CN102919516A - Novel vinasse biological feed and production process thereof - Google Patents

Abstract

The invention discloses a new type of distiller's grains biological feed and its production process, including distiller's grains medium and strains, characterized in that the formula of distiller's grains medium is: 50%-70% of fresh distiller's grains, 5%-15% of corn flour , rapeseed meal 5% ~ 15%, water 15% ~ 25%, the percentage here refers to the ratio of each formula to the total weight of distiller's grain medium; add ferrous sulfate 12 ~ 18mg to every 80 ~ 120g distiller's grain medium , zinc sulfate 10-15mg, manganese sulfate 8-12mg, copper sulfate 12-18mg, calcium hydrogen phosphate 1-3g; strains include Bacillus subtilis, Trichoderma viride, Lactobacillus acidophilus and Brettanomyces bantu; process flow Including: Antagonism test, extended culture of bacteria, purity and activity inspection, preparation and sterilization of distiller's grain medium, solid-state fermentation of distiller's grain microorganisms and treatment of distiller's grain biological feed after 9-11 days; short cycle and high protein content.

Description

A new type of distiller's grain biological feed and its production process

technical field

The invention relates to a production process for processing distiller's grains to produce novel biological feedstuffs from distiller's grains.

Background technique

The annual output of distiller's grains in my country is as high as more than 21 million tons. Based on the fact that distiller's grains contain certain protein, rich in amino acids, vitamins and various trace elements, etc., the traditional method is to feed fresh distiller's grains directly to livestock as feed raw materials. There are two methods for this method: There are two disadvantages: first, the crude fiber content in fresh distiller's grains is high, the palatability of livestock is poor, the digestibility is low, and moldy distiller's grains are prone to pests and diseases; second, fresh distiller's grains have high moisture content, are difficult to transport, have high acidity, and are easy to spoil. For this reason, people have carried out various studies on its resource utilization, such as: using distiller's grains to produce glycerin, cultivating edible fungi, extracting compound amino acids and trace elements, extracting phytic acid and calcium phytate, and brewing vinegar; using distiller's grains Anaerobic fermentation recovers biogas, extracts protein, produces dry whole feed (DDGS), produces single-cell protein (SCP) feed, distillers grains ferments to produce fuel ethanol, and produces crude enzyme preparations, etc. Most of the above methods are at a standstill due to problems such as small scale, large energy consumption, and insufficient conversion of nutrients.

Contents of the invention

The technical problem to be solved by the present invention is to provide a production process for effectively fermenting distiller's grains to produce distiller's grains biological feed, so as to overcome the shortcomings of the prior art such as large energy consumption and insufficient conversion of nutrients.

The technical scheme of the present invention: a new type of distiller's grains biological feed, including distiller's grains medium and strains, characterized in that the formula of distiller's grains medium is: 50%-70% of fresh distiller's grains, 5%-15% of corn flour, vegetable 5% to 15% of seed meal, 15% to 25% of water, the percentage here refers to the ratio of each formula to the total weight of distiller's grain medium; and every 80 to 120 g of distiller's grain medium is added with 12 to 18% ferrous sulfate mg, zinc sulfate 10-15 mg, manganese sulfate 8-12 mg, copper sulfate 12-18 mg, calcium hydrogen phosphate 1-3g; strains include Bacillus subtilis, Trichoderma viride, Lactobacillus acidophilus and Bantu yeast.

For every 80-120 g of distiller's grains medium, 1-3 mL of Bacillus subtilis bacterial liquid is inoculated, and the number of viable bacteria is 1.0×10 ¹¹ to 1.5×10 ¹¹ cfu/mL.

Every 80-120 g of distiller's grains medium is inoculated with 1-3 mL of Trichoderma viride bacterial liquid, and the number of viable bacteria is 6.8×10 ¹⁰ to 7.2×10 ¹⁰ cfu/mL.

For every 80-120 g of distiller's grains medium, 1-3 mL of Lactobacillus acidophilus liquid is inoculated, and the number of live bacteria is 2.0×10 ¹⁰ to 2.2×10 ¹⁰ cfu/mL.

For every 80-120 g of distiller's grains medium, 1-3 mL of Bantu Brettanomyces yeast liquid is inoculated, and the number of viable bacteria is 3.2×10 ¹¹ to 3.6×10 ¹¹ cfu/mL.

