increased by adding 5-ALA to the diet compared to the control group (P ═ 0.01). In addition, the addition of 5-ALA has a tendency to reduce serum tumor necrosis factor alpha (TNF-alpha) in 21-day-old broiler chickens (P < 0.1). The addition of 5-ALA also tended to reduce serum tumor necrosis factor α (TNF- α) in broilers at 42 days of age (P <0.1), but the addition of 5-ALA to the diet increased the IL-4 content in serum to some extent (P ═ 0.102). The 5-ALA added into the daily ration can reduce the content of the proinflammatory cytokines, improve the level of the anti-inflammatory cytokines, activate the innate and acquired immune systems of the organism and promote the improvement of the immune level of the organism.

TABLE 7 Effect of the addition of 5-ALA to the daily ration on the serum cytokines of broiler chickens

Therefore, the feed conversion ratio can be obviously reduced by selecting the 5-ALA added into the feed of the 1-42-day-old broiler chicken according to the amount of 30-100 g/ton; the immune organ index has a growing trend and has a certain positive effect on the establishment of the immune function of the broiler body and the maintenance of the environmental stability of an immune system; the serum catalase activity of the broiler chicken is obviously improved, the catalase activity in liver tissues of 21 days old can be obviously improved, and the content of malondialdehyde serving as a lipid oxidation marker can be obviously reduced; has the effect of obviously improving the activity of superoxide dismutase of 42-day-old broilers and the trend of reducing the content of MDA in liver tissues.

Intensive research shows that compared with a basic daily ration treatment group, the 21-day-old broiler chicken added with 45-60g/t 5-ALA in daily ration can obviously improve the IgM and IgG content in serum, and when the 21-day-old broiler chicken is 42 days old, the IgA content can be obviously improved. Meanwhile, the level of gamma-interferon (IFN-gamma) in the serum of 21-day-old broiler chicken can be obviously improved, the trend of serum tumor necrosis factor alpha (TNF-alpha) is reduced, and the IL-4 content in the serum is improved to a certain extent. The 5-ALA added into the daily ration can reduce the content of the proinflammatory cytokines, improve the level of the anti-inflammatory cytokines, activate the innate and acquired immune systems of the organism and promote the improvement of the immune level of the organism. In conclusion, 5-ALA added into the feed of 1-42-day-old broiler chickens in an amount of 45-60 g/ton is the optimal choice.

Example 25 feeding experiment of ALA on meat ducks and its influence on Productivity, serum Biochemical indicators and immune function

2.1 test materials

144 healthy 1-day-old Beijing duck male ducks with the weight of 50 +/-2.58 g close to each other are selected and randomly divided into 5 groups, each group has 6 repetitions, and each repetition has 8 ducks. The group design is as follows: t1 (basal diet), T2 (basal diet +30 g/T5-ALA), T3 (basal diet +40 g/T5-ALA), T4 (basal diet +70 g/T5-ALA), T5 (basal diet +100 g/T5-ALA). The test is carried out by adopting corn-soybean meal-miscellaneous meal type basic feed according to meat duck feeding standard (NY/T2122-2012).

TABLE 8 basic diet composition and nutritional ingredients for meat duck experiment

The vitamin premix is provided for each kilogram of feed: VA 10000IU, VD 32000 IU, VE 10IU, VK 32.5mg, VB11 mg, VB 26mg, VB 310 mg, VB 540 mg, VB 63 mg, VB110.3mg, VB120.01mg and biotin 0.12 mg.

The microelement premix is provided for each kilogram of feed: cu 8mg, Fe 80mg, Mn 60mg, Zn 40mg, Se 0.15mg, I0.353 mg.

2.2 measurement of indices and methods

2.2.1 growth Performance

When the meat ducks are 28 days old after the test, after the meat ducks are empty in stomach for 12 hours, weighing and setting the meat ducks repeatedly, and calculating the average weight, the average daily gain, the average daily feed intake, the feed weight ratio and the death and culling rate of the meat ducks aged 1-28 days.

