CN114009580B - Broussonetia papyrifera fermentation total mixed ration formula and processing technology and application thereof - Google Patents
Broussonetia papyrifera fermentation total mixed ration formula and processing technology and application thereof Download PDFInfo
- Publication number
- CN114009580B CN114009580B CN202111234873.5A CN202111234873A CN114009580B CN 114009580 B CN114009580 B CN 114009580B CN 202111234873 A CN202111234873 A CN 202111234873A CN 114009580 B CN114009580 B CN 114009580B
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- China
- Prior art keywords
- total mixed
- paper mulberry
- microbial inoculum
- fermentation
- crawler belt
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Classifications
-
- 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
-
- 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
-
- 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
-
- 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
- A23K10/38—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 from distillers' or brewers' waste
-
- 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
-
- 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
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K40/00—Shaping or working-up of animal feeding-stuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- 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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Health & Medical Sciences (AREA)
- Mycology (AREA)
- Botany (AREA)
- Inorganic Chemistry (AREA)
- Sustainable Development (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Fodder In General (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a paper mulberry type fermentation total mixed ration formula and a processing technology and application thereof, neutral washing fiber/non-fibrous carbohydrate (non-fiber carbohydrates/Neutral detergent fiber, NFC/NDF) values and the addition level of a composite bacterial agent of paper mulberry type fermentation total mixed ration (Fermentation total mixed ration, FTMR) are taken as test factors, the pH value, organic acid and mycotoxin content of each treatment group are measured by adopting a 2-factor cross grouping test design, and then the paper mulberry type fermentation total mixed ration is explored through data statistics and analysis.
Description
Technical Field
The invention relates to the field of feed invention, in particular to a paper mulberry type fermented total mixed ration formula, a processing technology and application thereof.
Background
At present, the shortage of protein source feed is still a short plate for restricting the high-quality development of livestock and feed industry in China, and the development strength of non-conventional protein feed resources such as by-products in agricultural product processing, feed plants, residues and the like is increased, so that one of effective strategies is realized [1] . Broussonetia papyrifera (Broussonetia papyrifera), also known as Broussonetia papyrifera, broussonetia papyrifera,Deer young tree and the like, perennial deciduous tree is Moraceae (Moraceae), broussonetia (Broussonetia) deciduous tree plant, is multifunctional rural tree species with long application history in China, and is widely applied to industries such as papermaking, feed, medicine and the like [2] . Many researches show that the content of Crude Protein (CP) in the broussonetia papyrifera leaves reaches about 20%, the nutrient types and contents of amino acids, vitamins, minerals and the like are comprehensively compared, the nutritive value of the broussonetia papyrifera leaves is between that of corn and soybean, and the broussonetia papyrifera leaves is a high-quality protein source Crude feed, and has good development prospect in a suitable planting area [3] . In general, the processing and utilizing method after the paper mulberry is timely mowed comprises cutting, direct silage, drying and pulverizing, simple silage, biological fermentation and the like, wherein the biological fermentation method is one of the focus points of the current research due to the effects of improving palatability, degrading fiber content, improving nutrient digestibility and the like. At present, the fermented paper mulberry feed is widely applied to piglets [4] Fattening pig [5] Milk cow [6] Beef cattle [7] And broiler chicken [8] The method has good effect in the daily ration, however, most reports are focused on the application research of the paper mulberry type fermented feed finished product, and the system research of reasonably preparing the production end feed is lacking.
Research shows that the quality of the fermented feed is mainly determined by the comprehensive factors of fermentation technology (mixing uniformity, compactness, tightness and the like), raw material composition structure, fermentation strain type, addition level and the like [10] . In addition, it is reported that neutral washing fiber (Neutral detergent fiber, NDF) and non-fibrous carbohydrate (non-fiber carbohydrates, NFC) in ruminant ration are respectively used as slow and fast degradation carbohydrate components of rumen, and the relative content of the neutral washing fiber and the non-fibrous carbohydrate can influence characteristic indexes of feed intake and production performance by regulating and controlling the number of rumen times, rumen fermentation characteristics and filling degree and secretion and release of hormone in alimentary canal [10-12] . The NDF/NFC is used as a characterization index for evaluating the structural rationality of the ration formula by the prior researchers [13-15] The fermentation complete mixing day (Fermentation total mixed ration, FTMR) is therefore of practical significance in terms of formulation considering NDF/NFC.
The invention discloses a method for feeding Hehuan cattle with silage, a concentrated feed and compound fungus chaff compound feed, and discloses daily ration comprising silage containing the silage, a concentrate supplement and the compound fungus chaff feed. The ration weight ratio is 0-84 parts of paper mulberry silage, 0-84 parts of silage corn, 17-30 parts of concentrate supplement and 0.5-5 parts of compound fungus chaff feed. The formula of the daily ration compound fungus chaff feed comprises the following components: 1-10 parts of ganoderma lucidum fruiting body powder, 30-60 parts of ganoderma lucidum fungus chaff powder, 10-30 parts of rosemary hay powder, 10-30 parts of evening primrose hay powder and 1-5 parts of eucommia ulmoides leaf powder. The method fully utilizes local Chinese herbal medicine resources, establishes Chinese herbal medicine health-care formula feed, is applied to healthy and efficient breeding of local beef cattle, evaluates feed value from breeding effect and meat quality, establishes a local beef production method, and promotes breeding effect improvement.
Technical problem of application number CN 202010703373.0: 1) The fermentation time is long, which is not beneficial to industrial production; 2) The feed has low protein, lactic acid and propionic acid content, and is not easy to digest and absorb after eating by cattle. 3) The feed has poor stability and is not easy to store.
