CN113508870B - Feed containing fermented pepper straw - Google Patents
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/105—Aliphatic or alicyclic compounds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/174—Vitamins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/26—Compounds containing phosphorus
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/30—Oligoelements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
The invention relates to a feed containing fermented pepper straws, which consists of bacillus subtilis fermented pepper straw powder and basic ration, and the feed effect of the fermented pepper straw powder on the broiler is researched by measuring the production performance, the immunity index and the intestinal microbial diversity of the broiler so as to discuss the action mechanism of the fermented pepper straws on livestock and poultry and provide a reference for the reasonable application of the pepper straws in the animal husbandry.
Description
Technical Field
The invention belongs to the field of animal husbandry feeds, and particularly relates to a feed containing fermented pepper straws.
Background
At present, straws are mostly used for animal breeding in a fermentation mode, a good breeding effect is obtained, the production performance of ruminants is improved, the breeding cost is reduced, microbial fermentation is not adversely affected on the health of the ruminants, lignin in the straws can be effectively degraded, the nutritional value of feed is improved, and the feeding value of the straws is improved.
The gut flora is an important component of the digestive system of all animals, and changes in dietary composition and nutritional levels have a significant effect on the number and variety of gut microflora, thereby affecting the ability of the gut to digest and absorb nutrients. However, the use of straw in chicken feed is still a controversial issue due to the characteristics of the digestive system of the chicken. At present, few test reports about the fermentation of the pepper straw powder for raising the chickens exist.
Therefore, the bacillus subtilis fermented pepper straw powder is added into basic ration, the feeding effect of the fermented pepper straw powder on the broiler is researched by measuring the production performance, the immunity index and the intestinal microbial diversity of the broiler, the action mechanism of the fermented pepper straw on livestock is discussed, and reference is provided for reasonable application of the pepper straw in the animal husbandry.
Disclosure of Invention
The invention aims to provide a feed containing fermented pepper straws.
The feed consists of basic ration and fermented pepper straw powder, wherein the basic ration consists of the following components in parts by weight: 60-70 parts of corn, 20-30 parts of soybean meal, 3-5 parts of soybean oil, 0.5-1 part of stone powder, 0.1-0.5 part of salt, 0.1-1 part of calcium hydrophosphate and 2-6 parts of premix; the fermented pepper straw powder accounts for 1-4% of the basic ration in parts by weight.
In a preferred embodiment of the method of the invention,
the feed consists of basic daily ration and fermented pepper straw powder, wherein the basic daily ration consists of the following components in parts by weight: 62-68 parts of corn, 22-28 parts of soybean meal, 3.2-4.5 parts of soybean oil, 0.6-0.9 part of stone powder, 0.2-0.4 part of salt, 0.2-0.8 part of calcium hydrophosphate and 3-5 parts of premix; the fermented pepper straw powder accounts for 1.5-3% of the basic ration in parts by weight.
It is further preferred that the first and second liquid compositions are,
the feed consists of basic daily ration and fermented pepper straw powder, wherein the basic daily ration consists of the following components in parts by weight: 66.9 parts of corn, 24 parts of soybean meal, 3.5 parts of soybean oil, 0.8 part of stone powder, 0.3 part of salt, 0.5 part of calcium hydrophosphate and 4 parts of premix; the additive is fermented pepper straw powder, and the weight of the fermented pepper straw powder is 2% of that of the basic ration.
The preparation method of the fermented chili straw powder comprises the following steps:
1) Smashing pepper straws to 1-5cm in length;
2) Resuscitating and expanding the bacillus subtilis strain to prepare a bacterial liquid with the bacterial quantity more than or equal to 1 multiplied by 1010 cfu/mL;
3) Mixing water and straw at a ratio of 0.5-1.5, adding bacterial solution, stirring, sealing, and fermenting at 20-28 deg.C for 30-40 days to obtain the final product.
In a preferred embodiment of the method of the invention,
the preparation method of the fermented pepper straw powder comprises the following steps:
1) Smashing pepper straws to 1.5-3cm in length;
2) Reviving and enlarging culturing Bacillus subtilis strain to obtain the strain with bacterial quantity not less than 1 × 10 10 cfu/mL bacterial liquid;
3) Mixing water and straw at a ratio of 0.8-1.2, adding 10% bacteria solution, stirring, sealing, and fermenting at 22-26 deg.C for 32-38 days to obtain the final product.
It is further preferred that the first and second liquid crystal compositions,
the preparation method of the fermented pepper straw powder comprises the following steps:
1) Smashing pepper straws to 2cm in length;
2) Reviving and enlarging culturing Bacillus subtilis strain to obtain the strain with bacterial quantity not less than 1 × 10 10 cfu/mL bacterial liquid;
3) Uniformly stirring water and straws in a ratio of 1.
The pepper straw is applied to animal husbandry.
The fermented pepper straw powder is applied to broiler chicken feeding.