The production process of a new type of distiller's grain biological feed as described above is characterized in that it includes the following processes: conducting antagonism test, strain expansion culture, purity and activity inspection, preparation and sterilization of distiller's grain medium, solid-state fermentation of distiller's grain microorganisms and 9 ~11 d post-distiller's grain bio-feed treatment.

The above-mentioned antagonism test: divide the 4 species of Brettanomyces bantu, Bacillus subtilis, Trichoderma viride, and Lactobacillus acidophilus into groups in pairs, and draw one of the bacteria in each group at an interval of 8-12 mm Inoculate it on ordinary medium, then take another kind of bacteria and draw a straight line vertically, incubate at 28-32°C for 22-26 hours, and observe whether the intersection is inhibited and the colony shrinks and disappears.

Expansion culture, purity and activity inspection of the described strains: Inoculate a ring of slant bacteria into 100mL liquid culture medium with 4 selected strains respectively, shake the strains at 150r/min and 30°C, in which Bacillus subtilis and Lactobacillus acidophilus After 10-14 hours of expanded cultivation, they are transferred to fermenters for further expanded cultivation. After 22-26 hours of expanded cultivation, Trichoderma viride and Bantu Brettanomyces are transferred to fermentors for further expanded cultivation. During the expanded culture process, samples were taken from the fermenter for Gram staining and observed under a microscope to determine whether it was the target strain. At the same time, the colony morphology and colony count were observed by coating to determine the activity of the strain.

The preparation and sterilization of the distiller's grain medium: 50% to 70% of fresh distiller's grains, 5% to 15% of corn flour, 5% to 15% of rapeseed meal, and 15% to 25% of water, the percentages here Refers to the ratio of each formula to the total weight of the distiller's grain medium; and for every 80-120 g of distiller's grain medium, add 12-18 mg of ferrous sulfate, 10-15 mg of zinc sulfate, 8-12 mg of manganese sulfate, and 12 mg of copper sulfate ～18 mg, 1～3g of calcium hydrogen phosphate, add lime water to adjust the pH to 6～7, mix well, and stack at normal pressure for sterilization.

The solid-state fermentation of distiller's grain microorganisms: when the temperature of the distiller's grain medium drops to 32-38°C, inoculate the 4 kinds of bacteria expanded and cultivated in the fermenter into the distiller's grain medium in equal proportions, with a total inoculation amount of 5%-7%. middle. The viable count of Bacillus subtilis is 1.0×10 ¹¹ ~1.5×10 ¹¹ cfu/mL, the viable count of Trichoderma viride is 6.8×10 ¹⁰ ~7.2×10 ¹⁰ cfu/mL, and the viable count of Lactobacillus acidophilus is 2.0× 10 ¹⁰ ～2.2×10 ¹⁰ cfu/mL, and the viable count of Brettanomyces bantu was 3.2×10 ¹¹ ～3.6×10 ¹¹ cfu/mL. After inoculation, when the temperature drops to 28-32°C, transfer the distiller's grain medium to the distiller's grain fermentation tank. The size of the distiller's grain fermentation tank is: 250-300cm long, 180-210cm wide, 120-180cm above the ground, and 120-180cm below the ground. 35~45cm, among which 1~3 days are aerobic fermentation, the distiller's grains are transferred to the pool and not sealed temporarily, 4~10 days are anaerobic fermentation, and the 3rd~5 days of fermentation are to step on the distiller's grains in the pool and seal the pool mouth with clay .

The treatment of distiller's grains biological feed after 9-11 days: after 9-11 days, feed the fermented distiller's grains biological material with full-price compound feed to cattle and sheep, and the ratio of full-price compound feed to distiller's grains biological material is 1:2～ 1:4. The unused distiller's grain biomass is dried at a low temperature of 50-70°C, crushed, and packed in a cool, low-temperature dry place. This feed can be added to the feed of pigs, chickens, and ducks.