Therefore, the team of the invention determines the pH value, the organic acid and the mycotoxin content of each treatment group according to the formula of the prior study (application number CN202010703373.0, the method for feeding the Szechwan black cattle with compound fungus chaff compound feed) by taking the carbohydrate structure (NFC/NDF) and the compound fungus addition level as test factors, adopting 2-factor cross grouping test design, and respectively taking the NFC/NDF values (1.08, 1.34, 1.60 and 1.86) and the compound fungus addition level (0, 0.4 and 0.8g/kg DM) of fermentation substrates as 2 factors, and then exploring a complete mixed ration for fermentation of the paper mulberry by data statistics and analysis.
Disclosure of Invention
The invention aims to provide a paper mulberry type fermented total mixed ration.
The invention further aims to provide a processing technology of the paper mulberry type fermented total mixed ration.
The invention further aims to provide an application of the fermentation substrate NFC/NDF value and the composite microbial inoculum in the paper mulberry type fermentation total mixed ration.
The broussonetia papyrifera fermentation total mixed ration consists of raw materials, concentrate and a composite microbial inoculum, wherein the raw materials comprise 30-50 parts of corn silage and 20-60 parts of whole broussonetia papyrifera; the fine material comprises 7-21 parts of corn, 1.7-4 parts of wheat bran, 0.9-5 parts of bean pulp, 0.5-1 part of corn dehydrated alcohol lees, 0.1-0.4 part of calcium carbonate, 0.08-0.3 part of calcium hydrophosphate, 0.04-0.2 part of sodium bicarbonate, 0.08-0.3 part of salt and 0.03-0.15 part of premix; the addition amount of the composite microbial inoculum is 3-5g/kg.
Preferably, the method comprises the steps of,
the broussonetia papyrifera fermentation total mixed ration consists of 42.47 parts of corn silage and 34.75 parts of whole broussonetia papyrifera; the concentrate comprises 15.50 parts of corn, 2.42 parts of wheat bran, 3.35 parts of soybean meal, 0.68 part of corn dehydrated alcohol lees, 0.27 part of calcium carbonate, 0.18 part of calcium hydrophosphate, 0.11 part of sodium bicarbonate, 0.18 part of salt and 0.08 part of premix; the addition amount of the composite microbial inoculum is 4g/kg.
The processing technology of the paper mulberry type fermented total mixed ration comprises the following steps:
1) Starting a transmission crawler belt, respectively conveying the whole broussonetia papyrifera and the corn silage with the water content of 60-80% to a pulverizer, pulverizing to 2-3cm, and conveying to a mixing stirrer through an automatic weighing machine and the crawler belt;
2) The water content is divided into 12-15% concentrate, and the concentrate is conveyed into a mixing stirrer through an automatic weighing device and a crawler belt;
3) Taking a formula amount of the composite microbial inoculum in a microbial inoculum sprayer, and spraying the composite microbial inoculum into a mixing stirrer;
4) Starting a mixing stirrer, stirring for 20-40min, and conveying to a pressing bucket through an inductor and a crawler belt for compaction treatment;
5) And (3) conveying the compacted daily ration to a fermentation chamber through an automatic weighing device and a crawler belt, and fermenting for 20-40 days under the condition of room temperature and humidity kept at 40-50%, so as to obtain the paper mulberry type fermented total mixed daily ration.
Preferably, the method comprises the steps of,
the processing technology of the paper mulberry type fermented total mixed ration comprises the following steps:
1) Starting a transmission crawler belt, respectively conveying the whole broussonetia papyrifera and the corn silage with the water content of 70% into a pulverizer, pulverizing to 2.5cm, and conveying the whole broussonetia papyrifera and the corn silage into a mixing stirrer through an automatic weighing machine and the crawler belt;
2) The refined materials with the water content of 13 percent are conveyed into a mixing stirrer through an automatic weighing device and a crawler belt;
3) Taking a formula amount of the composite microbial inoculum in a microbial inoculum sprayer, and spraying the composite microbial inoculum into a mixing stirrer;
4) Starting a mixing stirrer, stirring for 30min, and conveying to a pressing bucket through an inductor and a crawler belt for compaction treatment;
5) And (3) conveying the compacted daily ration to a fermentation room through an automatic weighing device and a crawler belt, and fermenting for 30 days under the condition of room temperature and humidity kept at 45%, so as to obtain the paper mulberry type fermented total mixed daily ration.
The invention relates to application of a fermentation substrate NFC/NDF value and a composite microbial inoculum of a paper mulberry type fermentation total mixed ration in the paper mulberry type fermentation total mixed ration.
The NFC/NDF value of the fermentation substrate is 1.08-1.86, the composite microbial inoculum is lactobacillus plantarum and saccharomyces cerevisiae, and the addition amount is 2-10g/kg DM.
Preferably, the method comprises the steps of,
the NFC/NDF value of the fermentation substrate is 1.34-1.60, the composite microbial inoculum is lactobacillus plantarum and saccharomyces cerevisiae, and the addition amount is 4-8g/kg DM.
It is further preferred that the composition of the present invention,
the NFC/NDF value of the fermentation substrate is 1.60, the composite microbial inoculum is lactobacillus plantarum and saccharomyces cerevisiae, and the addition amount is 4g/kg DM.
The application of the invention is the application in the fermentation quality and mycotoxin content of the broussonetia papyrifera fermentation total mixed ration.
