CN112716942B - Indole and application of duck feed containing indole in maintaining duck intestinal health and/or preventing necrotic enteritis of ducks - Google Patents
Indole and application of duck feed containing indole in maintaining duck intestinal health and/or preventing necrotic enteritis of ducks Download PDFInfo
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Abstract
The present invention relates to the use of indoles and duck feed comprising the same for maintaining duck intestinal health and/or preventing duck necrotic enteritis, in particular caused by clostridium perfringens type a. The method has important significance for preventing epidemic diseases and ensuring the development of duck breeding industry by exploring, searching and applying the active substances to protect duck intestinal health, and provides a beneficial reference for solving bacterial drug resistance and antibiotic residue caused by abuse of antibiotics in the breeding industry.
Description
Technical Field
The invention relates to the technical field of green breeding of duck industry, in particular to the field of duck intestinal health, and more particularly relates to application of indole as a duck feed additive to maintaining duck intestinal health and/or preventing duck necrotic enteritis and a using method thereof.
Background
Necrotic enteritis of ducks is a digestive tract infectious disease and occurs all the year round, the typical clinical symptoms of the disease are that diseased ducks discharge dark brown tar-like feces accompanied with blood or intestinal mucosa tissues, the intestinal tissues at the later stage of the disease show mucosal ulcer and necrosis, the death rate of the disease can reach 30 percent at most, and huge loss is caused to the duck breeding industry in China [1] . The infection route of necrotic enteritis of ducks is mainly excrement discharged by sick ducks, so that feeding management, cleanness and sanitation are closely related to the occurrence of diseases.
Current research has found that one of the major therapeutic factors for necrotic enteritis in ducks is Clostridium perfringens (Clostridium perfringens), especially Clostridium perfringens type a. And the main lesion site of clostridium perfringens causing necrotic enteritis in ducks is the intestinal tract. It is well known that the intestinal tract is an important site for digestion and absorption of nutrients, and the intestinal mucosa is an important defense barrier against pathogenic microorganisms and virulence factors, and can maintain intestinal homeostasis [2] . The intact mucosa barrier in duck body is the basic condition for determining duck body health, and the timely repair and reconstruction of injured intestinal mucosa plays a key role in preventing pathogen from continuously invading [3] . Under pathogenic stress, clostridium perfringens can destroy the compact structure among animal intestinal epithelial cells, influence the barrier function of intestinal tracts and trigger systemic inflammatory response. For example, enterotoxin produced by the bacterium perfringens is capable of specifically binding to Claudin-3 to cause structural disruption of Claudin and induce apoptosis [4] . Endotoxin Escherichia coli Lipopolysaccharide (LPS) causes redistribution and combination of intracellular tight junction proteins Occludin, ZO-1 and Claudins [5] Further inducing the production of inflammatory mediators such as TNF-alpha and the like to induce the apoptosis of intestinal mucosa cells and damage the connection among the intestinal cells and the barrier integrity of the intestinal mucosa [6] 。
In the prior art, necrotic enteritis of ducks is mainly prevented and treated by medicines, and meanwhile, feeding management of duck groups is strengthened. For example, sick ducks may be treated with trimethoprim in combination with 0.2% florfenicol in a drinking water [7] (ii) a The antibiotics such as streptomycin, kanamycin and the like can be injected into diseased ducks to treat necrotic enteritis of the ducksEffective prevention and cure [8] (ii) a The compound preparation of coptis chinensis, phellodendron amurense and dark plum powder has better effect on treating duck necrotizing enteronitis [9] (ii) a The incidence rate of necrotic enteritis of ducks can be effectively reduced by reasonable supplementary feeding of duck groups and the measures of strict disinfection and isolation. However, with the widespread use and abuse of veterinary antibiotics, pathogen resistance has increased, which has led to a gradual decrease in the efficacy of the antibiotics.
In the face of increasingly serious germ drug resistance, the concept of 'no resistance' or 'resistance reduction' food is increasingly deep, so that an active substance is urgently needed to be found to protect duck intestinal health and reduce antibiotic use.
