CN112806478A - Production and application effect evaluation of microbial protein feed mainly containing tea seed meal - Google Patents
Production and application effect evaluation of microbial protein feed mainly containing tea seed meal Download PDFInfo
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Abstract
The invention discloses a production method and nutritional value evaluation of a microbial protein feed mainly containing tea seed meal. In order to reduce the crude fiber of the tea seed meal and improve the quality of crude protein of the tea seed meal, the feed is prepared by taking the tea seed meal as a substrate and performing semi-solid fermentation on aspergillus niger. According to a single-factor test scheme, the optimal proportion of fermentation conditions such as fermentation temperature (18-37 ℃), fermentation time (1-5d), moisture content (5-25%), charging amount (20-100g), inoculation proportion (5-25%) and the like is optimized. The fermentation temperature, the charging amount and the fermentation time influence the crude protein and crude fat content of the fermented tea seed meal; the inoculation ratio has obvious influence on the crude fiber and the total energy of the fermented tea seed meal; the fermentation time and the fermentation temperature affect the calcium content of the fermented tea seed meal. Under the optimal condition, the neutral detergent fiber content of the tea seed meal is reduced to 40.63%, and the content of crude fat, crude protein and calcium is increased. The apparent total intestinal metabolic rate of the conventional nutrients and amino acids of the fermented tea seed meal in the chicken is not different from that of the unfermented group.
Description
Technical Field
The invention relates to the field of microbial protein feed production by taking tea seed meal as a substrate, in particular to biological feed production for improving the nutritional value of the tea seed meal through microbial fermentation, and belongs to the technical field of feeds.
Background
China is the first major pork producing and consuming countries in the world, and the pork yield and consumption account for more than half of the total amount of the world. However, while the pig industry is rapidly developing, the problems of feed resource shortage and food competition of the pigs and the people are also met. More than 50% of the total domestic food production and its processing by-products have been used as feed. Moreover, in recent years, the food import of China is in the trend of rising year by year: in 2018, the corn, the wheat, the barley, the sorghum and the DDGS are imported with 1723.6 ten thousand tons in total, the imported soybean is 8803 ten thousand tons, the soybean meal is 2.27 ten thousand tons, the rapeseed meal is 130.2 ten thousand tons, and the fish meal and the whey powder are imported with 146.1 ten thousand tons and 55.5 ten thousand tons respectively. In imported soybeans and grains, more than 70% of the soybeans and grains are used as feed, and it is seen that the international dependence of China on feed raw materials reaches a quite high level.
The culture area of the camellia oleifera in China exceeds 347 ten thousand hectares, the annual tea seed yield can reach 100 ten thousand tons, and the annual tea seed meal can reach 55 ten thousand tons calculated by the extraction meal yield of the tea seed cake of 94 percent. The tea seed cake contains crude protein 10.89-15.78%, crude fat 1.25-6.82%, saccharide 33.6-36.49%, and crude fiber 6.9-39.08%, wherein the content of tea saponin is 5.63-24.06%, the water content is 8.22-9.45%, and the ash content is 2.97-5.53%. Besides, it is rich in compounds such as protein, polyphenol, polysaccharide and saponin. The tea seed meal is mainly rich in 17 amino acids and contains various trace elements such as Fe, Cu, Zn, K, Mn, Mg, Ca and the like. The plant cell wall mainly comprises crude fibers, wherein the components such as cellulose, hemicellulose, lignin, cutin and the like contained in the crude fibers can prevent digestive enzymes from directly contacting with chyme, so that the nutrient digestibility is reduced, and in addition, the crude fibers can also prevent small molecules from being absorbed in intestinal tracts. Therefore, the application of the tea seed meal in the livestock and poultry production is limited, and a method for effectively reducing the content of crude fiber and tea saponin in the tea seed meal is urgently needed, so that the nutritive value of the tea seed meal in the livestock and poultry is improved.
The microbial protein feed can improve the conversion utilization rate of the feed. The beneficial microorganisms can enhance the digestive ability of digestive tract microorganisms and promote the digestion, absorption and utilization of nutrients by animals. And most of the fermented feed has sauce flavor and good palatability, and the feed intake of the livestock and poultry is increased. In addition, the microorganisms produce acetic acid, propionic acid, lactic acid and other organic acids during fermentation, and these acids can inhibit the growth of harmful microorganisms such as colibacillus and other harmful microorganisms in vivo and in vitro, and raise the disease resistance of animal. After the camellia seed meal is subjected to solid-state fermentation by using bacillus subtilis, candida utilis and aspergillus niger, the degradation rates of tea saponin, crude fiber and total phenol in the camellia seed meal respectively reach 87.63%, 37.29% and 87.87%, and the improvement rate of crude protein even reaches 66.98%. Aspergillus niger, Candida utilis, mucor and the like are used for carrying out solid state fermentation on the tea seed meal, and the strain can not only grow normally on the detoxified tea seed meal, but also can improve the protein content of a fermentation product.
