CN116569989B - Biological organic selenium feed additive and preparation method and application thereof - Google Patents

Abstract

The invention discloses a biological organic selenium feed additive and a preparation method and application thereof. The invention comprises mixing peel residue, soybean meal, puffed soybean powder, wheat bran and sodium selenite, adjusting the moisture content to obtain the material and mixing it with plant lactobacillus, casei lactobacillus and animal bifidobacterium, fermenting, drying and crushing to obtain the biological organic selenium feed additive. The invention not only effectively recycles agricultural and forestry wastes such as peel residue, but also obtains a feed additive rich in biological organic selenium; moreover, the feed additive also contains rich probiotics, is beneficial to the growth of animals, and has good application prospects.

Description

A biological organic selenium feed additive and its preparation method and application

Technical Field

The invention belongs to the technical field of bioengineering, and particularly relates to a biological organic selenium feed additive and a preparation method and application thereof.

Background technique

With the continuous development of social productivity, human demand for meat protein has also increased. However, as the main source of meat protein, the productivity level of poultry and livestock farming is relatively low. Selenium (Se) is one of the trace elements necessary for animals to maintain their vitality. The lack of selenium can lead to slow growth of animals, nutritional metabolic diseases, and even estrus disorders, infertility, or weakness of newborn piglets after sow delivery. Appropriate supplementation of selenium in the diet can not only effectively avoid the occurrence of the above problems, but also improve the body's immune level and improve the reproductive performance of farmed animals, thereby improving the overall productivity of the poultry and livestock farming industry. According to the form of selenium, it can be divided into two forms: inorganic selenium and organic selenium. Inorganic selenium is more toxic, while the latter has higher biocompatibility and lower toxicity, and is the best form of selenium supplementation. At present, most feed production companies or farms add sodium selenite or yeast selenium to the feed. The former is more toxic and has a lower absorption rate. Large-scale use will have certain side effects. Although the use of yeast selenium can solve the problems of toxicity and absorption efficiency, the yeast selenium currently available on the market is expensive, which is a considerable expense for large-scale farms. Therefore, there is an urgent need to find a safe, widely available, inexpensive and easily absorbed animal selenium supplement.

Summary of the invention

The primary purpose of the present invention is to provide a method for preparing a biological organic selenium feed additive.

Another object of the present invention is to provide a biological organic selenium feed additive obtained by the above preparation method.

Another object of the present invention is to provide application of the above-mentioned biological organic selenium feed additive.

The purpose of the present invention is achieved by the following technical solution: A method for preparing a biological organic selenium feed additive comprises the following steps:

(1) mixing peel residue, soybean meal, puffed soybean powder, wheat bran and sodium selenite in a mass ratio of 500-800:100-300:50-100:10-30:0.5-2; adjusting the moisture content to 50%-70% with water to obtain material A;

(2) mixing the bacterial agent and the material A obtained in step (1) uniformly to obtain a fermentation system, and sealing and fermenting; the bacterial agent includes Lactobacillus plantarum, Lactobacillus casei and Bifidobacterium animalis;

(3) After the fermentation is completed, the mixture is dried and crushed to obtain a biological organic selenium feed additive.

The mass ratio described in step (1) is preferably 600-700:200-250:70-80:15-25:0.5-1.5; more preferably 650:220:75:20:1.

The peel residue described in step (1) is preferably citrus residue.

The water described in step (1) is preferably at least one of tap water and ultrafiltered tap water.

The moisture content in step (1) is preferably 60%.

In the bacterial agent described in step (2), Lactobacillus plantarum, Lactobacillus casei and Bifidobacterium animalis are preferably combined in a ratio of 1:1-3:1-3 in terms of cell number; more preferably, in a ratio of 1:1:1 in terms of cell number.

The bacterial agent described in step (2) is preferably obtained by combining an activated Lactobacillus plantarum bacterial solution, an activated Lactobacillus casei bacterial solution, and an activated Bifidobacterium animalis bacterial solution.

The concentration of the activated Lactobacillus plantarum liquid is preferably OD ₆₀₀ =1.0-1.4, and more preferably OD ₆₀₀ =1.2.

The concentration of the activated Lactobacillus casei bacterial solution is preferably OD ₆₀₀ =1.0-1.4, and more preferably OD ₆₀₀ =1.2.