Disclosure of Invention
As described above, there is a need in the art for an active substance that can effectively protect duck intestinal health, and can prevent duck necrotic enteritis without the use of antibiotics. Thus, the present invention provides a solution to the problems associated with duck intestinal health and/or duck necrotic enteritis.
The inventors of the present application have found that when an effective amount of a food grade indole is used as an additive to a duck feed, it is effective to maintain duck intestinal health with low or no antibiotics during duck feeding and to prevent necrotic enteritis in the duck caused by clostridium perfringens, especially clostridium perfringens type a. Based on this finding, the present inventors have completed the present invention.
Accordingly, in a first aspect of the invention, the invention provides the use of an indole for the manufacture of a medicament for maintaining duck intestinal health and/or preventing duck necrotic enteritis.
In a second aspect, the present invention provides an indole premix for duck feed, comprising indole and a carrier, wherein the indole is 0.5% to 0.8% by weight of the indole premix.
In a third aspect, the present invention provides a duck feed comprising 5% by weight of the indole premix of the second aspect based on the weight of the duck feed.
The invention has the advantages that: the invention discovers for the first time that the addition of an effective amount of indole into duck feed can maintain the health of duck intestinal tracts, and prevent necrotic enteritis of ducks caused by clostridium perfringens, especially clostridium perfringens type A under the condition of 'no resistance' or 'low resistance', so that the use of antibiotics during feeding of ducks can be effectively reduced, the antibiotic residue and bacterial drug resistance caused by the antibiotic residue are avoided, and further the economic benefit of duck farms is improved and the ecological environment is protected.
Detailed Description
The present invention will be described in detail below. It is to be understood that the following description is intended to illustrate the present invention by way of example only and is not intended to limit the scope of the invention, which is defined by the appended claims. Also, it is understood by those skilled in the art that modifications may be made to the technical aspects of the present invention without departing from the spirit and gist of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.
As mentioned above, there is a need in the art for an active substance that can maintain duck intestinal health in the absence of "resistance" or "low resistance" and thereby prevent necrotic enteritis in ducks caused by clostridium perfringens, especially clostridium perfringens type a, during duck feeding.
In a first aspect of the invention, there is provided the use of indole in the manufacture of a medicament for maintaining duck intestinal health and/or preventing duck necrotic enteritis.
The term "gut health" encompasses a number of physiological and functional characteristics including efficient digestive absorption of nutrients, stable microflora, effective immune status, intact barrier function and sensitive neuroendocrine function, among others, being an ability to maintain normal physiological and homeostasis of the gut, resistance to infections and non-infectious diseases [10] . The unbalanced intestinal health of the waterfowl can influence the digestion, absorption and utilization of nutrients by the waterfowl, further influence the growth, development and production of the waterfowl and cause unnecessary economic loss in the production process of the waterfowl. In the production practice, the intestinal health can be regulated and maintained by regulating and controlling the growth and repair of intestinal mucosa, maintaining the microbial community balance, regulating the intestinal immune state and the like, and the intestinal health imbalance is avoided.
The term "necrotic enteritis in ducks", also known as "rotten bowel disease", is an acute infectious disease caused by clostridium perfringens, especially by clostridium perfringens type a, occurring all the year round, with autumn and winter being the high-incidence season. It is clinically characterized by depression, loss of appetite, inability to stand, diarrhea, and often sudden death. Once the duck group is infected with the disease, the morbidity and mortality of the duck group are high, so that the duck group becomes one of the main diseases causing economic loss of the duck breeding industry. In the current production practice, necrotic enteritis in ducks is mainly prevented and treated by medicines, such as trimethoprim and 0.2% florfenicol combined drug, streptomycin, kanamycin and other antibiotics are injected, and coptis chinensis, phellodendron amurense and dark plum powder compound preparations and the like are applied. However, these treatments generally involve the use of veterinary antibiotics, and the widespread use and abuse of veterinary antibiotics has resulted in increased resistance to clostridium perfringens, resulting in a gradual decline in antibiotic efficacy.
In a particular embodiment, the necrotic enteritis in ducks is caused by Clostridium perfringens (Clostridium perfringens).