Based on the research progress, the tea seed meal is taken as a main raw material, and through systematic research on the influence of different fermentation conditions on the nutritional value of the tea seed meal, a proper strain is searched, the fiber content of the tea seed meal is reduced, the protein quality of the tea seed meal is improved, and a reasonable mode for producing the tea seed meal fermentation microbial protein feed through fermentation is established. Further evaluating the nutritional value of the fermented tea seed meal microbial protein feed on poultry, and providing a more practical theoretical and practical basis for the production of feed in China. The method has profound significance for relieving the problems of feed resource shortage, environmental pollution caused by tea seed meal and the like.
Disclosure of Invention
The invention aims to provide a production method of a microbial protein feed and an application effect thereof, which are used for solving the problems in the prior art and improving the nutritive value of an industrial byproduct, namely tea seed meal.
In order to achieve the purpose, the invention provides the following scheme: the invention provides a production method of microbial protein feed, wherein Aspergillus niger is used as a fermentation strain, tea seed meal is used as a fermentation substrate, and the proper conditions of the fermentation conditions are respectively as follows:
the fermentation temperature (18-37 ℃) is 4.27-8.05 percent of crude protein of the fermented tea seed meal, 12.83-16.38 percent of crude fat, 0.19-0.37 percent of calcium and 0.135-0.1425 percent of phosphorus;
fermenting for 1-5 days, wherein the fermented tea seed meal crude protein is 6.51-17.90%, the crude fat is 12.84-17.38%, the calcium is 0.24-0.89%, and the phosphorus is 0.12-0.16%;
the water content (5-25%), the crude protein of the fermented tea seed meal is 7.28-8.19%, the crude fat is 12.02-14.23%, the crude fiber is 57.63-60.11%, and the total weight is 18.41-19.75 KJ/g;
the charging amount (20-100g), at this time, the fermented tea seed meal crude protein is 5.36-8.60%, the crude fat is 8.90-11.50%, the crude fiber is 58.14-63.90%, and the total amount can be 20.2-21.06 KJ/g;
the inoculation proportion (5-25%), 8.62-10.29% of crude protein of the fermented tea seed meal, 6.45-9.90% of crude fat and 62.03-65.22% of crude fiber, and the total amount can be 19.43-20.36 KJ/g;
the microbial protein feed disclosed by the invention has the effects of improving the content of crude protein and crude fat, reducing the content of neutral detergent fiber and increasing the content of calcium, and metabolic tests on Xianghuang chickens show that the apparent metabolic rates of nutrient components of fermented tea seed meal and unfermented tea seed meal are not obviously different.
The invention discloses the following technical effects: compared with tea seed meal, the fermented tea seed meal has the advantages that the crude protein, crude fat and calcium are obviously increased, the neutral detergent fiber content is reduced, and the tea saponin has no obvious difference. After fermentation, the total energy, dry matter, crude fat and organic matter total intestinal apparent metabolic rate of the fermented tea seed meal is not influenced (P is less than 0.05).
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the attached tables in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, the present invention is described in detail with reference to the accompanying tables and embodiments.
Example 1
1 materials and methods
1.1 activation of the bacteria and preparation of the fermentation Medium
Aspergillus niger is provided by the institute of Life sciences and Environment of the Yang university academy, and is activated on a potato agar slant culture medium at 28 ℃ for 7 days, then a single colony is inoculated into a potato liquid seed culture medium, and is subjected to shake cultivation at 28 ℃ and 180r/min for 2 d. And filtering the Aspergillus niger spore liquid which is cultured for 2d and is in the logarithmic growth phase by eight layers of sterile gauze to obtain spore suspension. Tea seed meal is a byproduct of tea oil produced by tea seeds by a manual squeezing method and is given away by oil extraction workshops. Pulverizing tea seed cake, sieving with 40 mesh sieve, adding appropriate amount of water, and treating at 0.2MP 120 deg.C for 20 min.
1.2 fermentation Condition optimization
Adding tea seed meal into a 250mL triangular flask, adding water, sterilizing at 120 ℃ for 20min, adding bacterial liquid, culturing at 28 ℃ for several days, and collecting a product to be tested. In the fermentation process, single-factor tests are carried out on the fermentation temperature (18-37 ℃), the fermentation time (1-5d), the moisture content (5-25%), the loading amount (20-100g) and the inoculation ratio (5-25%), wherein each factor is set to be 5 levels, and each level is set to be 6 times. And (3) obtaining the optimal fermentation condition by taking the reduction amplitude of the crude fiber and the improvement amplitude of the crude protein as optimization targets.
