CN109943504B - Probiotic preparation for harmless treatment of livestock and poultry carcasses and preparation method and application thereof - Google Patents
Probiotic preparation for harmless treatment of livestock and poultry carcasses and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a probiotic preparation for harmless treatment of livestock and poultry carcasses, which is prepared by inoculating bacillus subtilis, bacillus licheniformis, bacillus coagulans and candida utilis after liquid fermentation culture and solid fermentation, drying, crushing and packaging. The invention also discloses a preparation method and application of the probiotic preparation. The invention utilizes the mutual beneficial symbiosis among strains, so that the aspects of the number of the fermented viable bacteria, the enzyme activity, the utilization of macromolecular organic matters and the like are obviously higher than those of single-strain solid-state fermentation, and the invention not only can be used for harmless treatment of livestock and poultry carcasses and deodorization and bacteriostasis of farms, but also can be used for feeding livestock and poultry animals, can promote the digestion and absorption of nutrient substances of feeds, effectively inhibits the growth of pathogenic bacteria, quickly degrades ammonia nitrogen and improves the culture environment.
Description
Technical Field
The invention relates to the technical field of microbial fermentation technology and harmless treatment of dead animal carcasses, in particular to a probiotic preparation for harmless treatment of livestock and poultry carcasses, and a preparation method and application thereof.
Background
The animal husbandry in China is in the period of large-scale, intensive and rapid development, and the number of livestock dead due to natural death or disease is quite large every day. If the harm of natural death is small, and if the carcasses of livestock dead due to diseases are not properly treated, the environment is seriously polluted, and even the health of human beings is threatened. The traditional methods for disposing the animal carcasses mainly comprise a deep burying method, an incineration method, a biodegradation method (fermentation) and the like. However, the deep burying method consumes a large amount of land and has the risk of polluting underground water, soil and the like. The burning method produces toxic gas and pollutes the atmosphere environment. The animal carcasses are treated by microbial fermentation, and if the time is too long, foul smell is generated. Therefore, the animal is treated in a harmless way, and the carcass is fermented and degraded in harmless treatment equipment through microorganisms, so that the carcass treatment time can be shortened, the treatment effect is more thorough, and the animal is safe and environment-friendly. Therefore, a good mixed probiotic preparation is developed to be matched with harmless treatment equipment, so that the deodorization effect is more obvious, and the method is urgent.
The bacillus is a gram-positive bacterium which is strictly aerobic or facultative anaerobic, can synthesize various organic acids, various active enzymes and nutrient metabolites in the propagation process, has the characteristics of high temperature resistance, strong acid and strong alkali resistance, storage resistance, difficult inactivation and the like, and can also quickly decompose macromolecular organic matters and degrade ammonia nitrogen. The strains of Bacillus subtilis, Bacillus licheniformis and Bacillus coagulans used in the invention are all specified in feed additive variety catalog (2013) published by Ministry of agriculture. Wherein the bacillus coagulans is facultative anaerobe, not only has the similar function of lactic acid bacteria, but also can endure the high temperature of 50 ℃.
Yeasts, which are facultative anaerobic microorganisms, can multiply in the digestive tract. The yeast can utilize substances such as ammonia nitrogen, starch, organic acid, hydrogen sulfide and the like as nutrients for self growth, and can propagate in a large quantity to become a dominant flora in competition with the survival of harmful bacteria to inhibit the growth of the harmful bacteria; meanwhile, the cells of the yeast contain abundant protein and vitamins, so the yeast can also be used as high-grade feed for feeding animals. In particular to candida utilis, which is also recorded in feed additive variety catalog (2013) by the Ministry of agriculture.
Disclosure of Invention
The invention aims to provide a probiotic preparation which can be used for harmless treatment of livestock and poultry carcasses, can quickly decompose the livestock carcasses, shorten the carcass treatment time and degrade the odor generated in the treatment process, and has no pollution to the environment.
After intensive research, the inventor of the invention finds that if no probiotic preparation is added when the carcass of the dead livestock is treated by the harmless treatment equipment, the carcass treatment process generates obvious odor and pollutes the environment atmosphere, the carcass treatment time is long, and the time cost and the energy consumption generated therewith are seriously wasted. And the probiotic bacillus can rapidly decompose organic matters, can resist high temperature, is easy to store, is not easy to inactivate and the like.
The probiotic preparation for harmless treatment of the livestock and poultry carcasses is prepared by reasonably matching various aerobic and facultative anaerobic probiotics and adopting a solid fermentation mode combining aerobic and anaerobic fermentation.
The probiotic preparation is prepared by inoculating solid fermentation of bacillus subtilis, bacillus licheniformis, bacillus coagulans and candida utilis after liquid fermentation culture, drying, crushing and packaging.