Beneficial effects of the present invention : the present invention can convert distiller's grains raw materials into bio-fermented feed integrating microbial cell protein, biologically active small peptide amino acids, functional fatty acids, microbially active probiotics, and compound enzyme preparations. The feed product can not only make up for the amino acids and fatty acids that are easily lacking in conventional feed, but also quickly transform the nutritional components of raw materials that are difficult to absorb, so as to improve the palatability of livestock and enhance digestion, absorption and utilization. It solves the pressure of distiller's grains on environmental pollution, improves the palatability and nutritional content of distiller's grains, solves the problem of human and animal competition for food, provides a specific process for the resource utilization of distiller's grains, and provides a solution for the actual utilization of distiller's grains. It has the following technical advantages and positive effects:

(1) Microbial growth is fast, the cycle is short, the protein content is high, and it is rich in essential amino acids, B vitamins, coenzymes, etc.

(2) Production is not affected by climate conditions and changes in production seasons. The process is simple, low in equipment requirements, low in cost, requires only a small amount of land and labor, low in energy consumption, and effectively utilizes nutrients in distiller's grains.

Description of drawings

Fig. 1 production technique of the present invention;

Detailed ways

Embodiments of the present invention: The bacterial strain combination formula of the production process of processing distiller's grains to produce distiller's grains biological feed of the present invention is: bacterial strains include Bacillus subtilis (Ehrenberg) Cohn), Trichoderma viride (Trichoderma viride), acidophilus Lactobacillus acidophilus, Brettanomyces custersianus; the formula of distiller's grain medium is: fresh distiller's grain 60%, corn flour 10%, rapeseed meal 10%, water 20%, and every 100g distiller's grain culture 15 mg of ferrous sulfate, 12 mg of zinc sulfate, 10 mg of manganese sulfate, 15 mg of copper sulfate and 2 g of calcium hydrogen phosphate were added to the base.

The inoculation amount of Bacillus subtilis was 1.5mL of the bacterial solution per 100g of distiller's grain medium, and the number of viable bacteria was 1.33×10 ¹¹ cfu/mL.

The inoculum amount of Trichoderma viride was 1.5mL of its bacterial solution per 100g of distiller's grain medium, and the number of viable bacteria was 7.0×10 ¹⁰ cfu/mL.

The inoculum amount of Lactobacillus acidophilus is 1.5mL of the bacterial solution per 100g of distiller's grain medium, and the number of viable bacteria is 2.1×10 ¹⁰ cfu/mL.

The inoculum amount of Brettanomyces bantu was 1.5mL of its bacterial solution per 100g of distiller's grain medium, and the number of viable bacteria was 3.5×10 ¹¹ cfu/mL.

As shown in accompanying drawing 1, the production process of processing distiller's grains to produce distiller's grains biological feed includes the following processes: carry out selection of strains and antagonism test to determine whether the selected 4 kinds of bacteria are suitable for mixing and adding without antagonism, expansion of strains, purity and Activity inspection, preparation and sterilization of distiller's grain medium, solid-state fermentation of distiller's grain microorganisms, and treatment of distiller's grain biological feed after 10 days.

Determination of selected strains. The purpose of distiller's grains after fermentation is to make up for the amino acids and fatty acids that are easily lacking in conventional feed, so that the nutrients that are difficult to absorb and other raw materials can be quickly transformed, so as to improve the palatability of livestock and enhance digestion and absorption, that is, dry matter, crude protein, and more The content of unsaturated fatty acids and amino acids increased; the content of crude fiber decreased. Determine the species of choice.

Antagonism test: Group the 4 species of Brettanomyces Bantu, Bacillus subtilis, Trichoderma viride, and Lactobacillus acidophilus in pairs, and inoculate one of the bacteria in each group on the ordinary medium by streaking at intervals of 10mm , and then take another kind of bacteria to draw a straight line vertically, incubate at 30°C for 24 hours, and observe whether the cross point is inhibited and the colony shrinks and disappears. There was no colony atrophy or disappearance at the intersections of strains in each group, and it was concluded that there was no antagonistic effect among the strains, which were suitable for combined fermentation of distiller's grains.

Bacteria expansion culture, purity and activity inspection: inoculate a ring of slant bacteria into 100mL liquid medium for the selected 4 kinds of bacteria, shake the bacteria at 150r/min, 30°C, and expand Bacillus subtilis and Lactobacillus acidophilus after 12h After culturing, they were transferred to fermenters for further expansion. Trichoderma viride and Bantu Brettanomyces were transferred to fermentors for further expansion after 24 hours of expansion. During the expanded culture process, samples were taken from the fermenter for Gram staining and observed under a microscope to determine whether it was the target strain. At the same time, the colony morphology and colony count were observed by coating to determine the activity of the strain.