The fermentation quality is the NFC/NDF value, the pH value and the organic acid content of a fermentation substrate; the mycotoxin content is aflatoxin, vomitoxin and zearalenone content.
The parts by weight of the present invention may be conventional units in the art, and may be mg, g, kg, etc.
Advantageous effects
1. According to the invention, through the influence of neutral washing fiber/non-fibrous carbohydrate (non-fiber carbohydrates/Neutral detergent fiber, NFC/NDF) values and the addition level of a compound bacterial agent on the fermentation quality and mycotoxin content of the fermentation total mixed ration (Fermentation total mixed ration, FTMR), 2 factors are respectively adopted to determine the pH value, the organic acid and the mycotoxin content of each treatment group by adopting a 2-factor cross grouping test design, and the NFC/NDF values (1.08, 1.34, 1.60 and 1.86) and the compound bacterial addition level (0, 0.4 and 0.8g/kg DM) of a fermentation substrate, and then the data statistics and analysis are carried out, so that the paper mulberry type fermentation total mixed ration is researched, the mixed ration has short fermentation time, the pH value is lower than 4.20, the storage stability, the protein, the lactic acid and the propionic acid content are high, the digestion and the absorption of livestock are facilitated, and the reference is provided for developing paper mulberry fermentation feed products.
2. The invention examines the pH value and the organic acid content of the fermented total mixed ration: the results show that: on the one hand, under the condition of different carbohydrate structures (NFC/NDF values), the pH value of the fermentation substrate is lower than 4.20, which indicates the substrate structure rationality of each treatment group; on the other hand, with the increase of the NFC/NDF value of the diet, the pH value is in a trend of secondarily decreasing integrally, so that the propagation and growth of the diet are promoted, the fermentation process is accelerated, and the fermentation time is shortened.
3. According to the invention, the mycotoxin content of the fermented total mixed ration is examined, and the results show that 3 types of aflatoxin B1, vomitoxin and zearalenone are respectively 8.36-16.33ng/kg, 417-46ng/kg and 83.15-374.05ng/kg, which are lower than limit standards specified by the feed sanitation standards in China (aflatoxin B1 is less than or equal to 20, vomitoxin is less than or equal to 3000 and zearalenone is less than or equal to 500 in other compound feeds), so that the anaerobic conditions created by the test are more suitable. In addition, the carbohydrate structure of the fermentation substrate obviously influences the mycotoxin content in 3, the interaction of the addition amount of the composite microbial inoculum and the 2 factors has obvious influence on the aflatoxin B1 and zearalenone content, and the 2 test factors designed in the research show that the 2 test factors are important factors for regulating and controlling the mycotoxin production, and when NFC/NDF=1.60 of the fermentation substrate, the addition amount of the composite microbial inoculum is 4 and 8g/kg DM mycotoxin content is the lowest, so that the method has great guiding significance on actual production.
4. The invention analyzes the relativity of mycotoxin, pH value and organic acid content of the broussonetia papyrifera fermented feed, finds that the pH value of a fermentation substrate is extremely obviously positively correlated with 3 mycotoxin contents, the lactic acid is extremely obviously negatively correlated, the acetic acid and the propionic acid are extremely obviously or obviously negatively correlated with aflatoxin B1, the butyric acid is extremely obviously positively correlated with aflatoxin B1 and zearalenone, and the result accords with the theory reported by most of previous people: the higher the organic acid content in the fermented feed in the anaerobic state, the stronger the growth inhibition capability of harmful mould is, so that the organic acid content in the actual production can reflect the mould growth state of the fermented feed to a certain extent, and can be used as an important evaluation index of fermentation quality.
Detailed Description
The following examples are illustrative of the invention and are not intended to limit the scope of the invention.
Example 1 formulation
Raw materials are 42.47kg of corn silage and 34.75kg of whole broussonetia papyrifera; the concentrate comprises 15.50kg of corn, 2.42kg of wheat bran, 3.35kg of bean pulp, 0.68kg of corn dehydrated alcohol lees, 0.27kg of calcium carbonate, 0.18kg of calcium hydrophosphate, 0.11kg of sodium bicarbonate, 0.18kg of salt, 0.08kg of premix and 4g/kg of compound bacteria.
Example 2 formulation
Raw materials are 31.21kg of corn silage and 57.97kg of whole broussonetia papyrifera; the concentrate comprises 7.14kg of corn, 1.77kg of wheat bran, 0.97kg of bean pulp, 0.54kg of corn dehydrated alcohol lees, 0.13kg of calcium carbonate, 0.09kg of calcium hydrophosphate, 0.05kg of sodium bicarbonate, 0.09kg of salt, 0.04kg of premix and 4g/kg of compound bacteria.
Example 3 formulation
44.80kg of corn silage and 24.12kg of whole broussonetia papyrifera; the concentrate comprises 20.6kg of corn, 3.95kg of wheat bran, 4.46kg of bean pulp, 0.93kg of corn dehydrated alcohol lees, 0.37kg of calcium carbonate, 0.25kg of calcium hydrophosphate, 0.16kg of sodium bicarbonate, 0.25kg of salt, 0.11kg of premix and 4g/kg of compound bacteria.
The formulations of examples 1-3 were prepared according to the preparation methods of examples 4-6, respectively.