The concentration of the activated animal Bifidobacterium bacterial solution is preferably OD ₆₀₀ =1.0-1.4; more preferably OD ₆₀₀ =1.2.

The bacterial agent is preferably prepared by the following steps: inoculating the preserved Lactobacillus plantarum, Lactobacillus casei and Bifidobacterium animalis into activation culture medium respectively, and then anaerobic culturing to obtain activated Lactobacillus plantarum, Lactobacillus casei and Bifidobacterium animalis bacterial liquid;

The activation medium consists of: peptone 10 g, beef extract 10 g, yeast extract 5 g, glucose 20 g, K ₂ HPO ₄ 2 g, NaOAc 5 g, MgSO ₄ 0.2 g, MnSO ₄ 0.05 g, Tween-80 1 mL, ammonium citrate tribasic 2 g, and distilled water to 1 L.

The inoculation amount of the inoculation is preferably 0.5-3% (v/v); more preferably 2% (v/v).

The temperature of the anaerobic culture is preferably 28-32°C; more preferably 30°C.

The addition amount of the bacterial agent described in step (2) is preferably calculated by adding 0.33-1.00mL of Lactobacillus plantarum liquid, 0.33-1.50mL of Lactobacillus casei liquid, and 0.33-1.50mL of Bifidobacterium animalis liquid in every 100g of material A; more preferably, 0.5mL of Lactobacillus plantarum liquid, 0.5mL of Lactobacillus casei liquid, and 0.5mL of Bifidobacterium animalis liquid are added in every 100g of material A.

The Lactobacillus plantarum is preferably Lactobacillus plantarum CCTCC WB 2008944.

The Lactobacillus casei is preferably Lactobacillus casei CCTCC KB 20081445.

The animal Bifidobacterium is preferably animal Bifidobacterium CGMCC 1.3003.

The sealed fermentation conditions in step (2) are preferably sealed fermentation at 33-39° C. for 5-9 days; more preferably sealed fermentation at 33-36° C. for 5-7 days.

The drying in step (3) is preferably carried out at a temperature below 50°C; more preferably at 45°C.

A biological organic selenium feed additive capable of regulating animal intestinal flora and improving animal immunity is obtained through the preparation method.

Application of the above biological organic selenium feed additive in preparing feed.

Compared with the prior art, the present invention has the following advantages and effects:

(1) The present invention provides a preparation process for obtaining an organic selenium feed additive by fermenting peel residue with composite probiotics, which can effectively recycle agricultural and forestry wastes such as peel residue, which contain a large amount of renewable resources. These renewable resources are not suitable for direct use by humans for various reasons. However, for microorganisms, these abundant agricultural and forestry wastes are excellent and cheap culture medium substitutes. Taking citrus residue as an example, these citrus residues discarded by the tangerine peel industry have relatively low sugar and acid content and poor palatability, and are not suitable for direct consumption. However, they contain a large amount of organic acids and sugar compounds mainly composed of citric acid, and microorganisms can directly and quickly use the nutrients therein for growth. In addition, citrus residue also contains a large amount of nutrients such as VC, which are also very beneficial to farmed animals.

(2) The three strains of probiotics used in the present invention are a combination carefully selected. First, plant lactobacillus and animal bifidobacterium are the main strains for converting organic selenium, but only combining plant lactobacillus and animal bifidobacterium can not obtain a particularly high conversion rate, and adding lactobacillus casei can further promote the bioconversion of organic selenium, and obtain an effect of 1+1+1 greater than 3. Secondly, lactobacillus casei and plant lactobacillus can produce a kind of biological preservative--lactobactin, which greatly improves the shelf life of this product, making this product easier to promote and apply. Finally, animal bifidobacterium has a very good effect of regulating intestinal flora, and has a good therapeutic effect on various acute and chronic diarrhea of poultry and livestock. In summary, the three strains of microorganisms in the present invention are synergistically fermented to obtain an excellent animal nutrition selenium supplement rich in organic selenium.

(3) The biological organic selenium feed additive in the present invention can provide safe and easily absorbed selenium elements for the animals being fed, and the probiotics contained therein can play a role in regulating the intestinal flora of the animals. The present invention can play an excellent role in the production process of various selenium-rich poultry and livestock products and has good application prospects. In addition, after the biological organic selenium feed additive provided by the present invention is absorbed by the animals, the waste discharged from the body also contains a relatively high level of selenium content, which can be used for the cultivation and production of selenium-rich agricultural and forestry crops, so as to form an efficient cycle development of selenium-rich agriculture and provide a positive effect on the construction of green ecology.