Preparation and sterilization of distiller's grain medium: distiller's grains are fresh distiller's grains, and when the temperature of the distiller's grains just out of the distillery drops to 80°C, the composition is 60% of fresh distiller's grains, 10% of corn flour, 10% of rapeseed meal, and 20% of water. The percentages here refer to the ratio of each formula to the total weight of the distiller's grain medium; and the ratio of adding 15 mg of ferrous sulfate, 12 mg of zinc sulfate, 10 mg of manganese sulfate, 15 mg of copper sulfate and 2 g of calcium hydrogen phosphate to each 100 g of distiller's grain medium Prepare distiller's grain medium and fully mix other raw materials with it, add lime water to adjust the pH to 6.5, stir and mix well, and stack at normal pressure for sterilization.

Distiller's grain microbial solid-state fermentation: When the temperature of the distiller's grain medium drops to 35°C, inoculate the 4 kinds of bacteria expanded in the fermenter into the distiller's grain medium in equal proportions, with a total inoculation amount of 6%. The viable count of Trichoderma viride reached 7.0×10 ¹⁰ cfu/mL, the viable count of Brettanomyces bantu reached 3.5×10 ¹¹ cfu/mL, and the viable count of Lactobacillus acidophilus reached 2.1×10 ¹⁰ cfu /mL, the number of live bacteria in Bacillus subtilis solution reached 1.33×10 ¹¹ cfu/mL. After inoculation, when the temperature drops to 30°C, transfer the distiller’s grain medium into the distiller’s grain fermentation tank. The size of the distiller’s grain fermentation tank is: 200 cm long, 190 cm wide, 150 cm above the ground, and 40 cm below the ground, of which 1~3 days are Aerobic fermentation, the distiller's grains are transferred to the tank and not sealed temporarily, 4~10 days is anaerobic fermentation, and the 4th day of fermentation is to step on the distiller's grains in the tank and seal the pool mouth with clay for 10 days. After 10 days, the treatment of distiller's grains biological feed: after 10 days, ferment Good distiller's grain biomass is fed to cattle and sheep with full-price compound feed, and the ratio of full-price compound feed to distiller's grain biomass is 1:3. The unused distiller's grains biomass can be dried at 50~70°C at low temperature, crushed, packed, and placed in a cool, low-temperature dry place. This feed can be added to the feed of pigs, chickens, and ducks.

Claims (8)

1. vinasse new bio feed, comprise potale culture medium and bacterial classification, it is characterized in that: the prescription of potale culture medium is: bright distillers ' grains 50%～70%, corn flour 5%～15%, rapeseed dregs 5%～15 %, water 15%～25 %, percentage herein refer to that each prescription accounts for the ratio of the gross weight of potale culture medium; And add in addition ferrous sulfate 12～18 mg, zinc sulfate 10～15 mg, manganese sulfate 8～12 mg, copper sulphate 12～18 mg, calcium monohydrogen phosphate 1～3g in per 80～120 g potale culture medium; Bacterial classification comprises bacillus subtilis, Trichoderma viride, lactobacillus acidophilus and Ban Tu brettanomyce.

2. a kind of vinasse new bio feed according to claim 1, its feature exists: per 80～120 g potale culture medium inoculation bacillus subtilis bacterium liquid, 1～3mL, viable count is 1.0 * 10 ¹¹～1.5 * 10 ¹¹Cfu/mL; Inoculation trichoderma viride bacterium liquid 1～3mL, viable count is 6.8 * 10 ¹⁰～7.2 * 10 ¹⁰Cfu/mL; Inoculating lactobacillus acidophilus bacterium liquid 1～3mL, viable count is 2.0 * 10 ¹⁰～2.2 * 10 ¹⁰Cfu/mL; The figure of inoculation class British mold liquid 1～3mL, viable count is 3.2 * 10 ¹¹～3.6 * 10 ¹¹Cfu/mL.