The fermented chili straw powder is applied to the aspects of broiler chicken production performance, immune function, serum antioxidant index and cecal microorganisms.
The fermented pepper straw powder has mellow fragrance or acid fragrance.
The "parts" in the present invention may be units known in the art such as kg and g.
The invention has the following advantages and beneficial effects:
1. according to the invention, the fermented pepper straws are added into the feed, so that the daily gain of the broiler chicken is obviously improved, and the bursal index, the thymus index and the spleen of the broiler chicken are respectively improved by 180.95%, 70.82% and 27.77%. The serum immunoglobulin IgG has larger influence, and is increased by 39.49 percent compared with a control group; the content of glutathione peroxidase is obviously improved by 32.27 percent compared with that of a control group, and the content of malondialdehyde is obviously reduced by 22.27 percent.
2. The invention classifies and identifies the broiler cecal phylum and the genus level sequence, and the dominant phylum of the cecal is bacteroidetes and firmicutes; the dominant bacteria are Bacteroides and Clostridium molesta. The relative abundance of the phylum of the treatment group, i.e., the phylum of the traceobacter and the phylum of the cyanobacteria, is obviously lower than that of the control group at the level of the phylum of the cecum. The relative abundance of Bacteroides is obviously reduced at the cecal level and is obviously lower than that of a control group, and the relative abundance of coprococcus is obviously improved.
3. According to the invention, through the test of the influence of the fermented pepper straw added into the feed on the production performance of the broiler, the result shows that the average daily gain can be obviously increased and the feed-weight ratio can be reduced by properly adding 2% of the fermented straw feed into the daily ration of the broiler in the growth period.
4. According to the experiment on the influence of the fermented pepper straws added into the feed on the immune function of the broiler chicken, the result shows that the thymus, spleen and bursal disease indexes can be obviously improved by adding the fermented pepper straws into the daily ration of the broiler chicken, wherein the bursal disease has the most obvious effect and is improved by 180.95% compared with a blank group, and the bacillus subtilis fermented pepper straws can obviously improve the immune level of the broiler chicken, so that the immune capacity of the broiler chicken on diseases is improved; the experiment also finds that the level of IgG in the broiler serum can be improved by adding the fermented straws into the basic feed, and is improved by 39.49% compared with a control group, and the result shows that the serum immunoglobulin IgG is greatly influenced by adding the pepper straws into the feed.
5. According to the invention, through a test of the influence of the fermented pepper straws added into the feed on broiler serum antioxidant indexes, the result shows that the contents of T-AOC, SOD, POD and CAT in broiler serum fed with the straw-added feed have an increasing trend compared with a control group, the content of GSH-PX is obviously increased by 32.27% (P < 0.05) compared with the control group, and the result shows that the addition of the fermented pepper straws into broiler daily ration can improve the activities of SOD and GSH-PX of broiler antioxidant enzymes, further improve the total antioxidant capacity (T-AOC) of an organism and reduce the activity of malonaldehyde.
6. According to the invention, through the test of the influence of the fermented pepper straws added into the feed on the broiler cecal microorganisms, the result shows that the relative abundance of the broiler cecal bacteroides is obviously reduced compared with that of a control group, and the relative abundance of the clostridium tenderizer is obviously higher than that of the control group, so that the bacillus subtilis is fed to ferment the pepper straws, so that the inflammation of an organism can be reduced, the intestinal flora structure of broilers is improved, the etiology of intestinal diseases is prevented, the intestinal health is favorably maintained, and the growth of the broilers is promoted. Studies have shown that bacteroidetes groups are beneficial for the normal development of the gastrointestinal tract and that they produce butyrate, which is the major product of colonic fermentation and has anti-tumor and gut health-preserving properties.
7. The feed additive consists of bacillus subtilis fermented pepper straw powder and basic ration, and is used for researching the feed effect of the fermented pepper straw powder on broilers by measuring the production performance, the immunity index and the intestinal microbial diversity of the broilers so as to discuss the action mechanism of the fermented pepper straw on livestock and poultry and provide reference for reasonable application of the pepper straw in the animal husbandry.
Drawings
FIG. 1 dilution curve for sequencing of 16S r RNA gene from caecum microorganism (A represents control group, B represents test group);
FIG. 2 is a sequencing scale abundance line of 16S r RNA gene of caecum microorganism (A represents a control group, and B represents a test group);
FIG. 3NMDS analysis (each point in the graph represents a sample, the distance between the point represents the degree of difference, and Stress is 0.2 less, indicating that NDNS can accurately reflect the degree of difference between samples).
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
Example 1 feed formulation
The fermented pepper straw feed formula comprises the following components: 66.9g of corn, 24g of bean pulp, 3.5g of soybean oil, 0.8g of stone powder, 0.3g of salt, 0.5g of calcium hydrophosphate and 4g of premix; 2g of fermented pepper straw powder.