2.2.2 sample Collection

When the meat ducks are 28 days old, 3 meat ducks are randomly selected from each repetition, jugular veins are collected, and serum biochemical indexes are measured. Collecting 10ml whole blood, placing in disposable vacuum blood collection tube (containing separation gel and coagulant), standing at room temperature for 1h, centrifuging at 3500r/min for 10min to prepare serum, and subpackaging in EP tube at-20 deg.C refrigerator for storage. The meat duck is slaughtered and dissected quickly after blood collection, a meat duck jejunum section is taken for 5cm, minced meat in the section is washed by normal saline, placed in an EP tube, placed in liquid nitrogen for quick freezing, and transferred to a refrigerator at minus 80 ℃ for standby.

2.2.3 serum antioxidant and immune index

Total antioxidant capacity (T-AOC), Catalase (CAT) activity, superoxide dismutase (T-SOD) activity and Malondialdehyde (MDA) content in serum are all determined by adopting Nanjing institute of bioengineering kit. Serum IgA, IgG and IgM contents were determined using an enzyme-linked immunosorbent assay (ELISA) kit, also purchased from Nanjing institute of bioengineering.

Jejunal TLR4 and cytokine content

Taking out jejunum segments frozen at-80 ℃, using a homogenizer to prepare tissue homogenate, and measuring the contents of TLR4, cytokines, namely tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta) and interleukin-6 (IL-6) by adopting corresponding ELISA kits which are purchased from Nanjing to build a bioengineering institute.

2.2.4 data processing

One-way ANOVA (one-way ANOVA) was performed on the test data using SPSS 19.0 software, and multiple comparisons were performed using the Duncan's method for indices of significant inter-group differences. The difference was considered significant with P <0.05 and very significant with P < 0.01.

2.3 results

2.3.15-influence of ALA on growth performance of meat duck

As can be seen from Table 9, compared with the group without 5-ALA, the weight, the daily average weight gain and the daily average feed intake (P is less than 0.05) of the minced meat duck of 28 days old can be obviously improved by adding 5-ALA, and the weight, the daily average weight gain and the daily average feed intake of the minced meat duck tend to increase with the increase of the additive amount, but the additive amounts are not obviously different; the weight ratio of the meat ducks aged 1-28 days can be obviously reduced by adding the 5-ALA, and the weight ratio of the meat ducks aged 40-100g/t of the 5-ALA is obviously different from that of the meat ducks not added with the 5-ALA (P is less than 0.05).

TABLE 95-influence of ALA on growth Performance of meat ducks (1-28d)

2.3.25-influence of ALA on antioxidant index of meat duck serum

As is clear from Table 10, the addition of 5-ALA at 30-100g/T increased the T-AOC, CAT, and T-SOD content in serum and decreased the MDA content, compared with the group without 5-ALA, and when the addition was 70-100g/T, the T-SOD content in serum was significantly increased (P < 0.05); when the addition amount is 40-100g/t, the content of MDA in serum can be obviously reduced compared with the group without 5-ALA (P is less than 0.05). The 5-ALA can improve the oxidation resistance of the meat duck body and reduce the MDA content in the body.

TABLE 105 influence of ALA on the antioxidant index of meat duck serum (1-28d)

2.3.35-influence of ALA on serum immunoglobulin content of meat duck

As is clear from Table 11, the addition of 5-ALA at 30-100g/t increased the content of IgA, IgG and IgM in the serum as compared with the group without 5-ALA, and the addition of 70-100g/t significantly increased the content of IgA in the serum as compared with the group without 5-ALA (P < 0.05); when the addition amount is 40-100g/t, the IgM content in serum can be obviously increased compared with the group without 5-ALA (P is less than 0.05). The 5-ALA can improve the immunoglobulin content of the meat duck body and increase the immune function of the body.

TABLE 115-Effect of ALA on serum immunoglobulin content of meat ducks

2.3.45-influence of ALA on content of TLR4 and cytokine in jejunum of meat duck

The inflammatory reaction is used as an important immune response of an organism, the animal health is damaged by excessive inflammatory reaction, and when livestock and poultry are infected by intestinal pathogenic bacteria, the TLR4 proinflammatory signal channel in the livestock and poultry is activated to generate inflammatory reaction, so that the expression of proinflammatory cytokines TNF-alpha, IL-1 beta, IL-6 and the like is increased, the normal function of an immune system is influenced, and the health of the livestock and poultry is threatened. As shown in Table 12, compared with the group without 5-ALA, the addition of 30-100g/t of 5-ALA can obviously reduce the secretory expression of the jejunal TLR4 (P is less than 0.05), and simultaneously reduce the transient secretion of proinflammatory factors TNF-alpha and IL-6 in the intestinal tract. The 5-ALA can improve the inflammatory reaction of the meat duck body and improve the body health.