Detailed ways

The present invention is further described in detail below with reference to examples, but the embodiments of the present invention are not limited thereto.

Example 1

(1) Weigh the following raw materials in parts by weight: 650 parts of citrus pulp (the residue obtained after squeezing citrus juice), 220 parts of soybean meal, 75 parts of puffed soybean powder, 20 parts of wheat bran, and 1 part of sodium selenite.

(2) After the above raw materials are mixed, tap water is added until the moisture content is 60% to obtain material A.

(3) The activated Lactobacillus plantarum CCTCC WB 2008944 bacterial solution, Lactobacillus casei CCTCC KB20081445 bacterial solution and Bifidobacterium animalis CGMCC 1.3003 bacterial solution were mixed in a volume ratio of 1:1:1 to obtain a composite bacterial agent; then the composite bacterial agent was inoculated into material A at an inoculation amount of 1.5% (mL bacterial agent: g material A) and stirred again, and then sealed and fermented at 36° C. for 7 days.

The activation culture medium consists of: 10 g peptone, 10 g beef extract, 5 g yeast extract, 20 g glucose, 2 g K ₂ HPO ₄ , 5 g NaOAc, 0.2 g MgSO ₄ , 0.05 g MnSO ₄ , 1 mL Tween-80, 2 g triammonium citrate, and distilled water to 1 L.

The preserved Lactobacillus plantarum CCTCC WB2008944, Lactobacillus casei CCTCC KB20081445 and Bifidobacterium animalis CGMCC 1.3003 were inoculated into activation culture medium at a ratio of 2% (v/v), and then statically cultured anaerobically at 30°C until the _OD600 of each bacterial solution was 1.2 to obtain activated Lactobacillus plantarum CCTCC WB2008944 bacterial solution, Lactobacillus casei CCTCC KB20081445 bacterial solution and Bifidobacterium animalis CGMCC 1.3003 bacterial solution.

(4) Drying the fermented material at 45° C. and then crushing it to obtain a biological organic selenium feed additive.

Example 2

The process is basically the same as Example 1, except that the fermentation temperature is 33°C.

Example 3

The method is substantially the same as Example 1, except that the fermentation time is 5 days.

Example 4

The method is basically the same as Example 1, except that the fermentation temperature is 33° C. and the fermentation time is 5 days.

Comparative Example 1

The method is basically the same as Example 1, except that the bacterial solution used only contains Lactobacillus plantarum CCTCC WB2008944, wherein the inoculation amount of the bacterial solution of Lactobacillus plantarum CCTCC WB 2008944 is 1.5%, which is the same as the inoculation amount of the composite bacterial agent in Example 1.

Comparative Example 2

The method is substantially the same as that in Example 1, except that the bacterial solution used is only Lactobacillus casei CCTCC KB20081445, wherein the inoculation amount of the bacterial solution of Lactobacillus casei CCTCC KB 20081445 is 1.5%, which is the same as the inoculation amount of the composite bacterial agent in Example 1.

Comparative Example 3

The method is basically the same as Example 1, except that the bacterial solution used only contains Bifidobacterium animalis CGMCC1.3003, wherein the inoculation amount of Bifidobacterium animalis CGMCC 1.3003 bacterial solution is 1.5%, which is the same as the inoculation amount of the composite bacterial agent in Example 1.

Comparative Example 4

The invention is basically the same as the embodiment, except that the bacterial solution used only contains Lactobacillus plantarum CCTCC WB2008944 and Lactobacillus casei CCTCC KB 20081445, wherein the inoculum amounts of Lactobacillus plantarum CCTCC WB 2008944 and Lactobacillus casei CCTCC KB 20081445 are respectively 0.75%, and the sum of the inoculum amounts of the two is the same as the inoculum amount of the composite bacterial agent of embodiment 1.

Comparative Example 5

The method is basically the same as Example 1, except that corn kernels are used instead of citrus pulp.