Clostridium perfringens is a gram-positive rod-shaped anaerobic bacterium known for its ability to break down sugars in muscle and connective tissues to produce large amounts of gas and to form a capsule in the body. Clostridium perfringens is widely distributed in the natural world, in the intestines of healthy animals and humans, and belongs to a opportunistic pathogen. In healthy animals, clostridium perfringens has low pathogenicity, but can cause diseases such as necrotic enteritis of livestock and poultry when factors such as climate, nutrition, immunity, intestinal flora imbalance and feeding management change.
In a preferred embodiment, the necrotic enteritis in ducks is caused by clostridium perfringens type a. Clostridium perfringens type a is one of the main pathogenic bacteria causing necrotic enteritis in various animals, and its causative agent is alpha toxin produced by the bacteria.
In a specific embodiment, the daily dose of indole is between 0.0225g and 0.04 g.
Indole is a metabolite of tryptophan, which is widely found in meat, fish and eggs and is an essential amino acid for animals. In vivo symbiotic bacteria (such as Escherichia coli and Vibrio) decompose tryptophan into indole under the action of tryptophanase. The inventor unexpectedly finds that the tryptophan metabolite indole can influence the barrier homeostasis of small intestine epithelial cells of ducks and plays a role in the repair and reconstruction of the intestinal epithelial barrier.
In a second aspect, the present invention provides an indole premix for duck feed comprising indole and a carrier, wherein the indole is 0.5% to 0.8% by weight of the indole premix.
The carrier can be any substance which can be used as a premix carrier in the feed field, such as bran, zeolite powder, rice hull powder and bran powder. In one embodiment, the carrier may be bran. Bran is a by-product of wheat processing flour and is therefore also known as "oatmeal", which is a mally yellow color, typically in the form of flakes or flour. Bran is used in various breeding feeds due to its rich content of cellulose, vitamins and minerals.
In a preferred embodiment, wherein the indole is 0.5% by weight of the indole premix.
In yet another preferred embodiment, wherein the indole has a particle size of no more than 0.425 mm.
In a third aspect, the invention provides a duck feed comprising 5% by weight of the indole premix of the second aspect based on the weight of the duck feed.
In a specific embodiment, the duck feed further comprises 95% by weight of duck feed of duck powder, the duck powder comprising the following ingredients by weight of duck powder:
45 to 55 percent of corn by weight percent,
3 to 8 percent of bean cakes,
5 to 15 percent of oat, and the oat is rice,
15 to 20 percent of wheat, and the balance of wheat,
8 to 15 percent of bran coat,
3 to 6 percent of fish meal,
0.1 to 3 percent of bone meal,
from 0.1% to 3% of shell powder, and
0.1% to 1% of common salt.
In a particular embodiment, the duck powder comprises the following ingredients by weight of duck powder: 50% of corn, 5% of bean cake, 10% of oat, 17% of wheat, 11% of bran, 4.5% of fish meal, 1% of bone meal, 1% of shell powder and 0.5% of salt.
In a specific embodiment, the duck powder is additionally added with 0.005% to 0.015% of lactic acid bacteria by weight of the duck powder.
In a preferred embodiment, the duck powder is additionally added with 0.01% of lactic acid bacteria by weight of the duck powder.
In a more specific embodiment, the lactic acid bacterium is Lactobacillus plantarum (Lactobacillus plantarum).
The lactic acid bacteria of the present invention may be a commercial product. The lactobacillus added into the duck feed can regulate the gastrointestinal flora of a duck body, keep the microecological balance of the gastrointestinal tract, inhibit the reproduction of intestinal putrefying bacteria, further improve the digestibility of food, enhance the immunity of the body and improve the health level of the body. Without wishing to be bound by theory, when lactic acid bacteria are used in combination with the indoles of the present invention, they are synergistically able to regulate the gastrointestinal flora of the duck organism, maintain the microecological balance of the gastrointestinal tract, inhibit the proliferation of intestinal putrefying bacteria, thereby maintaining duck intestinal health and/or preventing duck necrotic enteritis. This is because on the one hand bacteriocins produced by lactic acid bacteria, especially lactobacillus plantarum, can be correlated with strain Quorum Sensing (QS), which in turn affects pathogen virulence factor expression; on the other hand, lactic acid bacteria are capable of decomposing tryptophan itself into indole. The exogenously added indole and the indole decomposed by the lactobacillus can synergistically inhibit the propagation of pathogenic bacteria.