TABLE 12 influence of the addition of 5-ALA to the daily ration on serum cytokines of meat ducks

Therefore, the 5-ALA added into the feed for the meat ducks aged 1-28 days according to the amount of 40-100 g/ton can obviously improve the daily gain, reduce the material weight ratio, increase the contents of T-AOC, CAT and T-SOD in serum and reduce the content of MDA, and when the adding amount is 70-100g/T, the content of T-SOD in serum can be obviously increased compared with a group without the 5-ALA; the content of IgA, IgG and IgM in the serum can be increased by adding 30-100g/t of 5-ALA, and the content of IgA in the serum can be remarkably increased compared with the group without adding 5-ALA when the adding amount is 70-100 g/t; the addition of 30-100g/t of 5-ALA can obviously reduce the secretory expression of the TLR4 in the jejunum and simultaneously reduce the transient secretion of proinflammatory factors TNF-alpha and IL-6 in the intestinal tract. In conclusion, the 5-ALA added into the feed for the meat ducks of 1-28 days old in an amount of 70-100 g/ton is the optimal choice.

Example 35 ALA feeding experiments on egg-laying hens and its effects on Productivity, Biochemical indicators of serum and immune function

3.1 test materials

500 healthy helan-brown laying hens aged 28 weeks were selected and randomly divided into 5 groups of 100 chickens per group, 4 replicates each of 25, and a test period of 49 days. The group design is as follows: t1 (basal diet), T2 (basal diet +30 g/T5-ALA), T3 (basal diet +40 g/T5-ALA), T4 (basal diet +70 g/T5-ALA), T5 (basal diet +100 g/T5-ALA). The test uses a corn-soybean meal-miscellaneous meal type basal diet, the composition and nutrient content of which are as follows:

TABLE 13 basic diet composition and nutritional ingredients

Per kg premix provided: VA 5200IU, VD 600IU, VE 10IU, VK 0.8mg, thiamine 0.8mg, riboflavin 2.3mg, calcium pantothenate 4.4mg, nicotinic acid 11mg, biotin 0.1mg, choline chloride 1100mg, folic acid 0.26mg, VB120.04mg, Cu 8mg, Fe 60mg, Mn 31mg, Zn 55mg, Se 0.1mg, I0.3 mg and antioxidant 100 mg.

3.2 measurement index and method

3.2.1 Performance

During the test period, the egg yield, egg weight, feed intake and egg breaking number are repeatedly recorded every day, and the egg yield, average egg weight, egg breaking rate, average daily feed intake and feed-egg ratio are counted by taking groups as units.

3.2.2 sample Collection

At the age of 49 days in the test, 3 laying hens are randomly selected from each repetition, and the blood is collected from the inferior pterygoid vein to determine the serum biochemical index. Collecting 10ml whole blood, placing in disposable vacuum blood collection tube (containing separation gel and coagulant), standing at room temperature for 1h, centrifuging at 3500r/min for 10min to prepare serum, and subpackaging in EP tube at-20 deg.C refrigerator for storage.

3.2.3 serum antioxidant and immune indices

Total antioxidant capacity (T-AOC), Catalase (CAT) activity, superoxide dismutase (T-SOD) activity and Malondialdehyde (MDA) content in serum are all determined by adopting Nanjing institute of bioengineering kit. Serum IgA, IgG and IgM contents were determined using an enzyme-linked immunosorbent assay (ELISA) kit, also purchased from Nanjing institute of bioengineering.

3.2.4 data processing

One-way ANOVA (one-way ANOVA) was performed on the test data using SPSS 19.0 software, and multiple comparisons were performed using the Duncan's method for indices of significant inter-group differences. The difference was considered significant with P <0.05 and very significant with P < 0.01.

3.3 results

3.3.15-ALA influence on layer production Performance

As can be seen from Table 14, the addition of 5-ALA tended to increase the feed intake and egg weight of the hens, but had no statistical difference (P >0.05) and decreased the feed-egg ratio (P >0.05) as compared with the group without 5-ALA; compared with a group without 5-ALA, the addition of 5-ALA can obviously reduce the egg breaking rate of the laying hens (P is less than 0.05); the addition of 5-ALA can improve the laying rate of the laying hens, and when the addition amount is 70-100g/t, the laying rate of the laying hens can be obviously improved (P is less than 0.05) compared with a group without the addition of 5-ALA.