Example 2 feed formulation
The formula of the fermented pepper straw feed comprises the following components: 60g of corn, 20g of soybean meal, 3g of soybean oil, 0.5g of stone powder, 0.1g of salt, 0.1g of calcium hydrophosphate and 2g of premix; 1g of fermented pepper straw powder.
Example 3 feed formulation
The formula of the fermented pepper straw feed comprises the following components: 70g of corn, 30g of soybean meal, 5g of soybean oil, 1g of stone powder, 0.5g of salt, 1g of calcium hydrophosphate and 6g of premix; 4g of fermented pepper straw powder.
Example 4 feed formulation
The formula of the fermented pepper straw feed comprises the following components: 63g of corn, 22g of soybean meal, 3.2g of soybean oil, 0.6g of stone powder, 0.2g of salt, 0.2g of calcium hydrophosphate and 3g of premix; 1.5g of fermented pepper straw powder.
Example 5 feed formulation
The fermented pepper straw feed formula comprises the following components: 68g of corn, 28g of soybean meal, 4.5g of soybean oil, 0.9g of stone powder, 0.4g of salt, 0.8g of calcium hydrophosphate and 5g of premix; 3g of fermented pepper straw powder.
Example 6 feed formulation
The formula of the fermented pepper straw feed comprises the following components: 65g of corn, 25g of soybean meal, 4.2g of soybean oil, 0.7g of stone powder, 0.4g of salt, 0.3g of calcium hydrophosphate and 4g of premix; 3.5g of fermented pepper straw powder.
Example 7 feed formulation
The formula of the fermented pepper straw feed comprises the following components: 69g of corn, 29g of soybean meal, 4.9g of soybean oil, 0.7g of stone powder, 0.3g of salt, 0.7g of calcium hydrophosphate and 5.5g of premix; 3g of fermented pepper straw powder.
The fermented pepper straw powder in the formulation of examples 1-7 was prepared according to the preparation method of the feed in examples 8-15, respectively.
Example 8 preparation of fermented Capsicum straw powder
1) Smashing pepper straws to 2cm in length;
2) Recovering Bacillus subtilis strain, and enlarging culturing to obtain strain amount =1 × 10 10 cfu/mL bacterial liquid;
3) Uniformly stirring water and straws in a ratio of 1.
Example 9 preparation of fermented Capsicum straw powder
1) Smashing pepper straws to 1cm in length;
2) Recovering Bacillus subtilis strain, and enlarging culturing to obtain strain amount =1 × 10 10 cfu/mL bacterial liquid;
3) Uniformly stirring water and straws in a ratio of 0.5.
Example 10 preparation of fermented Capsicum straw powder
1) Smashing pepper straws to 5cm in length;
2) Reviving and enlarging culturing Bacillus subtilis strain to obtain the strain with bacterial quantity not less than 1 × 10 10 cfu/mL bacterial liquid;
3) Uniformly stirring water and straws in a ratio of 1.5.
Example 11 preparation of fermented Hot Pepper straw powder
1) Smashing pepper straws into 3cm in length;
2) Reviving and enlarging culturing Bacillus subtilis strain to obtain the strain with bacterial quantity not less than 1 × 10 10 cfu/mL bacterial liquid;
3) Mixing water and straw at a ratio of 0.8.
Example 12 preparation of fermented Capsicum straw powder
1) Smashing pepper straws to 4cm in length;
2) Reviving and enlarging culturing Bacillus subtilis strain to obtain Bacillus subtilis with bacterial amount not less than 1 × 10 10 cfu/mL bacterial liquid;
3) Uniformly stirring water and straws in a ratio of 1.2.
Example 13 preparation of fermented Capsicum straw powder
1) Smashing pepper straws into 3cm in length;
2) Reviving and enlarging culturing Bacillus subtilis strain to obtain the strain with bacterial quantity not less than 1 × 10 10 cfu/mL bacterial liquid;
3) Uniformly stirring water and straws in a ratio of 1.3.
The beneficial effects of the present invention are further illustrated by the following specific experimental examples:
1 solid fermentation of pepper straw
Pulverizing Capsici fructus straw into 2cm, recovering Bacillus subtilis strain, and enlarging culture to obtain powder with bacterial content of 1 × 10 10 cfu/mL, adding bacterial liquid with the water-straw ratio of 1, stirring, sealing, and fermenting at 25 ℃ for 35 days to obtain the microbial fertilizer.
2 the effect of adding fermented pepper straws into the feed on the feeding effect of the broilers
2.1 test animals
Healthy K9 broiler chickens of 120 feather and 30 days old were purchased from Guiyang Huifeng poultry industries, inc.
2.2 composition of basal diet
The basic ration is corn-soybean meal type, and is prepared according to Chinese chicken feeding standard 2004 (NY/T33-2004), and the composition and the nutrition level of the basic ration are shown in Table 1.