Preferably, the solid fermentation medium for the probiotic preparation for harmless treatment of livestock and poultry carcasses consists of 10-30 parts by weight of corn protein powder, 20-50 parts by weight of soybean meal, 35-65 parts by weight of rice bran and 2-5 parts by weight of glucose.
Preferably, the solid-state fermentation is prepared by mixing a solid-state fermentation culture medium and the composite probiotic liquid in a solid-liquid ratio of 1:0.8-1(m/v), and fermenting in an aerobic and anaerobic combined mode after uniform mixing.
Preferably, the solid state fermentation temperature is 30-40 ℃, the mixture is stirred and set to be forward turned for 10-30min, stopped for 60-120min, then backward turned for 10-30min, stopped for 60-120min, and the steps are repeated, and the fermentation is finished for 24-48h to obtain the fermentation material to be dried.
Preferably, the viable count of the bacillus subtilis is 5 multiplied by 109CFU/g-10×109CFU/g, viable count of Bacillus licheniformis is 2 × 109CFU/g-7×109CFU/g, viable count of Bacillus coagulans is 2 × 107CFU/g-8×107CFU/g, viable count of Candida utilis 1X 105CFU/g-6×105CFU/g, and water content of 5-9%.
The invention also provides a preparation method of the probiotic preparation for harmless treatment of livestock and poultry carcasses, which comprises the steps of inoculating bacillus subtilis, bacillus licheniformis, bacillus coagulans and candida utilis after liquid fermentation culture for solid state fermentation, drying and crushing after the fermentation is finished, and obtaining the probiotic preparation.
Preferably, the preparation method of the probiotic preparation for harmless treatment of livestock and poultry carcasses comprises the following steps:
(1) activation of original strains: respectively inoculating bacillus subtilis, bacillus licheniformis and bacillus coagulans into a sterilized slant culture medium I to aerobically culture until colonies grow out, and inoculating candida utilis into a sterilized slant culture medium II to aerobically culture until colonies grow out to obtain respective activated slant strains;
the slant culture medium I contains 3g of beef extract, 10g of peptone, 5g of sodium chloride, 20g of agar and 1L of distilled water in proportion, and the pH value is 7.2-7.5; the slant culture medium II contains 20g of glucose, 20g of peptone, 10g of yeast extract, 0.1g of chloramphenicol, 20g of agar powder and 1L of distilled water in proportion, and has natural pH;
preferably, the aerobic culture conditions of the bacillus subtilis, the bacillus licheniformis, the bacillus coagulans and the candida utilis in the step (1) respectively and independently comprise the temperature of 30-40 ℃ and the time of 20-36 h;
(2) preparing a first-level seed solution: respectively inoculating the bacillus subtilis slant strain, the bacillus licheniformis slant strain and the bacillus coagulans slant strain obtained in the step (1) into a sterilized liquid culture medium I, and carrying out aerobic culture at 35-40 ℃ for 20-36h to respectively obtain respective bacillus raw bacteria liquid; inoculating the candida utilis slant strain obtained in the step (1) into a sterilized liquid culture medium II, and carrying out aerobic culture at the temperature of 30-33 ℃ for 24-36h to obtain a yeast raw bacterium liquid;
the liquid culture medium I contains 3g of beef extract, 10g of peptone, 5g of sodium chloride and 1L of distilled water according to a proportion, and the pH value is 7.2-7.5; the liquid culture medium II contains 20g of glucose, 20g of peptone, 10g of yeast extract, 0.1g of chloramphenicol and 1L of distilled water in proportion, and has natural pH;
(3) preparing a secondary seed solution: respectively and independently inoculating the bacillus subtilis, bacillus licheniformis, bacillus coagulans and candida utilis raw bacteria liquid obtained in the step (2) into the same sterilized fermentation medium I according to the inoculation amount of 0.5-2wt%, and carrying out aerobic culture at 30-40 ℃ for 18-36h to obtain composite probiotic liquid;
the fermentation medium I comprises 30g of molasses, 5g of yeast extract, 2g of dipotassium hydrogen phosphate, 0.5g of magnesium sulfate, 0.1g of manganese sulfate and 1L of distilled water according to a proportion, and the pH value is 7.0-7.5;
preferably, the probiotic fermentation in the step (3) is performed under stirring, and the stirring rotation speed is 100-;
(4) solid state fermentation preparation: mixing the solid fermentation culture medium with the compound probiotic liquid obtained in the step (3) according to a solid-liquid ratio of 1:0.8-1(m/v), uniformly mixing, and starting intermittent stirring fermentation;
the components of the solid fermentation medium comprise, by weight, 10-30 parts of corn protein powder, 20-50 parts of soybean meal, 35-65 parts of rice bran and 2-5 parts of glucose;
preferably, in the step (4), the solid state fermentation temperature is 30-40 ℃, the mixture is stirred and set to be forward turned for 10-30min, stopped for 60-120min, then backward turned for 10-30min, stopped for 60-120min, and repeated in this way, and the fermentation is finished for 24-48h to obtain the fermentation material to be dried;
(5) drying and crushing: drying the fermentation material to be dried obtained in the step (4), wherein the drying temperature is 60-100 ℃, the stirring is always in a forward and reverse turning on state in the drying process, the drying time is 24-48h, and the drying is finished and then the crushing treatment is carried out, so that the livestock and poultry carcass harmless treatment probiotic preparation is obtained;
preferably, in the step (5), the probiotic preparation for harmless treatment of livestock and poultry carcasses is in a powder shape, the moisture content is 5-9%, and the viable count of the bacillus subtilis is 5 multiplied by 109CFU/g-10×109CFU/g, viable count of Bacillus licheniformis is 2 × 109CFU/g-7×109CFU/g, viable count of Bacillus coagulans is 2 × 107CFU/g-8×107CFU/g, viable count of Candida utilis 1X 105CFU/g-6×105CFU/g。
The probiotic preparation is applied to harmless treatment of the carcasses of the livestock and poultry.