Effect Example 1

The selenium content in the additives prepared in the examples and comparative examples was tested according to GB/T 13883-2008, and the results are shown in Table 1:

Table 1 Composition of additives in each embodiment

The ingredients of the additives prepared in each embodiment and comparative example are shown in Table 1. By comparing Example 1, Example 2, Example 3 and Example 4, when the culture time is 7 days, the organic selenium content in the feed additive fermented at 36°C is as high as 90.33% (1259.20 μg/g), while this value is only 66.75% (765.93 μg/g) when the fermentation temperature is 33°C; similarly, when the culture time is 5 days, the organic selenium content at 36°C fermentation temperature is 64.10% (765.93 μg/g), while at 33°C it is only 42.29% (434.02 μg/g). The above results show that temperature has a very important influence on the conversion of organic selenium. In addition, by comparing Example 1, Example 3 with Example 2, Example 4, when the fermentation temperature is controlled at 36°C, the organic selenium content after 7 days of culture is 90.33% (1259.20 μg/g), while that after 5 days of culture is only 64.10% (765.93 μg/g); similarly, when the culture temperature is 33°C, the organic selenium content in the additive after 7 days of fermentation is 66.75% (787.37 μg/g), while the organic selenium content after 5 days of fermentation is only 42.29% (434.02 μg/g). The above results indicate that as the fermentation time increases, organic selenium will gradually accumulate in the additive. In addition, by comparing Example 1 with Comparative Example 5, it was found that the organic selenium and protein contents obtained by fermenting corn kernels instead of citrus residues were at a relatively low level. This may be because citrus residues contain a large amount of organic acids and sugars, which can serve as a fast-acting carbon source for microbial growth, while the carbon source in corn kernels is mainly starch, which is relatively difficult for microorganisms to utilize. Therefore, after the same fermentation time, the microbial reproduction rate is not as good as that using citrus residues, and the converted organic selenium content is also relatively low. The relatively low protein content in feed additives under corn fermentation can also indicate that the bacterial content is relatively small.

On the other hand, by comparing Example 1 and Comparative Examples 1,2,3, it can be found that although the three probiotics (plant lactobacillus CCTCC WB 2008944, Lactobacillus casei CCTCC KB 20081445 and Bifidobacterium animalis CGMCC 1.3003) used in the present invention can all realize the synthesis of organic selenium under the fermentation condition of single strain, its synthesis efficiency is all generally not high, the highest plant lactobacillus CCTCC WB 2008944 also only has 71.20% (835.26 μ g/g) in the situation of single strain fermentation, and these three strains are carried out to synergistic fermentation under the same condition, its content of organic selenium can reach 90.33% (1259.20 μ g/g), show that these three probiotics may have certain synergy, when carrying out co-fermentation, can greatly improve the conversion efficiency of organic selenium.

Effect Example 2 Feeding and Test Results

180 healthy 21-week-old laying hens (Guangzhou Animal Husbandry Science Research Institute) were selected and randomly divided into three groups, Group A, Group B and Group C, with 60 in each group and 30 in a cage. Among them, Group A did not add biological organic selenium feed additives, Group B added 1% (w/w) of the biological organic selenium feed additive in Example 1 to the diet, and Group C added 5% of the biological organic selenium feed additive in Example 1 to the diet. The remaining conditions were carried out normally in accordance with the rules and regulations of the chicken farm. The test period was 5 weeks, with 1 week of pre-test and 4 weeks of formal test. The method for detecting selenium content in eggs and chicken is implemented in accordance with GB5009.93-2017.

The present invention adds 1% and 5% of the biological organic selenium feed additive in Example 1 to the diet of group B and group C respectively through comparative tests. The selenium content in the three groups of eggs was detected respectively, and it was found that the average selenium content in the eggs produced in group A was 0.115 μg/g, while the average selenium content in the eggs of group B and group C was 1.72 and 2.22 μg/g, respectively, which was significantly higher than the average selenium content in the eggs of group A (p < 0.05). In addition, the organic selenium feed additive had no significant effect on the egg production rate, average egg weight and survival rate of laying hens (p > 0.05), but the laying hens in group B were able to eat all the feed in advance before the next feeding, while the laying hens in group C were just the opposite. The laying hens in group C had a part of the feed left before the next feeding, which showed that a small amount of this product may have the effect of promoting the feeding of laying hens. By detecting the selenium content of the main parts of the whole body of the tested laying hens, it was found that the organic selenium feed additive can significantly increase the selenium content of various parts of the laying hens (p < 0.05), forming selenium-rich chicken. On the other hand, the feces of these selenium-enriched laying hens have a high selenium content, which can be further made into selenium-enriched organic fertilizer for cultivating selenium-enriched agricultural and forestry crops, thus achieving efficient circular development in the field of selenium-enriched. The above results show that the biological organic selenium feed additive provided by the present invention has a wide range of application prospects.