TABLE 1 basal diet ingredient composition and nutritional level
Note: (1) the premix is provided for each kilogram of compound feed: vitamin A2 700IU; vitamin D400 IU; 10IU of vitamin E; vitamin K0.5 mg; vitamin B12.007 mg; thiamine 2mg; 5mg of riboflavin; 10mg of pantothenic acid; 30mg of nicotinic acid; pyridoxine 3mg; biotin 0.1mg; folic acid 0.5mg; choline (750 mg); 11.8mg of iron; 0.15mg of copper; 69.4mg of manganese; 56.3mg of zinc; 0.7mg of iodine; selenium 0.27mg. (2) The nutrient components are calculated values.
2.3 test grouping and handling
The test was conducted at animal hospital, southern school district of Guizhou university, from 10 months 2020 to 11 months 2020.
The 120-feather male K9 broilers are randomly divided into a control group and a test group by 2 groups according to the principle that the initial weight difference is not significant, each group is 3 repetitions, and each repetition is 20 feathers. The control group was fed a basal diet, and the test group was prepared by adding 2% (2% obtained from preliminary experiments) of fermented pepper straw feed to the broiler basal diet.
2.4 Breeding management
The test chicken coop is a closed three-layer metal cage-type coop, and the average feeding density is 10 chickens/m 2 The temperature of each cage was varied by. + -. 2 ℃ during the forward test period of 28 days. Feeding at 8, 12, 00 and 19; illumination is carried out for 24 hours; the test chickens were fed and immunized according to the conventional immunization program.
2.5 sample Collection
After the feeding test is finished and the chicken is fasted (without water prohibition) for 12 hours, randomly drawing 6 test chickens for each repeated group, weighing the test chickens, slaughtering the test chickens after bloodletting by jugular veins, taking slaughtered blood by using test tubes, standing the slaughtered blood in a water bath at 37 ℃, centrifuging the blood for 15 minutes at 3 000r/min at 4 ℃, taking supernate, subpackaging the supernate in 1.5mL EP tubes, and storing the EP tubes in a low-temperature refrigerator at 20 ℃ below zero for later analysis and detection of blood biochemical indexes.
Dissecting spleen, bursa of fabricius and thymus of test chicken after serum collection, sucking off water on the surface of organs by using filter paper, weighing and recording by using an electronic balance, taking out the cecum content of the chicken, putting the cecum content into an EP tube, and storing in a low-temperature refrigerator at-80 ℃.
2.6 influence of fermented Capsici fructus stalks on broiler feeding effect
2.6.1 Effect of fermented pepper straw added into feed on broiler chicken production performance
2.6.1.1 measurement indexes and methods
The Initial Weight (IW), average Daily Gain (ADG), average Daily Feed Intake (ADFI) and feed weight ratio (F/G) were recorded for the test chickens.
(1) The final weight is: the weight of the living body of the test chicken after fasting for 12 hours before slaughter was directly weighed by an electronic scale, and the unit thereof was expressed in kilograms (kg).
(2) Daily Gain (ADG): daily gain = (end-initial weight) × 1000/day, units are in grams/day (g/d).
(3) Average Daily Feed Intake (ADFI): average daily feed intake = total feed consumed/number of test days
(4) Material weight ratio (F/G): feed weight ratio = average daily feed intake/average daily gain
2.6.1.2, see Table 2.
TABLE 2 influence of fermented pepper straw addition to feed on broiler productivity
Note: the difference is significant (P < 0.05) as indicated by the different lower case letters in the table.
As a result: as can be seen from Table 2, compared with the control group, the addition of the pepper straws to the diet has no significant influence on the feed weight ratio and the average daily feed intake of the broiler chickens (P is more than 0.05), and the daily gain of the broiler chickens is significantly increased (P is less than 0.05).
2.6.2 Effect of fermented pepper straw added into feed on broiler chicken immune function
2.6.2.1 Immunity index
Immune organ index = immune organ weight (g)/body weight (kg).
Immunoglobulins A (IgA), M (IgM), G (IgG) in serum. The assay was performed with a microplate reader according to the kit instructions, and the kit was purchased from Kexin BioCorp, guizhou.
2.6.2.2 the influence on the immune organ index of broilers, the results are shown in Table 3.
TABLE 3 influence of the addition of pepper straw in the feed on the immune system of broilers
Note: the different lower case letters in the table indicate significant differences (P < 0.05).
As a result: as can be seen from Table 3, the bursal index, thymus index and spleen of the broiler in the test group were significantly increased by 180.95%, 70.82% and 27.77%, respectively (P < 0.05), compared to the control group, indicating that the effect on bursa of Fabricius is greater than that of spleen and thymus after fermented pepper straw was added to the diet.
2.6.2.3 on broiler immunoglobulin, results are shown in table 4.
TABLE 4 influence of fermented pepper straw added in feed on broiler serum immunity index
Note: the difference is significant (P < 0.05) as indicated by the different lower case letters in the table.
As a result: as can be seen from table 4, igG in serum of the treated group was increased by 39.49% (P < 0.05) compared to the control group; there was no significant difference in serum IgA, and IgM content. The fact that the addition of the pepper straws to the daily ration mainly has great influence on serum immunoglobulin IgG is shown.