In yet another specific embodiment, the ground particle size of the duck powder is 3mm to 5 mm.
In yet another preferred embodiment, the ground particle size of the duck powder is 4 mm.
The invention has the advantages that: the inventor finds that the effective amount of indole added into the duck feed can maintain the duck intestinal health, and prevent necrotic enteritis of the duck under the condition of 'no resistance' or 'low resistance', so that the use of antibiotics during feeding of the duck can be effectively reduced, the antibiotic residue and bacterial drug resistance caused by the antibiotic residue are avoided, the economic benefit of a duck farm is improved, and the ecological environment is protected. Therefore, the invention has important significance for exploring, searching and applying active substances to protect duck intestinal health, preventing epidemic diseases and ensuring the development of duck breeding industry, and provides beneficial reference for solving bacterial drug resistance and antibiotic residue caused by antibiotic abuse in the breeding industry.
Examples
The following examples will demonstrate the effect of indole as a duck feed additive in maintaining duck intestinal health and/or preventing duck necrotizing enteritis caused by clostridium perfringens, with a gaoyou duck as the feeding subject.
The test methods employed in the present invention are all conventional methods unless otherwise specified, and the test materials used in the following examples are all purchased from conventional stores unless otherwise specified. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It should be noted that the terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The foregoing summary, as well as the following detailed description, is intended merely to be illustrative of the invention and is not intended to be in any way limiting. The scope of the invention is to be determined by the appended claims without departing from the spirit and scope of the invention.
Example 1: preparation of duck feed
Preparing duck feed by the following steps:
s1: crushing food-grade indole with an ultramicro machine, and sieving with a 40-mesh woven sieve to obtain indole with particle size below 0.425 mm;
s2: respectively taking the indole obtained by the step S1 based on the weight of the indole premix given in the following table 1, and mixing the indole with bran in a corresponding weight ratio to obtain indole premixes with different indole contents;
s3: crushing and uniformly mixing the components in the weight of the duck powder given in the following table 1 to obtain self-prepared duck powder; optionally, according to the data given in table 1 below, lactobacillus plantarum, additionally added by weight of duck powder;
s4: taking 50g of each indole premix obtained in the step S2 and 950g of each duck powder prepared in the step S3, and adding for multiple times and mixing step by step to obtain a control group 1 and a control group 2 (containing no indole), a toxicity test group (the weight percentage of the indole is 1 percent of the weight of the indole premix), and a test group 1-6 feed (the weight percentage of the indole is the weight of the indole premix, and the details are shown in the following table 1) given in the following table 1.
Example 2: toxicity test of indole on Duck
Toxicity tests were performed on 20 22-day-old Gaoyou ducks (10 each male and female) to see if indole is toxic to ducks. Specifically, gaoyou ducks (5 male and female ducks in each group) were fed with the feed (containing no indole) of the control group 1 and the feed (containing 0.05% by weight of indole) of the toxicity test group prepared in example 1 and were continuously fed for 7 days. After 7 days, three male ducks and female ducks of the control group and the toxicity test group are dissected randomly.
The internal organs obtained in the toxicity test group were not changed from the control group by visual observation. The biochemical indexes of the blood of the control group and the toxicity test group are also detected, and the results are shown in the following table 2. The results show that the data of the control group and the toxicity test group are not significantly different. Therefore, the indole added in the feed is safe for the ducks under the condition that the weight percentage is not more than 0.05%.
Table 2: blood biochemical index data of ducks of control group and toxicity test group
Note: in the table, the same letter indicates that there is no statistical difference p > 0.05 between the same row of data, and different letters indicate that there is a statistical difference p < 0.05 between the same row of data.