Example 4 preparation method
1) Starting a transmission crawler belt, respectively conveying the whole broussonetia papyrifera and the corn silage with the water content of 70% into a pulverizer, pulverizing to 3cm, and conveying the crushed broussonetia papyrifera and the corn silage into a mixing stirrer through an automatic weighing device and the crawler belt;
2) The refined materials with the water content of 13 percent are conveyed into a mixing stirrer through an automatic weighing device and a crawler belt;
3) Taking a formula amount of the composite microbial inoculum in a microbial inoculum sprayer, and spraying the composite microbial inoculum into a mixing stirrer;
4) Starting a mixing stirrer, stirring for 30min, and conveying to a pressing bucket through an inductor and a crawler belt for compaction treatment;
5) And (3) conveying the compacted daily ration to a fermentation room through an automatic weighing device and a crawler belt, and fermenting for 30 days under the condition of room temperature and humidity kept at 45%, so as to obtain the paper mulberry type fermented total mixed daily ration.
Example 5 preparation method
1) Starting a transmission crawler belt, respectively conveying the whole broussonetia papyrifera and the corn silage with the water content of 60% into a pulverizer, pulverizing to 2cm, and conveying the crushed broussonetia papyrifera and the corn silage into a mixing stirrer through an automatic weighing device and the crawler belt;
2) The water is divided into 12% concentrate, and the concentrate is conveyed into a mixing stirrer through an automatic weighing device and a crawler belt;
3) Taking a formula amount of the composite microbial inoculum in a microbial inoculum sprayer, and spraying the composite microbial inoculum into a mixing stirrer;
4) Starting a mixing stirrer, stirring for 20min, and conveying to a pressing bucket through an inductor and a crawler belt for compaction treatment;
5) And (3) conveying the compacted daily ration to a fermentation room through an automatic weighing device and a crawler belt, and fermenting for 20 days under the condition of room temperature and humidity kept at 40%, so as to obtain the paper mulberry type fermented total mixed daily ration.
Example 6 preparation method
1) Starting a transmission crawler belt, respectively conveying the whole broussonetia papyrifera and the corn silage with the water content of 80% into a pulverizer, pulverizing to 2.5cm, and conveying the whole broussonetia papyrifera and the corn silage into a mixing stirrer through an automatic weighing machine and the crawler belt;
2) The water content is divided into 15 percent of concentrate, and the concentrate is conveyed into a mixing stirrer through an automatic weighing machine and a crawler belt;
3) Taking a formula amount of the composite microbial inoculum in a microbial inoculum sprayer, and spraying the composite microbial inoculum into a mixing stirrer;
4) Starting a mixing stirrer, stirring for 40min, and conveying to a pressing bucket through an inductor and a crawler belt for compaction treatment;
5) And (3) conveying the compacted daily ration to a fermentation room through an automatic weighing device and a crawler belt, and fermenting for 40 days under the condition of room temperature and humidity kept at 50%, so as to obtain the paper mulberry type fermented total mixed daily ration.
The prepared broussonetia papyrifera fermented total mixed ration of examples 4-6 was tested according to the test method of example 7, respectively.
Example 7 detection method
1) FTMR raw material approximate nutrient determination method: the raw materials were assayed for dry matter, crude protein, crude fat, crude ash, neutral detergent fiber, acid detergent fiber, calcium and phosphorus according to feed analysis and quality control techniques version 2.
2) The PH value measuring method comprises the following steps: 200g of mixed ration sample is put into a juicer, 1800mL of distilled water is added, stirring and mashing are carried out, 4 layers of gauze and qualitative filter paper are used for filtering to obtain leaching liquid, and the pH value of the obtained leaching liquid is immediately measured.
3) Lactic acid, acetic acid, propionic acid and butyric acid content determination: 200g of mixed ration sample is put into a juicer, 1800mL of distilled water is added, stirring and mashing are carried out, 4 layers of gauze and qualitative filter paper are used for filtering to obtain leaching liquid, the leaching liquid is preserved at the temperature of minus 20 ℃, and the measurement is carried out by adopting an Shimadzu LC-20A type high performance liquid chromatograph, and a chromatographic column is adopted: inertSustatin C18, 5 μm by 4.6X250 nm; the mobile phase consists of 0.05mol/L H PO4-KH2PO4 buffer solution with pH of 2.8 and acetonitrile in a volume ratio of 95:5, the flow rate is 0.5mL/min, the column temperature is 16 ℃, the wavelength of an ultraviolet detector is 210nm, and the sample injection volume is 10 mu L.
4) Determination of aflatoxin, vomitoxin and zearalenone contents: 500g of mixed ration sample is taken and is preserved at the temperature of minus 20 ℃, and the contents of aflatoxin, vomit toxin and zearalenone are measured by using an ELISA reagent.
In order to further verify the effectiveness of the present invention, the invention performed a series of verification tests, specifically as follows:
1 materials and methods
1.1 test time and place
The field test of the paper mulberry fermented feed was carried out in the Hao Miao nationality county paper mulberry industry hatching park in the Zaoyi city in the following sense of the city in the year 2020, 7, and the laboratory measurement of the sample was carried out in the institute of grass industry, the agricultural academy of sciences in the Guizhou province.
1.2 test materials
The Broussonetia papyrifera FTMR is a feedable material prepared by chopping whole broussonetia papyrifera and silage corns to 5-8cm, fully mixing with concentrated feed and microbial inoculum according to a certain proportion, compacting, sealing and anaerobic fermentation. The whole broussonetia papyrifera is woody green feed obtained by cutting hybrid broussonetia papyrifera with a height of 1.2m above ground together with stems and leaves of the plants. The viable count of the composite microbial inoculum is Lactobacillus plantarum (Lactobacillus plantarum) > 2.00×10 7 CFU/g, saccharomyces cerevisiae (Saccharomyces cerevisiae) > 2.00×10 7 CFU/g, moisture content is less than or equal to 10%.