3. the technological process of a kind of vinasse new bio feed as claimed in claim 1, it is characterized in that: comprise following process: carry out antagonistic effect, bacterial classification enlarges cultivation, purity and active the inspection, the preparation of potale culture medium and sterilization, the processing of vinasse biological feedstuff behind vinasse microorganism solid fermentation and 9～11 d.

4. the production technology of a kind of vinasse new bio feed according to claim 3, it is characterized in that: antagonistic effect: the figure of class brettanomyce, bacillus subtilis, Trichoderma viride, 4 kinds of bacterium of lactobacillus acidophilus are made up grouping in twos, with wherein a kind of bacterium of every group with the interval streak inoculation of 8～12 mm on ordinary culture medium, then getting another bacterium vertically draws a straight line, whether cultivate 22～26 h for 28～32 ℃, observing right-angled intersection place has suppressed and phenomenon generation bacterium colony atrophy and disappearance.

5. the production technology of a kind of vinasse new bio feed according to claim 3, it is characterized in that: bacterial classification enlarges cultivation, purity and active the inspection: selected 4 kinds of bacterium are inoculated respectively a ring inclined-plane bacterium in the 100mL fluid nutrient medium, 150r/min, 30 ℃ shake bacterium, wherein bacillus subtilis, lactobacillus acidophilus enlarge to change over to after cultivating through 10～14h and further enlarge cultivation in the fermentation tank, and Trichoderma viride, the figure of class brettanomyce enlarge to change over to after cultivating through 22～26h and further enlarge cultivation in the fermentation tank; Enlarge in the incubation from fermentation tank sampling and carry out Gram’s staining and observe in microscopically and determine whether to be the purpose bacterial classification, be coated with simultaneously and observe the activity that colonial morphology and clump count are determined bacterial classification.

6. the production technology of a kind of vinasse new bio feed according to claim 3, it is characterized in that: the preparation of potale culture medium and sterilization: by bright distillers ' grains 50%～70%, corn flour 5%～15 %, rapeseed dregs 5%～15 %, water 15%～25 %, percentage herein refers to that each prescription accounts for the ratio of the gross weight of potale culture medium; And add in addition ferrous sulfate 12～18 mg, zinc sulfate 10～15 mg, manganese sulfate 8～12 mg, copper sulphate 12～18 mg, calcium monohydrogen phosphate 1～3g in per 80～120 g potale culture medium, add limewash pH is transferred to 6～7, turn mixing, normal pressure is piled up and is sterilized.

7. the production technology of a kind of vinasse new bio feed according to claim 3, it is characterized in that: the vinasse microorganism solid fermentation: treat when the potale culture medium temperature is down to 32～38 ℃ and will enlarge 4 kinds of bacterium cultivating in the fermentation tank with equal proportion, total inoculum concentration is that 5%～7% ratio is inoculated in the potale culture medium; The bacillus subtilis viable count is 1.0 * 10 ¹¹～1.5 * 10 ¹¹Cfu/mL, trichoderma viride viable count are 6.8 * 10 ¹⁰～7.2 * 10 ¹⁰Cfu/mL, live lactobacillus acidophilus number are 2.0 * 10 ¹⁰～2.2 * 10 ¹⁰Cfu/mL, the figure of class British mold viable count is 3.2 * 10 ¹¹～3.6 * 10 ¹¹Cfu/mL; When treating after the inoculation that temperature is reduced to 28～32 ℃ potale culture medium is changed in the Vinasse fermentation pool, the size of Vinasse fermentation pool is: long 250～300cm, wide 180～210cm, height above ground level is 120～180cm, the below ground height is 35～45cm, and wherein 1～3d is aerobic fermentation, wouldn't seal after vinasse change in the pond, 4～10d days is anaerobic fermentation, and fermentation tracks tramping vinasse in the pond with clay to the 3rd～5d and seals Chi Kou.

8. the production technology of a kind of vinasse new bio feed according to claim 3, it is characterized in that: the processing of vinasse biological feedstuff behind 9～11d: behind 9～11d the biological material collocation of the vinasse that ferment perfect compound feed fed ox, sheep, perfect compound feed is 1:2～1:4 with the biological ratio of expecting of vinasse; It is 50～70 ℃ that the biological material of exhaustless vinasse carries out the low temperature drying temperature, pulverize, package as for shady and cool low temperature drying place, and this feed can be used for adding in the feed of pig, chicken, duck.

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