2.6.3 Effect of adding fermented pepper straw into feed on broiler serum antioxidant index
2.6.3.1 Oxidation resistance index
The determination of the total antioxidant capacity (T-AOC) of serum, glutathione peroxidase (GSH-PX), superoxide dismutase (SOD), peroxidase (POD), malondialdehyde (MDA) and Catalase (CAT) adopts a kit provided by Kyoto biological company in Guizhou province.
2.6.3.2, see Table 5.
TABLE 5 influence of fermented pepper straw added in the feed on the oxidation resistance of broiler serum
Note: the difference is significant (P < 0.05) as indicated by the different lower case letters in the table.
As a result: the influence of the fermented pepper straw feed on the serum antioxidant effect of the broiler is shown in table 5, the content of GSH-PX is remarkably improved by 32.27% (P < 0.05) compared with that of a control group, the content of MDA is reduced by 22.27%, and the difference is remarkable (P < 0.05).
2.6.4 Effect of fermented pepper straw added into feed on broiler cecal microorganisms
2.6.4.1 detection Standard
Flash 1.2.7 software was used for screening and quality control of microbial sequencing raw data, sequencing data were combined and sorted to an operational classification unit (OTU) based on 97% sequence similarity, and alpha diversity of sample communities was analyzed by analyzing Chao1 index, ace index, shannon and Simpson diversity index, which reflect community richness. The QIIME 1.8.0 software is utilized to obtain the composition and abundance distribution table of each sample on the classification level of the phylum and the genus, the abundance difference between the phylum and the genus is compared, the Beta diversity analysis is carried out by utilizing the QIIME software to compare the diversity of the species, the principal component analysis (NMDS) is carried out on the community composition of the genus, and the analysis of the characteristic components of different samples can reflect the difference and the distance between the samples.
2.6.4.2 cecal microbiological sequencing quality analysis and alpha diversity estimation
The dilution curve can be used to determine whether the amount of sequencing per sample is sufficient. The sharp rise in the curve indicates insufficient sequencing and an increase in the number of sequences is required. Conversely, it indicates that the sample sequence is sufficient for data analysis and comparison. See fig. 1, 2.
As a result: the results of the cecal dilution curves show that as the amount of sequencing increases for each treatment group, the OUT number gradually saturates (the curves flatten OUT), indicating that the depth of sequencing is sufficient to cover all species in the sample.
2.6.4.3 estimation of caecal microbial alpha diversity
The α diversity analysis reflects the diversity and abundance of microbial communities, with Shannon index and Simpson index assessing the community diversity; the Chao1 index and ACE index assess flora abundance. The larger the Shannon diversity index, the higher the community diversity, see Table 6.
TABLE 6 influence of the addition of fermented Capsici fructus stalks in the feed on the broiler cecal alpha diversity index
As a result: as can be seen from table 6, the cecal α diversity index shows no significant difference in Shannon index, simpson index, ACE and Chao1 index between the control group and the treatment group (P > 0.05); in addition, the coverage rate of all samples of the caecum reaches 0.999, which shows that the sequencing result has good accuracy and repeatability.
2.6.4.4 Effect of fermented pepper straw added into feed on broiler cecal microbial flora
Through classification and identification of broiler cecal phyla and genus level sequences, the cecal dominant bacteria phyla of the control group and the treatment group are bacteroides (48.98%, 45.85%) and Firmicutes
(39.34%, 42.85%), proteobacteria (3.69%, 4.03%), actinobacterium (2.26%, 2.86%), on the genus level, the dominant genus cecal of the control and treatment groups were bacterioides (28.37%, 23.23%), alistipes (5.80%, 7.37%), uncultured _ bacterium _ f _ Murizobacteriaceae (5.19%, 7.34%) ruminococcus upaeae ucg-014 (4.94%, 4.88%).
2.6.4.5 differences in bacterial Community composition
At the level of the cecum gate (see table 7), the relative abundance of firmiciutes in the treated group was not significant, although higher than that in the control group, whereas the relative abundances of bacteroides, tenericates and elsusimicrobia were significantly lower than that in the control group (P < 0.05). At the cecal level (see table 7), the relative abundance of Bacteroides in the test group was significantly lower than that of the control group (P < 0.05) and the relative abundance of Faecalibacterium was significantly higher than that of the control group (P < 0.05), with a decrease in the relative abundance of Lactobacillus of 37.46% compared to the control group, but with a significant difference (P > 0.05). The number of taxa with significant differences at the phylum and genus levels was 2, 23 (Table 3.7), indicating differences in colony composition between the treated and control groups (P < 0.05).