1.3 design of experiments
This test was aimed at exploring the effect of neutral wash fiber/non-fibrous carbohydrate (non-fiber carbohydrates/Neutral detergent fiber, NFC/NDF) values and the addition level of the complex inoculant on its fermentation quality and mycotoxin content of broussonetia papyrifera fermentation total mixed ration (Fermentation total mixed ration, FTMR). The test adopts a two-factor cross grouping test design, wherein the two factors are respectively the feed NFC/NDF value and the compound bacteria addition level, and the test design is shown in Table 1. On the basis of the basic formulation, other nutrients remained essentially identical except for the fiber structure and the effective value, and the diet formulation for each NFC/NDF gradient was as shown in table 2 (NFC/NDF values from low to high A, B, C, D, respectively). To ensure that the NFC/NDF values of FTMR follow the preset gradient of the experimental design, the general nutrients of all raw materials were determined prior to formulation.
Table 1 test design
TABLE 2 NFC/NDF gradient diet formulas and nutrient levels
1) The premix provides for each kilogram of diet: vitamin a1,500IU; vitamin D550 IU; vitamin E10IU; iron 20mg; fierce, 40mg; zinc, 30mg; iodine 0.5mg; selenium 0.3mg; cobalt 0.2mg.
2) Besides the NFC/NDF value and the net energy of weight gain are calculated values, other nutrients are measured values.
1.4 preparation Process flow of fermented feed
1.4.1 production facility
A forage grass silk kneading machine, a movable transmission crawler belt, a full-mixing ration stirrer, a bacterial liquid spraying device, an automatic weighing device and a multifunctional fermentation barrel.
1.4.2 preparation Process flow
(1) Starting a transmission crawler belt, respectively conveying the whole broussonetia papyrifera and the corn silage with the water content of 70% into a pulverizer, pulverizing to 3cm, and conveying the crushed broussonetia papyrifera and the corn silage into a mixing stirrer through an automatic weighing device and the crawler belt;
(2) The refined materials with the water content of 13 percent are conveyed into a mixing stirrer through an automatic weighing device and a crawler belt;
(3) Taking a formula amount of the composite microbial inoculum in a microbial inoculum sprayer, and spraying the composite microbial inoculum into a mixing stirrer;
(4) Starting a mixing stirrer, stirring for 30min, and conveying to a pressing bucket through an inductor and a crawler belt for compaction treatment;
(5) And (3) conveying the compacted daily ration to a fermentation room through an automatic weighing device and a crawler belt, and fermenting for 30 days under the condition of room temperature and humidity kept at 45%, so as to obtain the paper mulberry type fermented total mixed daily ration.
1.5 sample collection and determination methods
Opening a vacuum bag at 30 days of fermentation, taking 200g of sample according to a 4-partition method, putting into a juicer, adding 1800mL of distilled water, stirring and mashing, filtering with 4 layers of gauze and qualitative filter paper to obtain a leaching solution, and immediately measuring the pH value of one part of the leaching solution and preserving the other part of the leaching solution at-20 ℃ for measuring organic acid; 500g of the sample is directly stored at-20 ℃ to measure the mycotoxin content.
FTMR feedstock profile nutrient: dry matter of raw materials (Dry matter, DM), crude Protein (CP), crude fat (Ether extract, EE), crude Ash (Crudeash, ash), neutral washing fiber (Neutral detergent fiber, NDF), acid washing fiber (Acid detergent fiber, ADF), calcium (Ca) and phosphorus (P) refer to Zhang Liying [16] The method of (2) is used for measurement.
And (3) organic acid content determination: the contents of Lactic Acid (LA), acetic Acid (AA), propionic Acid (PA) and Butyric Acid (BA) were determined by using an Shimadzu LC-20A type high performance liquid chromatograph, column chromatography: inertSustatin C18 (5 μm. Times.4.6X1250 nm); the mobile phase consisted of 0.05mol/L H pH 2.8 3 PO 4 -KH 2 PO 4 The buffer solution and acetonitrile are formed according to the volume ratio of 95:5, the flow rate is 0.5mL/min, the column temperature is 16 ℃, the wavelength of an ultraviolet detector is 210nm, and the sample injection volume is 10 mu L.
Determination of mycotoxin content: the content of Aflatoxin (AF), vomitoxin (DON) and Zearalenone (ZEA) is measured by enzyme-linked immunosorbent assay (ELISA) reagent, and the kit is purchased from Shanghai Kuailing Biotechnology Co., ltd, specific step reference Zhang Zhiguo and the like [17] Is reported in (3).
1.6 data statistics and analysis
The test data are collated by Excel, the test results are completely and randomly designed according to 2X 2 factors, the General Linear Model (GLM) program of SPSS18.0 software is called for variance analysis, and the main effects are the addition level of NFC/NDF and the complex bacteria and the interaction between the NFC/NDF and the complex bacteria respectively. The least squares mean of each treatment group was then subjected to multiple comparisons using the Duncan method, P <0.05 indicating significant differences. Finally, a 'Simple Correlation Analysis (SCA)' program is called to analyze the correlation between the organic acid and the mycotoxin content. The results are expressed as averages and the degree of variation in each treatment is expressed as Standard Error (SEM).