Example 3: indole intestinal health promotion test
The test adopts single-factor design, and the variable is only the content of indole in the duck feed.
Specifically, 280 Gaoyou ducks with similar weight and age of 18 days are randomly selected and divided into 4 groups, and the male ducks and the female ducks are half of the male ducks and the female ducks respectively.
The pre-feeding period was 4 days and the feed was control 1 feed of table 1 in example 1, i.e. no indole; the test period was 28 days, starting at 22 days of age and ending at 49 days of age. During the whole test period, all ducks freely take powder and drinking water, and the daily ration and feeding management refer to local standards of Gaoyou duck feeding technical regulations (DB 32/T910-2006). The daily feed intake of ducks is usually 90g to 100 g.
The test was divided into control group 1 and test group 1, test group 2 and test group 3, and the respective ingredients of the feed were as shown in the respective groups in table 1 above. After the test is finished, male and female ducks are randomly selected from the control group and the test group, slaughtered respectively, and cecal tissues are collected by using an RT-PCR method and a TaqMan probe method which are well known in the art [11] Determination of the number of Clostridium perfringens in the intestine, expressed as the common logarithm of the number of Clostridium perfringens per gram of content [ lg (number of copies/g)]And counting the incidence rate of necrotic enteritis of each group of ducks. The results of the control group 1 and the test groups 1 to 3 are shown in Table 3 below (n in the table is the number of ducks in each group).
Table 3: number and incidence of Clostridium perfringens in each group
Note: in the table, the same letter indicates that there is no statistical difference p > 0.05 between the same column data, and different letters indicate that there is a statistical difference p < 0.05 between the same column data.
The test results in table 3 show that there was no statistically significant difference in the numbers of clostridium perfringens type a in the control and test groups (i.e., test group 1) when fed a lower dose of indole (0.2% by weight of indole premix), and the incidence of the test group was reduced compared to the control group, but there was no significant difference (p > 0.05); when the control group and the test group (namely the test group 2 and the test group 3) are fed with indole at higher doses (0.5 percent and 0.8 percent by weight of the indole premix), the number of clostridium perfringens type A in the control group and the test group is statistically different (p is less than 0.05), and the incidence rate of necrotic enteritis of ducks is also remarkably lower than that in the control group. Of the three test groups, clostridium perfringens of test group 2 having an indole content of 0.5% by weight based on the indole premix was the lowest in number, and duck necrotic enteritis of test group 3 having an indole content of 0.8% by weight based on the indole premix was the lowest in incidence, which indicates that the effect of 0.5% to 0.8% by weight of the indole premix in the feed for ducks was the best for maintaining the health of duck intestinal tracts.
Example 4: synergistic gut health test with indole and lactobacillus plantarum
The test adopts a two-factor design, and the variable is that the duck feed contains or does not contain indole and lactobacillus plantarum.
Similar to example 3, 280 senior post ducks of 18 days old with similar body weight were randomly selected and divided into 4 groups:
control group 1: contains lactobacillus plantarum (0.01%) only and no indole;
control group 2: contains neither lactobacillus plantarum nor indole;
test group 2: containing both lactobacillus plantarum (0.01%) and indole (0.5%);
test group 4: only indole (0.5%) was contained and no lactobacillus plantarum was contained.
The components of the duck powder of the control group and the test group are completely the same, and the differences are only indole and lactobacillus plantarum, and the contents of indole and lactobacillus plantarum are shown in table 1 above.
The pre-feeding period was 4 days, the feed was the feed used in control 1 of example 1, i.e. comprising lactobacillus plantarum only and no indole, and the experimental period was 28 days, starting at 22 days of age and ending at 49 days of age. During the whole test period, all ducks freely feed powder and drinking water, and the daily ration and feeding management refer to local standards of Gaoyou duck feeding technical regulations (DB 32/T910-2006). The daily feed intake of ducks is usually 90g to 100 g.