2.6.4.6 differences in bacterial Community composition
At the level of the cecum gate (table 7), the relative abundance of firmiciutes in the treated group was higher than that in the control group but not significant, whereas the relative abundances of bacteroides, tenericates and elsusimrobia were significantly lower than that in the control group (P < 0.05). At the cecal level (see table 7), the relative abundance of Bacteroides in the test group was significantly lower than that of the control group (P < 0.05) and the relative abundance of Faecalibacterium was significantly higher than that of the control group (P < 0.05), with a 37.46% reduction in Lactobacillus relative abundance compared to the control group, but with a significant difference (P > 0.05). The number of taxa with significant differences at the phylum and genus levels was 2, 23 (see table 8), indicating differences in colony composition between the treated and control groups (P < 0.05).
TABLE 7 influence of the addition of fermented Capsici fructus stalks to the feed on the relative abundance of the chicken cecal phylum and genera
TABLE 8 statistical Table of Metastats pairwise comparison test results between samples (groups)
2.6.4.7 Effect of adding fermented pepper straw into feed on Beta diversity of broiler chicken biological group
Non-metric multidimensional calibration (NMDS) is a non-linear model that better reflects the non-linear structure of the ecological data. Based on the species information contained in the samples, the NMDS analysis reflects the multidimensional space in the form of points, while the degree of difference between different samples is reflected by the distance between points, which may reflect the difference between sample groups and within groups. The results of NMDS analysis based on OTU level are shown in fig. 3, and it can be seen that there is a certain difference in the biological communities of the control group and the experimental group.
2.7 data processing and analysis
All data are subjected to Excel 2010 primary processing, statistical analysis of chicken production performance, immune organ indexes, serum antioxidant indexes and serum immune index data is carried out by using test data and SPSS 19.0 software, single-factor variance analysis is used for carrying out difference significance test, when the difference is significant, a Duncan's method is used for carrying out multiple comparisons, the results are all expressed by' mean value +/-standard deviation ', and P < 0.05' is used as a difference significance judgment standard.
2.8 conclusions and problems
(1) The daily gain of the broiler chicken is obviously improved by adding the fermented pepper straws into the feed, and the bursal index, the thymus index and the spleen of the broiler chicken are respectively improved by 180.95 percent, 70.82 percent and 27.77 percent. The serum immunoglobulin IgG has larger influence, and is increased by 39.49 percent compared with a control group; the content of glutathione peroxidase is obviously improved by 32.27 percent compared with the content of a control group, and the content of malondialdehyde is obviously reduced by 22.27 percent.
(2) Classifying and identifying broiler cecal phyla and genus horizontal sequences, wherein dominant phyla of cecal is bacteroidetes and firmicutes; the dominant bacteria are Bacteroides and Clostridium molesta. The relative abundance of the phylum of the treatment group, i.e., the phylum of the traceobacter and the phylum of the cyanobacteria, is obviously lower than that of the control group at the level of the phylum of the cecum. The relative abundance of Bacteroides is obviously reduced at the cecal level compared with that of a control group, and the relative abundance of coprococcus is obviously improved.
(3) The influence of the fermented pepper straws added into the feed on the production performance of the broilers is as follows: the pepper straw has high fiber content and serious lignification, and has low feeding value in the production of monogastric animals. However, with the continuous development and popularization of the biological fermentation technology, the feeding value of the pepper straws can be improved by fermenting the pepper straws through microorganisms, and the method is a new direction for the development of the breeding industry. Because the growth cycle and the digestive system of the chicken are short, the chicken can meet the growth requirement in a short time, and the predigestion effect of the fermented feed can improve the quality of the feed, promote the digestion and absorption of the feed by animals (Chenting and the like, 2019) and promote the growth and development of the chicken. Europe and the other people utilize bacillus subtilis solid state fermentation rice bran to mix into daily ration and feed Linwu duck, and researches find that the average daily gain of Linwu duck can be effectively improved by adding bacillus subtilis into the daily ration, and the feed conversion ratio and the death rate are reduced. The Qinqui (2020) adds 200g/t of bacillus subtilis in the daily ration, so that the average daily gain of 22-42d broilers is obviously increased, and the average daily gain and the average daily feed intake of the broilers in 1-21 days are obviously reduced. The experimental result shows that the average daily gain can be obviously increased and the feed-weight ratio can be reduced by properly adding 2% of fermented straw feed into the daily ration of the broiler in the growing period, but the average daily feed intake is not obviously affected. Part of the reasons may be that bacillus subtilis exposes hemicellulose-xylan chains and lignin polymer ester bonds of straws for enzymolysis in the straw fermentation process, and (senecine and the like, 2020) decomposes partial macromolecular proteins, complex polysaccharides, fats, proteins and the like into small molecular substances, so that the palatability of the feed is improved, and the digestion utilization rate of nutrient substances and the growth performance of animals are improved. The other part is related to factors such as phytase, protease, etc. produced during the fermentation process of bacillus subtilis, and intestinal micro-ecology stability by biological oxygen deprivation in intestinal tract (sunlin 2015).