2 results and analysis
2.1 pH and organic acid content
Table 3 shows that, first, the substrate NFC/NDF values significantly affect the pH, lactic acid, acetic acid, propionic acid and butyric acid content of the fermentation substrate (P < 0.001), where the pH tends to decrease significantly linearly or secondarily with increasing NFC/NDF values (P < 0.05), with the highest pH value (P < 0.05) at NFC/ndf=1.08 and the lowest at NFC/ndf=1.60 (P < 0.05); the organic acid content was then linearly or secondarily increasing (P < 0.05), with the highest lactic and propionic acid content (P < 0.05) at NFC/ndf=1.60, the highest acetic acid at NFC/ndf=1.34, and the lower butyric acid (P < 0.05) at NFC/ndf=1.34 and 1.60.
Furthermore, the levels of complex bacteria added significantly affected the pH, lactic acid, propionic acid and butyric acid content of the fermentation substrate (P < 0.05), with the pH being highest in the non-added group (P < 0.05), the lactic acid content of the 3 complex bacteria added groups being higher (P < 0.05), the propionic acid content of the 4 and 8g/kg DM added groups being higher, and the butyric acid content being opposite (P < 0.05); the complex bacterial factors had no significant effect on the acetic acid content after substrate fermentation (p=0.271).
In addition, the diet structure has significant interactions with the complex bacteria additive factors (P < 0.05) on pH and organic acid content, with the pH of group A1 being highest, group C1 being lowest, lactic acid being higher in groups C3 and C4, group A1 being lowest, acetic acid being higher in groups C2 and C3, group A3 being lower, propionic acid being higher in group C3, group A1 being lower, and butyric acid being higher in groups A1 and A2, group C2 being lower.
TABLE 3pH and organic acid content of FTMR of each treatment group
Note that: the same column data shoulder marks have different letters representing significant differences (P < 0.05), and the same letters represent insignificant differences (P > 0.05)
2.2 mycotoxin content
Table 4 shows that the diet structure significantly affected the aflatoxin B1, vomitoxin and zearalenone content of the fermentation substrate (P < 0.001), the 3 mycotoxins content was all higher (P < 0.05) at NFC/ndf=1.08 for the substrate, the aflatoxin B1 was significantly linear and quadratic decreasing trend (P < 0.001), the substrate NFC/ndf=1.60 was lowest (P < 0.05); both vomitoxin and zearalenone content were in a secondary decrease trend (P < 0.001).
The complex microbial inoculant additive factors significantly affect the aflatoxin B1 and zearalenone content, wherein the aflatoxin B1 content is higher in the control group, and the zearalenone content of the 4 and 8g/kg DM complex microbial inoculant additive group is lower (P < 0.05).
Factor 2 has a significant interaction effect on aflatoxin B1 and zearalenone content (P < 0.05), with aflatoxin B1 highest in group A1 and D4, B4 and B2 lower (P < 0.05); similarly, the zearalenone content was highest in group A1, with lower groups B4 and B2 (P < 0.05).
Table 4 mycotoxin content of each treatment group
Note that: the same column data shoulder marks have different letters representing significant differences (P < 0.05), and the same letters represent insignificant differences (P > 0.05)
2.3pH and correlation of organic acids with mycotoxins
Table 5 shows that the pH of the fermentation substrate is very significantly positively correlated with the 3 mycotoxin levels (P < 0.01), whereas lactic acid is very significantly negatively correlated (P < 0.01), acetic acid and propionic acid are very significantly or significantly negatively correlated (P < 0.05) with aflatoxin B1, and butyric acid is very significantly positively correlated (P < 0.01) with aflatoxin B1 and zearalenone.
TABLE 5 correlation of organic acids with mycotoxins
Note that: * P <0.01, P <0.05.
Discussion 3
3.1pH and organic acid content
The pH value and the lactic acid content are important characterization indexes of the quality of the biological fermentation feed, wherein the reduction of the pH value is mainly caused by lactic acid accumulation, and when the pH value is lower than 4.20, the growth of harmful bacteria is strictly inhibited, thus achieving the purpose of stable storage [18] . The test result shows that under the condition of different carbohydrate structures (NFC/NDF values), the pH value of the fermentation substrate is lower than 4.20, and the substrate structure of each treatment group is basically reasonable; on the other hand, as the NFC/NDF value of the diet is increased, the pH value is in a trend of being secondarily reduced as a whole, and the lowest point appears in the NFC/NDF=1.60 group, which is probably caused by that the diet with high NFC content can provide a large amount of rapid fermentation substrates for lactobacillus under anaerobic condition to promote the lactobacillus to reproduce and grow and dominate, which is consistent with the results reported by most of the former people [19-21] However, this effect has a top effect, i.e. the highest NFC content does not achieve the lowest pH and highest lactic acid yield, probably because lactic acid bacteria preferentially utilize the high amounts of starch and soluble sugars in high amounts of NFC, rapidly promoting lactic acid accumulation, but at the same time prematurely feedback inhibiting the growth of lactic acid bacteria, so that lactic acid accumulation enters the stationary phase at a later and maximum. The NFC with a proper proportion can quickly utilize easily-fermented carbohydrate, the lactic acid yield does not reach the degree of strictly limiting the growth of the lactic acid bacteria, time is won for other slowly-released components in the NFC, accumulation of lactic acid is further promoted, and the mechanism needs to be further analyzed by dynamic research of the fermentation process. In addition, from the effect of adding the complex bacteria, the pH value of the test group is obviously lowIn the control group (no significant difference between the test groups), lactic acid production was in the opposite trend, again corroborating the results of the previous study: the exogenously added fermentation promoting microbial inoculum and lactobacillus contained in the material form a synergistic effect, thereby accelerating the fermentation process and promoting the fermentation degree [22] 。
Lactic acid, acetic acid, propionic acid and butyric acid are main metabolites produced by microorganisms in anaerobic fermentation feed by utilizing substrate nutrients (nitrogen source and carbon source), wherein lactic acid is the organic acid with the highest yield, the largest acidity and the largest molecular weight, and the higher the yield of lactic acid is, the deeper the fermentation degree of lactic acid bacteria is; the acetic acid and propionic acid yield are indicative of fermentation type, and the butyric acid yield is positively related to the activity level of spoilage bacteria, mold and butyric acid bacteria, i.e. the higher the butyric acid content is, the less the material is fermented, the larger the nutrition loss is, and the worse the quality is [23] . The test results show that the contents of lactic acid, acetic acid and propionic acid in each treatment have a tendency of a remarkable quadratic curve rising along with the rise of the NFC/NDF value, and are highest when NFC/NDF=1.60, and butyric acid has an opposite tendency, so that the carbon source structure of a substrate is one of the sensitive factors of the output of organic acid in an anaerobic fermentation mode, and the content of carbohydrate in rapid fermentation has a proper range. In addition, the acetic acid content of the composite bacterial agent treatment in the test is higher than that of the non-added group, the propionic acid content of the 4g/kg and 8g/kg added groups is higher, and the butyric acid content is lower, so that the composite bacterial agent has an additive dose effect, and the adding amount of 4g/kg is more suitable from the aspect of the adding cost.