1.3 sample Collection analysis
Drying the fermented product at 65 deg.C to constant weight, sieving with 40 mesh sieve, and storing in refrigerator at-20 deg.C. The crude protein is detected by referring to a Kjeldahl method, crude fat is measured by referring to GB/T6433-[8]. The measurement of crude ash content, hydrochloric acid insoluble ash content, calcium and phosphorus can be performed according to feed analysis and feed quality control written by ZhangieyingDetection technology detection.
1.4 statistical analysis of data
The results were analyzed using the GLM program of SAS8.0 software. When the difference is significant, the mean between groups is subjected to multiple comparisons by the Duncan method, and the result is expressed as mean value. + -. standard error (or standard deviation). Differences were considered significant when P < 0.05.
2 results and analysis
The results show that the fermentation temperature significantly affects the content of crude protein, crude fat and calcium of the fermented tea seed meal (P <0.05), and has no significant effect on the content of phosphorus (P > 0.05). The water content ratio has no significant influence on the crude protein, crude fat, crude fiber and total energy of the fermented tea seed meal (P > 0.05). The loading amount significantly affected the crude protein and crude fat content of the fermented tea seed meal (P <0.05), and had no significant effect on the crude fiber and total energy content (P > 0.05). The inoculation ratio has a significant effect on the crude fiber and total energy of the fermented tea seed meal (P <0.05) and has no significant effect on the crude protein and crude fat content (P > 0.05). The fermentation time obviously influences the crude protein and fat content, calcium content and phosphorus content (P <0.05) of the fermented tea seed meal.
After fermentation, the crude fat of the tea seed meal is remarkably increased, the crude protein is increased to 7.88 +/-0.35% from 6.03 +/-1.21% (P <0.05), the total content is increased to 20.56 +/-0.43% (P <0.05) from 19.70 +/-0.71%, the calcium content is increased to 0.58 +/-0.27% (P <0.05) from 0.23 +/-0.04%, the neutral detergent fiber is reduced to 19.24% from 32.41, and the amplitude is reduced to 40.63%.
Example 2
Test animal selection and rearing management
48 Xiang yellow chickens of the same age of the day and the same variety of 30 days are selected as test animals, and the test chicken house is positioned in an animal house of the Yang-balancing teachers and universities, and is naturally ventilated and sufficiently illuminated. In the experiment, Xiang yellow chickens were raised in a three-dimensional single cage, and were fed once in the morning and evening every day without any restriction on drinking water. Sanitary cleaning and disinfection are carried out on time every day. Feces and urine were collected daily and the health of the flock was observed.
2 design of the experiment
2.1 daily ration and animals
The experiment was divided into control and experimental groups, each group was 12 replicates, with 4 chickens per replicate. The group A was fed with basal diet (see Table 1), the group B was fed with 20% of tea seed meal, and the group C was fed with 20% of fermented tea seed meal. The nutrient metabolism rate test was performed by the alternative method.
2.2 sample Collection and processing
After 3 days of feed transition period, continuously collecting feces and urine for 4 days, and dripping sulfuric acid in time after each feces collection to prevent ammonia and nitrogen from volatilizing; and adding toluene dropwise for treatment for corrosion prevention. Mixing the collected excrement and urine every day, drying at 65 ℃, fully grinding by using a mortar, sieving by using a 40-mesh sieve, and placing into a refrigerator at-20 ℃ for storage to be tested.
2.3 index detection
The total energy, crude protein, crude fiber, calcium, phosphorus and crude ash content in the tea seed meal, the fermented tea seed meal, the feed and the excrement are detected by referring to the book of feed analysis and feed quality detection technology. The hydrolyzed amino acids were determined using an amino acid analyzer.
And (3) calculating the nutrient apparent metabolic rate of the fermented tea seed meal according to a substitution method according to books of animal nutriology of Mr. Yang Feng.
2.4 data processing and analysis
The experimental data were single factor anova using SAS8.0 software, the differences between groups were tested using Duncan, and the results were expressed as mean ± standard error (mean ± SE), with a significance level P < 0.05.
3 results and analysis of the experiments
Compared with the tea seed meal, the total energy, crude protein, crude fat, crude ash and calcium content of the fermented tea seed meal are obviously increased (P is less than 0.05), the neutral detergent fiber is reduced from 32.41 to 19.24 percent, the reduction amplitude reaches 40.63 percent, and the content of tea saponin is reduced (Table 2). After fermentation, the total energy, dry matter, crude fat, total intestinal apparent metabolic rate of organic matter of the tea seed meal was unaffected (P >0.05) (table 3).
The production method of the biological feed has the effects of reducing the content of neutral washing fiber of the tea seed meal and increasing the content of crude protein, crude fat and phosphorus of the tea seed meal.
The above is only one embodiment of the present invention, and it should be noted that, for those skilled in the art, several similar modifications and improvements can be made without departing from the inventive concept of the present invention, and these should also be considered as within the protection scope of the present invention.