(4) The influence of the fermented pepper straws added into the feed on the immune function of the broiler chicken is as follows: thymus, spleen and bursa of fabricius are important immune organs of broiler chickens, are the main sites for the formation and differentiation of various immune cells in the body, participate in cellular and humoral immunity of the body, produce a large amount of antibodies to protect the body from the invasion of foreign pathogens, and the weight of immune organs determines the strength of immune functions, the larger the absolute weight and relative weight of which is, the stronger the cellular and humoral immune functions of the body (treemark, 2010; songi, 2007. In the test, the fermented pepper straws are added into the daily ration of the broiler chicken, so that the thymus index, the spleen index and the bursal disease index can be obviously improved, wherein the bursal disease index has the most obvious effect and is improved by 180.95 percent compared with a blank group. The bacillus subtilis fermented pepper straws can obviously improve the immunity level of the broiler chickens, so that the immunity of the broiler chickens to diseases is improved, and the research result is similar to that of the Rotenqin (2020).
Immunoglobulin is one of the most critical substances in the immune system of animals, has important immune and physiological regulation effects, can generate specific combination with corresponding antigens so as to realize various immune effects, and the content of the immunoglobulin in serum reflects the level of the immune function of the animal body (Zhang Xiao Rong et al, 2014; guaifang et al, 2004). The experiment shows that the addition of the fermented straws into the basic feed can improve the level of IgG in the serum of the broiler chicken by 39.49 percent compared with a control group, and the content difference of IgA and IgM in the serum is not obvious. The result shows that the addition of the pepper straws in the diet has great influence on serum immunoglobulin IgG, but has no obvious influence on other nonspecific immunity indexes such as immunoglobulin, cell factors and the like. The reason for the analysis may be that the bacterial components of bacillus subtilis are used as antigens to stimulate intestinal tracts or act on immune organs of animals in the form of immune adjuvants, so that the bacillus subtilis proliferates in digestive tracts, continuously synthesizes various beneficial substances such as vitamins, amino acids and the like, and promotes the growth and development of the immune organs (Yudongfeng et al, 2010).
(5) The effect of adding fermented pepper straws into the feed on the oxidation resistance index of the broiler serum is as follows: the antioxidant system of living cells comprises three main lines of defense: the first one consists of superoxide dismutase, glutathione peroxidase, catalase, antioxidant substance and the like and is responsible for inhibiting the generation of free radicals and lipid peroxidation. By measuring the contents of SOD and glutathione peroxidase, the damage degree of free radicals to organisms can be known. In the test, the contents of T-AOC, SOD, POD and CAT in the broiler fed with the straw feed tend to be increased compared with those in a control group, but the differences are not obvious (P is more than 0.05); the content of GSH-PX is obviously improved by 32.27 percent (P < 0.05) compared with the control group, and the content of malondialdehyde is obviously reduced by 22.27 percent (P < 0.05). The method shows that the addition of the fermented pepper straws into the daily ration of the broiler chicken can improve the activities of the antioxidant enzymes SOD and GSH-PX of the broiler chicken, further improve the total antioxidant capacity (T-AOC) of the organism and reduce the activity of malonaldehyde. The antioxidant property of the pepper straw fermented by the bacillus subtilis and the antioxidant mechanism in vivo still need to be further and deeply researched.
(6) The influence of the fermented pepper straws added into the feed on the cecal microorganisms of the broilers is shown as follows: normally, the microbial flora in the intestinal tract of animals keeps a microecological balance, and the balance of the microbial flora in the intestinal tract plays a key role in the digestion and absorption of nutrient substances and the maintenance of the health of a host. The research shows that the difference between the sequencing quantity of the broiler caecal flora after the bacillus subtilis is fed to ferment the pepper straws and the control group is not obvious, which shows that the broiler caecal flora microorganism species has no obvious effect when the bacillus subtilis is fed to ferment the pepper straws. The cecal alpha diversity index in this trial showed no significant difference in Shannon index, simpson index, ACE and Chao1 index between control and treatment groups (P > 0.05). The method shows that the fermented pepper straws added into the broiler feed have no influence on the diversity and abundance of intestinal flora of broilers.