3.2 mycotoxins
Mycotoxins are toxic secondary metabolites produced by certain fungal microorganisms under suitable conditions, and feed is subjected to fungal disease pollution during processing, storage and transportation, and the mycotoxins are produced, so that the quality of the feed is deteriorated; mycotoxins in forage grass products not only generate irreversible harm to livers, kidneys, immune systems and the like of livestock, but also remain in livestock products through accumulation of food chains, so that the safety risk of the livestock products is caused, and potential safety hazards are buried in the health of human bodies [24] . The fermented feed contains high moisture suitable for mold growth, so that detection of mycotoxins is necessary. The test focuses on the detection of aflatoxinB1, vomitoxin and zearalenone 3 are respectively 8.36-16.33ng/kg, 417-646ng/kg and 83.15-374.05ng/kg, which are lower than limit standard defined by national feed sanitation standard [25] (aflatoxin B1 is less than or equal to 20, vomitoxin is less than or equal to 3000 and zearalenone is less than or equal to 500 in other compound feeds), which shows that the anaerobic condition created by the test is more suitable. In addition, the carbohydrate structure of the fermentation substrate obviously influences the mycotoxin content in 3, the interaction of the addition amount of the composite microbial inoculum and the 2 factors has obvious influence on the aflatoxin B1 and zearalenone content, and the 2 test factors designed in the research show that the 2 test factors are important factors for regulating and controlling the mycotoxin production, and when NFC/NDF=1.60 of the fermentation substrate, the addition amount of the composite microbial inoculum is 4 and 8g/kg DM mycotoxin content is the lowest, so that the method has great guiding significance on actual production.
In addition, the correlation of mycotoxins, pH values and organic acid content of the broussonetia papyrifera fermented feed is analyzed, the pH value of a fermentation substrate is found to be extremely obviously positively correlated with 3 mycotoxin contents, lactic acid is extremely obviously negatively correlated, acetic acid and propionic acid are extremely obviously or obviously negatively correlated with aflatoxin B1, butyric acid is extremely obviously positively correlated with aflatoxin B1 and zearalenone, and the result accords with the theory reported by most of the prior people: the higher the organic acid content in the fermented feed in the anaerobic state, the stronger the ability of inhibiting the growth of harmful mold [26-27] Therefore, the organic acid content in the actual production can reflect the mould growth state of the fermented feed to a certain extent, and can be used as an important evaluation index of the fermentation quality.
Conclusion 4
1) The pH of the fermentation substrate showed a significant linear or quadratic decrease trend (P < 0.05) with increasing NFC/NDF, lowest at NFC/ndf=1.60 (P < 0.05); the organic acid content is linear or quadratic increasing (P < 0.05), wherein the lactic acid and propionic acid content is highest (P < 0.05) when NFC/NDF=1.60, the acetic acid is highest when NFC/NDF=1.34, and the butyric acid is lower (P < 0.05) when NFC/NDF=1.34 and 1.60; 2) The addition of the complex bacteria significantly reduces the pH value of the fermentation substrate (P < 0.05), improves the lactic acid content (P < 0.05), and has higher propionic acid content in 4 and 8g/kg DM addition groups, while the butyric acid content is opposite (P < 0.05); 3) The factor 2 has significant interactions on pH and organic acid content (P < 0.05), where pH is highest for group A1, lactic acid is higher for groups C3 and C4, acetic acid is higher for groups C2 and C3, propionic acid is higher for group C3, butyric acid is higher for groups A1 and A2; 4) The content of 3 mycotoxins is higher when the NFC/NDF of the substrate is=1.08 (P < 0.05), the aflatoxin B1 has a remarkable linear and secondary reduction trend (P < 0.001) along with the increase of the NFC/NDF, and the substrate is the lowest when the NFC/NDF of the substrate is=1.60 (P < 0.05); 5) The zearalenone content was lower in the 4 and 8g/kg DM complex plus group (P < 0.05), the 2 factor had a significant interaction effect on aflatoxin B1 and zearalenone content (P < 0.05), and was highest in the A1 group, lower in the B4 and B2 groups (P < 0.05); 6) The pH value of the fermentation substrate is in very significant positive correlation with the content of 3 mycotoxins (P < 0.01), while lactic acid is in very significant negative correlation (P < 0.01), acetic acid and propionic acid are in very significant or significant negative correlation with aflatoxin B1 (P < 0.05), and butyric acid is in very significant positive correlation with aflatoxin B1 and zearalenone content (P < 0.01). Therefore, the experiment finds that the NFC/NDF ratio, the addition of the composite microbial inoculum and the interaction of the two factors have obvious influence on the fermentation quality and the mycotoxin content of the complete mixed ration for the broussonetia papyrifera fermentation, the NFC/NDF proper ratio is 1.60, and the composite microbial inoculum proper addition amount is 4g/kg DM; the organic acid of the fermentation substrate is inversely related to the mycotoxin content.