Table 2 Effects of adding different proportions of organic selenium feed additives to the diet on the selenium content in eggs

Note: “*” indicates significant difference between the groups, p < 0.05.

Table 3 Effects of adding different proportions of organic selenium feed additives to the diet on the production performance of laying hens

Note: "0" means that the test laying hens have eaten all the feed at normal time, "+" means that the test laying hens have eaten all the feed before the next feeding, and "-" means that the test laying hens have not eaten all the feed at the next feeding.

Table 4 Effects of adding different proportions of organic selenium feed additives to the diet on the selenium content in laying hens

The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims (8)

1. A method for preparing a biological organic selenium feed additive, characterized in that it comprises the following steps: (1) Mix peel residue, soybean meal, puffed soybean powder, wheat bran and sodium selenite in a mass ratio of 500-800:100-300:50-100:10-30:0.5-2; adjust the moisture content to 50%-70% with water to obtain material A; (2) mixing the bacterial agent and the material A obtained in step (1) uniformly to obtain a fermentation system, and sealing the system for fermentation; the bacterial agent includes Lactobacillus plantarum, Lactobacillus casei and Bifidobacterium animalis; (3) drying and crushing after fermentation to obtain a biological organic selenium feed additive; The bacterial agent in step (2) is compounded in a ratio of 1:1-3:1-3 in terms of cell number; The plant lactobacillus is plant lactobacillus CCTCC WB 2008944; The Lactobacillus casei is Lactobacillus casei CCTCC KB 20081445; The animal bifidobacterium is animal bifidobacterium CGMCC 1.3003; The sealed fermentation conditions in step (2) are sealed fermentation at 36° C. for 7 to 9 days. 2. The method for preparing the biological organic selenium feed additive according to claim 1, characterized in that: The mass ratio described in step (1) is 600-700:200-250:70-80:15-25:0.5-1.5. 3. The method for preparing the biological organic selenium feed additive according to claim 1, characterized in that: The peel residue in step (1) is citrus residue; The water described in step (1) is at least one of tap water and ultrafiltered tap water. 4. The method for preparing the biological organic selenium feed additive according to claim 1, characterized in that: The bacterial agent in step (2) is obtained by combining an activated Lactobacillus plantarum bacterial solution, an activated Lactobacillus casei bacterial solution, and an activated Bifidobacterium animalis bacterial solution; The concentration of the activated Lactobacillus plantarum bacterial solution is OD ₆₀₀ = 1.0-1.4; The concentration of the activated Lactobacillus casei bacterial solution is OD ₆₀₀ = 1.0-1.4; The concentration of the animal Bifidobacterium bacterial solution obtained by the activation is OD ₆₀₀ = 1.0-1.4; The amount of the bacterial agent added in step (2) is calculated by adding 0.33-1.00 mL of Lactobacillus plantarum solution, 0.33-1.50 mL of Lactobacillus casei solution, and 0.33-1.50 mL of Bifidobacterium animalis solution to every 100 g of material A. 5. The method for preparing the biological organic selenium feed additive according to claim 4, characterized in that: The bacterial agent is prepared by the following steps: inoculating the preserved plant lactobacillus, casei lactobacillus and animal bifidobacterium into activation culture medium respectively, and then anaerobic culture is performed to obtain activated plant lactobacillus liquid, casei lactobacillus liquid and animal bifidobacterium liquid; The activation medium consisted of: peptone 10 g, beef extract 10 g, yeast extract 5 g, glucose 20 g, K ₂ HPO ₄ 2 g, NaOAc 5 g, MgSO ₄ 0.2 g, MnSO ₄ 0.05 g, Tween-80 1 mL, ammonium citrate tribasic 2 g, and distilled water to 1 L. 6. The method for preparing the biological organic selenium feed additive according to claim 1, characterized in that: The drying in step (3) is drying at a temperature below 50°C. 7. A biological organic selenium feed additive, characterized in that it is obtained by the preparation method described in any one of claims 1 to 6. 8. Use of the biological organic selenium feed additive according to claim 7 in preparing feed.