Researches in recent years find that the caecum is the part containing a large number of microorganisms with the highest concentration in the intestinal tracts of livestock and poultry. The test detects the graduation and diversity change of the flora in the cecum under the regulation and control of the bacillus subtilis fermented pepper straw daily ration by a bacteria 16S-rDNA high-throughput sequencing technology, and the result shows that bacteria of Bacteroides (Bacteroidetes), firmicutes (Firmicutes), proteobacteria (Proteobacteria) and Actinobacteria (Actinomycetes) are dominant, and the result is the same as the result of researching the types of the microorganisms in pig manure and intestinal tracts by utilizing high-throughput sequencing by Park (2014) and Roc (2015). Yanlina (2014) studies also found that the cecal microorganisms in the intestinal tract of monogastric animals have high diversity, wherein the dominant flora is firmicutes and bacteroidetes. The results showed that bacteria of the phylum firmicutes, proteobacteria, actinomycetes and bacteroidetes predominated, which is the same as that of Park (2014) which studies the species of microorganisms in pig faeces and in the intestine using high throughput sequencing. Bacteroidetes and firmicutes share a mutually promoting symbiotic relationship, and they together promote the host to absorb or store energy, bacteroidetes decreases or firmicutes increases or bacteroidetes/firmicutes ratio decreases, contributing to fat absorption accumulation (guo junlan, 2009 turnbaugh P J et al, 2006 ley R E et al, 2005 ley R E et al, 2006), bacteroidetes decreases in the test group, firmicutes, proteobacteria and actinomycetes are slightly higher than the control group, and all have a tendency to increase. The test finds that the absolute dominant flora in the cecum of the broiler chicken is bacteroidetes, the proteobacteria flora is less in distribution, and after the fermentation product of pepper straws fermented by adding bacillus subtilis is fed, the abundance of microorganism species is increased, so that the cyanophyta, the tracegate and the like are obviously increased.
At the genus level, bacteroides are normal flora abundantly present in humans and animals, clostridium tenella is a major member of the gut symbiotic firmicutes, the most common bacteria in the human gut, and one of the most predominant butyrate-producing bacteria, while butyric acid can provide energy to the gut and exert an anti-inflammatory effect (Barcenilla et al,2000 louisp et al, 2009. Studies have shown that Clostridium tenella has anti-inflammatory effects and is beneficial to the host, while Bacteroides has pro-inflammatory effects and is harmful to the host. Some types of bacteroides can also produce enterotoxins, inducing the development of colon cancer (Kelly et al, 2004). In the test, the relative abundance of broiler cecal bacteroides is obviously reduced compared with that of a control group, and the relative abundance of tender clostridium is obviously higher than that of the control group, so that the bacillus subtilis is fed to ferment the pepper straws, the body inflammation can be reduced, the intestinal flora structure of broilers is improved, the intestinal disease cause is prevented, the intestinal health is favorably maintained, and the growth of the broilers is promoted. Studies have shown that bacteroidetes groups are beneficial for the normal development of the gastrointestinal tract and that they produce butyrate, which is the major product of colonic fermentation and has anti-tumor and gut health-preserving properties.
Although the invention has been described in detail hereinabove by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that modifications and improvements can be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (5)
1. The feed containing the fermented pepper straw is characterized by comprising a basic ration and fermented pepper straw powder, wherein the basic ration comprises the following components in parts by weight: 66.9 parts of corn, 24 parts of soybean meal, 3.5 parts of soybean oil, 0.8 part of stone powder, 0.3 part of salt, 0.5 part of calcium hydrophosphate and 4 parts of premix; the fermented pepper straw powder accounts for 2 percent of the basic ration in parts by weight;
the preparation method of the fermented chili straw powder comprises the following steps:
1) Smashing pepper straws to 2cm in length;
2) Resuscitating and expanding the bacillus subtilis strain to prepare a bacterial liquid with the bacterial quantity more than or equal to 1 multiplied by 1010cfu/m L;
3) Uniformly stirring water and straws in a ratio of 1.
2. The feed as claimed in claim 1, wherein said pepper straw is used in animal husbandry.
3. The feed as claimed in claim 1, wherein the fermented pepper straw powder is used for raising broiler chickens.
4. The feed of claim 3, wherein the fermented capsicum straw powder is applied to the aspects of broiler productivity, immune function, serum antioxidant index and cecal microorganisms.
5. The feed of claim 1, wherein the fermented pepper straw powder has a mellow taste or an acid taste.
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| CN1663422A (en) * | 2005-04-19 | 2005-09-07 | 单德章 | Zymophyte of plant straw feedstuff and its preparation |
| CN104381708A (en) * | 2014-12-10 | 2015-03-04 | 青岛德润电池材料有限公司 | Chicken feed capable of improving immunity and preparation method of chicken feed |
| CN105533133A (en) * | 2015-12-04 | 2016-05-04 | 重庆三零三科技有限公司 | Method of preparing feed from stem and leaf of pepper |
| CN105941849A (en) * | 2016-06-12 | 2016-09-21 | 淮北嘉吉农牧科技有限公司 | Rumen fermentation promoting pig feed |
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| CN103549125B (en) * | 2013-09-22 | 2015-12-02 | 安徽兴牧畜禽有限公司 | A kind of chicken feed improving resistance against diseases |
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| CN104381708A (en) * | 2014-12-10 | 2015-03-04 | 青岛德润电池材料有限公司 | Chicken feed capable of improving immunity and preparation method of chicken feed |
| CN105533133A (en) * | 2015-12-04 | 2016-05-04 | 重庆三零三科技有限公司 | Method of preparing feed from stem and leaf of pepper |
| CN105941849A (en) * | 2016-06-12 | 2016-09-21 | 淮北嘉吉农牧科技有限公司 | Rumen fermentation promoting pig feed |
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