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While the invention has been described in detail in the foregoing general description, with reference to specific embodiments and experiments, it will be apparent to one skilled in the art that modifications or improvements can be made thereto, and it is therefore intended that the invention as defined in the appended claims be construed as broadly as possible without departing from the spirit of the invention.
Claims (5)
1. The processing technology of the paper mulberry type fermented total mixed ration is characterized by comprising raw materials, concentrated materials and a composite microbial inoculum, wherein the raw materials comprise 30-50 parts of corn silage and 20-60 parts of whole paper mulberry; the fine material comprises 7-21 parts of corn, 1.7-4 parts of wheat bran, 0.9-5 parts of bean pulp, 0.5-1 part of corn dehydrated alcohol lees, 0.1-0.4 part of calcium carbonate, 0.08-0.3 part of calcium hydrophosphate, 0.04-0.2 part of sodium bicarbonate, 0.08-0.3 part of salt and 0.03-0.15 part of premix; the addition amount of the composite microbial inoculum is 3-5g/kg;
the processing technology of the paper mulberry type fermented total mixed ration comprises the following steps:
1) Starting a transmission crawler belt, respectively conveying the whole broussonetia papyrifera and the corn silage with the water content of 60-80% to a pulverizer, pulverizing to 2-3cm, and conveying to a mixing stirrer through an automatic weighing machine and the crawler belt;
2) The water content is divided into 12-15% concentrate, and the concentrate is conveyed into a mixing stirrer through an automatic weighing device and a crawler belt;
3) Taking a formula amount of the composite microbial inoculum in a microbial inoculum sprayer, and spraying the composite microbial inoculum into a mixing stirrer;
4) Starting a mixing stirrer, stirring for 20-40min, and conveying to a pressing bucket through an inductor and a crawler belt for compaction treatment;
5) Conveying the compacted daily ration to a fermentation room through an automatic weighing device and a crawler belt, fermenting for 20-40 days under the condition of room temperature and humidity kept at 40-50%, obtaining the paper mulberry type fermented total mixed daily ration,
the NFC/NDF value of the fermentation substrate of the paper mulberry type fermentation total mixed ration is 1.08-1.86, the composite microbial inoculum is lactobacillus plantarum and saccharomyces cerevisiae, and the addition amount is 2-10g/kg DM.
2. The processing technology of the paper mulberry type fermentation total mixed ration according to claim 1, wherein the paper mulberry type fermentation total mixed ration is composed of raw materials, concentrate and a composite microbial inoculum, and the raw materials comprise 42.47 parts of corn silage and 34.75 parts of whole paper mulberry; the concentrate comprises 15.50 parts of corn, 2.42 parts of wheat bran, 3.35 parts of soybean meal, 0.68 part of corn dehydrated alcohol lees, 0.27 part of calcium carbonate, 0.18 part of calcium hydrophosphate, 0.11 part of sodium bicarbonate, 0.18 part of salt and 0.08 part of premix; the addition amount of the composite microbial inoculum is 4g/kg.
3. The processing technology of the paper mulberry type fermented total mixed ration according to claim 2, wherein the processing technology of the paper mulberry type fermented total mixed ration is as follows:
1) Starting a transmission crawler belt, respectively conveying the whole broussonetia papyrifera and the corn silage with the water content of 70% into a pulverizer, pulverizing to 2.5cm, and conveying the whole broussonetia papyrifera and the corn silage into a mixing stirrer through an automatic weighing machine and the crawler belt;
2) The refined materials with the water content of 13 percent are conveyed into a mixing stirrer through an automatic weighing device and a crawler belt;
3) Taking a formula amount of the composite microbial inoculum in a microbial inoculum sprayer, and spraying the composite microbial inoculum into a mixing stirrer;
4) Starting a mixing stirrer, stirring for 30min, and conveying to a pressing bucket through an inductor and a crawler belt for compaction treatment;
5) And (3) conveying the compacted daily ration to a fermentation room through an automatic weighing device and a crawler belt, and fermenting for 30 days under the condition of room temperature and humidity kept at 45%, so as to obtain the paper mulberry type fermented total mixed daily ration.
4. The processing technology of the paper mulberry type fermented total mixed ration according to claim 1, wherein the NFC/NDF value of a fermentation substrate of the paper mulberry type fermented total mixed ration is 1.34-1.60, the composite microbial inoculum is lactobacillus plantarum and saccharomyces cerevisiae, and the addition amount is 4-8g/kg DM.
5. The processing technology of the paper mulberry type fermented total mixed ration according to claim 4, wherein the fermentation substrate NFC/NDF value of the paper mulberry type fermented total mixed ration is 1.60, the composite microbial inoculum is lactobacillus plantarum and saccharomyces cerevisiae, and the addition amount is 4g/kg DM.
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