TABLE 1 broiler basic diet composition and nutritional level (air-dry basis)
Table 1 Composition and nutrient levels of basal diets(air-dry basis)
Note: premix feed to each kilogram of diet: VA 11000 IU; VC is 15 mg; VD 32000 IU; VE 1 mg; VK 31 mg; VB 11 mg; VB 25 mg; VB60.8mg; VB120.001mg; folic acid 0.2 mg; nicotinic acid 2.5 mg; biotin 0.5 μ g; 8mg of copper; 60mg of manganese; 80mg of iron; 40mg of zinc; 0.35mg of iodine; selenium 0.15mg.
TABLE 2 conventional nutrient content and amino acid composition of tea seed meal and fermented tea seed meal
Table 2.Nutrients concentration and amino acid of camellia oleifera seed meal(COM)and fermented COM
Remarking: the detection of neutral detergent fiber, acidic detergent fiber and hydrolyzed amino acid is carried out by the subtropical agroecological research institute of Chinese academy of sciences, and is the result of detection after mixing six samples
TABLE 3 Total intestinal apparent metabolic rate of tea seed meal and fermented tea seed meal
Table 3 In vivo nutrients total intestinal apparent digestibility of camellia oleifera seed meal(COM)and fermented COM
Claims (3)
1. A fermentation culture medium of microbial protein feed mainly containing tea seed meal is characterized in that the tea seed meal is mainly added with water and then treated for 20min at 120 ℃. The fermentation medium comprises the following components: 75-95% of tea seed meal and 5-25% of water.
2. A process for preparing a microbial protein feed according to claim 1, comprising the steps of:
(1) activating Aspergillus niger on potato solid culture medium for 4 days until mycelia grow vigorously, collecting flat plate, cleaning with sterilized distilled water, and filtering with eight layers of gauze to obtain spore.
(2) Adding 5-25% water into fermentation culture medium, treating at 120 deg.C for 20min in high temperature high pressure sterilizing pot, inoculating 5-25% mixed bacterial spore suspension, and fermenting at 18-37 deg.C for 1-5d to obtain biological feed.
(3) The result shows that the fermentation temperature obviously influences the content of crude protein, crude fat and calcium of the fermented tea seed meal (P <0.05), and has no obvious influence on the content of phosphorus (P > 0.05); the water content ratio has no obvious influence on the crude protein, crude fat and crude fiber of the fermented tea seed meal (P is more than 0.05); the charging amount significantly influences the crude protein and crude fat content of the fermented tea seed meal (P <0.05), and has no significant influence on crude fiber and total energy content (P > 0.05); the inoculation ratio has obvious influence on the crude fiber and the total energy of the fermented tea seed meal (P <0.05), and has no obvious influence on the content of crude protein and crude fat (P > 0.05); the fermentation time obviously influences the crude protein and fat content, calcium content and phosphorus content (P <0.05) of the fermented tea seed meal.
3. The nutritional value of the biological feed made according to claims 1 and 2, characterized in that:
(1) compared with the unfermented feed, after the fermentation of the fermented feed, the crude fat of the tea seed meal is remarkably increased, the crude protein is increased to 7.88 +/-0.35% from 6.03 +/-1.21% (P <0.05), the total content is increased to 20.56 +/-0.43% (P <0.05) from 19.70 +/-0.71%, the calcium content is increased to 0.58 +/-0.27% (P <0.05), the neutral detergent fiber is reduced to 19.24% from 32.41, and the reduction amplitude is 40.63%.
(2) The nutrient total intestinal tract apparent metabolic rate of the fermented tea seed meal is evaluated on broiler chickens by a substitution method, and compared with unfermented tea seed meal, the metabolic rates of various conventional nutrients and amino acids of the fermented tea seed meal have no significant difference.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103039711A (en) * | 2013-01-20 | 2013-04-17 | 湖南奇异生物科技有限公司 | Preparation method of camellia seed dreg fermented feed |
| CN103211084A (en) * | 2013-04-17 | 2013-07-24 | 安徽农业大学 | Method for preparing biological enzyme-enriched camellia seed meal fermented feed |
| CN110897032A (en) * | 2019-11-19 | 2020-03-24 | 华南理工大学 | Fermented feed protein and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103039711A (en) * | 2013-01-20 | 2013-04-17 | 湖南奇异生物科技有限公司 | Preparation method of camellia seed dreg fermented feed |
| CN103211084A (en) * | 2013-04-17 | 2013-07-24 | 安徽农业大学 | Method for preparing biological enzyme-enriched camellia seed meal fermented feed |
| CN110897032A (en) * | 2019-11-19 | 2020-03-24 | 华南理工大学 | Fermented feed protein and preparation method and application thereof |
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