TABLE 145-influence of ALA on layer production Performance (28w-35w)

3.3.25-ALA influence on egg-laying hen serum antioxidant index

As is clear from Table 15, the addition of 5-ALA at 30-100g/T increased the T-AOC, CAT, and T-SOD content in serum and decreased the MDA content compared to the group without 5-ALA, while the addition of 5-ALA at 30-100g/T significantly increased the T-AOC content in serum (P < 0.05); when the addition amount is 40-100g/T, compared with the group without 5-ALA, the content of T-SOD in serum can be obviously increased, and the content of MDA in serum can be reduced (P is less than 0.05). The 5-ALA can improve the oxidation resistance of the body of the laying hen and reduce the content of MDA in the body.

Influence of Table 155-ALA on anti-oxidation index of layer chicken serum

3.3.35-ALA influence on serum immunoglobulin content of egg-laying hen

As is clear from Table 16, the amounts of IgA, IgG and IgM in the serum were increased by adding 5-ALA at 30-100g/t as compared with the group without 5-ALA, and the amount of IgA in the serum was significantly increased (P <0.05) when the amount was 40-100g/t as compared with the group without 5-ALA; when the addition amount is 30-100g/t, the content of IgG and IgM in serum can be obviously increased (P is less than 0.05) compared with the group without 5-ALA. The 5-ALA can improve the immunoglobulin content of the laying hen body and increase the body immune function.

TABLE 165-influence of ALA on serum immunoglobulin content of layer hens

Therefore, 5-ALA added into the feed of the laying hens aged for more than 28 weeks in an amount of 30-100 g/ton tends to increase the feed intake and the egg weight of the laying hens, the feed-egg ratio tends to be reduced, the egg breaking rate of the laying hens can be obviously reduced, and the egg laying rate of the laying hens can be obviously improved when the adding amount is 70-100g/t compared with a group without the 5-ALA; the addition of 30-100g/T of 5-ALA can increase the content of T-AOC, CAT and T-SOD in serum, reduce the content of MDA and obviously increase the content of T-AOC in serum; the addition of 5-ALA at 30-100g/t can increase the content of IgA, IgG and IgM in the serum, and when the addition amount is 40-100g/t, the content of IgA in the serum can be obviously increased compared with the group without 5-ALA. In conclusion, 5-ALA added to feed for laying hens aged 28 weeks or more in an amount of 70-100 g/ton is the most preferable.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims (6)

1. A functional amino acid for improving the oxidation resistance and immunity of poultry, which is characterized in that the functional amino acid is 5-aminolevulinic acid, and the active ingredient of the 5-aminolevulinic acid is one or more of 5-aminolevulinic acid, 5-aminolevulinic acid derivatives or 5-aminolevulinic acid salts.

2. The application of the function of improving the oxidation resistance and the immunity of poultry based on 5-aminolevulinic acid in the poultry breeding industry is characterized in that one or more of 5-aminolevulinic acid, 5-aminolevulinic acid derivatives or 5-aminolevulinic acid salts are added into poultry feed, drinking water and medicines, and the daily intake of the poultry is 0.0001-1g calculated by 5-aminolevulinic acid.

3. The use according to claim 2, characterized in that one or more of 5-aminolevulinic acid, 5-aminolevulinic acid derivatives or 5-aminolevulinic acid salts are added into the complete feed for the poultry, and the addition amount of the complete feed for 1-42-day-old broiler chickens is 30-100 g/ton based on 5-aminolevulinic acid; the addition amount of the complete feed for meat ducks of 1-28 days old is 40-100 g/ton; the addition amount of the complete feed for laying hens aged over 28 weeks is 30-100 g/ton.

4. The application of the 5-aminolevulinic acid as the additive in the complete feed for 1-42-day-old broiler chickens as claimed in claim 3, wherein the additive amount of the 5-aminolevulinic acid is 45-60 g/ton.

5. The application of the feed as claimed in claim 3, wherein the addition amount of the 5-aminolevulinic acid in the complete feed for meat ducks aged 1 to 28 days is 70 to 100 g/ton.

6. The use according to claim 3, wherein the amount of 5-aminolevulinic acid added to the complete feed for laying hens aged 28 weeks or more is 70 to 100 g/ton.

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