After the test is finished, male ducks and female ducks are randomly selected from a control group and a test group, slaughtered respectively, cecum tissues are adopted, the quantity of clostridium perfringens in intestinal tracts is determined by using an RT-PCR method and a TaqMan probe method, the quantity is expressed as a common logarithm of the quantity of clostridium perfringens in each gram of content [ lg (copy number/g) ], and the incidence rate of necrotic enteritis of the ducks of each group is counted. The results of the control groups 1 and 2 and the test groups 2 and 4 are shown in the following table 4 (in the table, n is the number of ducks in each group).
Table 4: number and incidence of Clostridium perfringens in groups
Note: in the table, the same letter indicates that there is no statistical difference p > 0.05 between the data in the same column, and different letters indicate that there is a statistical difference p < 0.05 between the data in the same column.
As shown in the test results in table 4, comparing the results of the control group and the test group, it can be seen that the addition of indole and lactobacillus plantarum can not only significantly reduce the number of clostridium perfringens, but also reduce the incidence of necrotic enteritis in ducks under the condition that other components are identical. And test group 2 containing both lactobacillus plantarum and indole, whether clostridium perfringens numbers or duck necrotic enteritis incidence was significantly better than control groups 1 and 2 and test group 4 containing only indole, which showed a synergistic effect of both lactobacillus plantarum and indole for maintaining intestinal health.
While embodiments of the present invention have been described, the present invention is not limited to the above-described embodiments, which are intended to be illustrative rather than limiting, and many modifications may be made by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.
Reference to the literature
[1] Analysis, diagnosis and prevention scheme of necrotic enteritis of populus, ducks and geese [ J ], modern animal science and technology, 2019, 02: 144-145;
[2]Blikslager AT,Moeser AJ,Gookin JL et al,Restoration of barrier function in injured intestinal mucosa[J],Physiological reviews,2007,87(2): 545-564;
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Claims (14)
1. Use of indole in preparation of medicine for preventing necrotic enteritis of duck.
2. The use of claim 1, wherein the necrotic enteritis is caused by clostridium perfringens (c)Clostridium perfringens) And (4) causing.
3. The use of claim 1 or 2, wherein the duck necrotizing enterocolitis is caused by clostridium perfringens type a.
4. Use according to claim 1 or 2, wherein the daily dose of indole is between 0.0225g and 0.04 g.
5. Use of indole for the preparation of a premix for duck feed for the prevention of necrotic enteritis, the premix comprising indole and a carrier, wherein the indole is present in an amount of 0.5% to 0.8% by weight of the premix.
6. Use according to claim 5, wherein the indole is 0.5% by weight of the premix.
7. Use according to claim 5 or 6, wherein the indole has a particle size of no more than 0.425 mm.
8. A duck feed for preventing necrotic enteritis in ducks, comprising 5% of the premix of any one of claims 5-7 by weight of the duck feed and 95% of duck powder by weight of the duck feed, the duck powder comprising the following ingredients by weight of the duck powder:
45 to 55 percent of corn by weight percent,
3 to 8 percent of bean cakes,
5 to 15 percent of oat, and the oat is,
15 to 20 percent of wheat, and the balance of wheat,
8 to 15 percent of bran coat,
3 to 6 percent of fish meal,
0.1 to 3%% of bone meal,
from 0.1% to 3% of shell powder, and
0.1% to 1% of common salt.
9. The duck feed of claim 8, wherein the duck powder comprises the following ingredients by weight of duck powder: 50% of corn, 5% of bean cake, 10% of oat, 17% of wheat, 11% of bran, 4.5% of fish meal, 1% of bone meal, 1% of shell powder and 0.5% of salt.
10. The duck feed according to claim 8 or 9, wherein 0.005% to 0.015% of lactic acid bacteria by weight of duck powder is added to the duck powder.
11. The duck feed according to claim 10, wherein 0.01% of lactic acid bacteria by weight of duck powder is added to the duck powder.
12. The duck feed according to claim 10, wherein the lactic acid bacteria are lactobacillus plantarum (lactobacillus)Lactobacillus plantarum)。
13. The duck feed as claimed in claim 8, wherein the ground duck powder has a particle size of 3mm to 5 mm.
14. The duck feed of claim 13, wherein the duck powder has a particle size of 4 mm.
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