From the above, it can be seen that the present invention has at least the following advantages:
1. the bacillus coagulans is added, so that the strain not only has the function of lactic acid bacteria, but also has the optimal fermentation growth temperature of 45-50 ℃ and can resist high temperature;
2. according to the invention, aerobic and facultative anaerobic probiotics are adopted, mixed fermentation is carried out through reasonable collocation, and organic matters and ammonia nitrogen can be efficiently degraded through synergistic effect among strains;
3. the mixed bacteria solid state fermentation is adopted, so that the bacterial strain can generate a large amount of metabolites, such as active enzymes, organic acids, bacteriostatic substances and the like, and can accelerate the degradation of the carcass of livestock and inhibit the growth and the propagation of pathogenic bacteria;
4. in the invention, the solid-state fermentation directly adopts the mixed bacteria fermentation liquor to replace the proportion of water for fermentation, so that the solid-state fermentation time can be greatly shortened, sufficient dominant bacteria are added into the fermentation material, and the growth and the propagation of other mixed bacteria can be inhibited;
5. the probiotic preparation for harmless treatment of the livestock and poultry carcasses can quickly degrade the carcasses of dead livestock, remarkably shorten the carcass treatment time, and greatly degrade harmful gases such as ammonia gas, organic amines and the like generated in the carcass treatment process in a short time, thereby achieving the deodorization effect;
6. the probiotic preparation for harmless treatment of the livestock and poultry carcasses has the advantages of safety, environmental protection, high efficiency, no side effect and the like, and the preparation method is simple and can be used for industrial production.
In a word, the invention utilizes the mutual beneficial symbiosis among strains, so that the aspects of the number of the fermented viable bacteria, the enzyme activity, the utilization of macromolecular organic matters and the like are obviously higher than those of single-strain solid-state fermentation, and the invention not only can be used for harmless treatment of livestock and poultry carcasses and deodorization and bacteriostasis of a culture farm, but also can be used for feeding livestock and poultry animals, can promote digestion and absorption of nutrient substances of feed, effectively inhibits the growth of pathogenic bacteria, quickly degrades ammonia nitrogen and improves the culture environment.
Detailed Description
Preferred embodiments of the present invention will be described in more detail below. While the following describes preferred embodiments of the present invention, it should be understood that the present invention may be embodied in various forms and should not be limited by the embodiments set forth herein.
In the following examples and comparative examples:
example 1
This example illustrates a probiotic formulation for the innocent treatment of carcasses of livestock and poultry and a method for preparing the same.
(1) Activation of original strains: respectively inoculating bacillus subtilis, bacillus licheniformis and bacillus coagulans into a sterilized slant culture medium I for aerobic culture until colonies grow out; inoculating candida utilis into a sterilized slant culture medium II for aerobic culture until colonies grow out, and obtaining respective activated slant strains; wherein the slant culture medium I contains 3g of beef extract, 10g of peptone, 5g of sodium chloride, 20g of agar and 1L of distilled water in proportion, and the pH value is 7.2-7.5; the slant culture medium II contains 20g of glucose, 20g of peptone, 10g of yeast extract, 0.1g of chloramphenicol, 20g of agar powder and 1L of distilled water in proportion, and has natural pH; respectively and independently carrying out aerobic culture on the bacillus subtilis, the bacillus licheniformis and the bacillus coagulans at 37 ℃ for 24 hours, and carrying out aerobic culture on candida utilis at 33 ℃ for 36 hours;
(2) preparing a first-level seed solution: respectively inoculating the bacillus subtilis slant strain, the bacillus licheniformis slant strain and the bacillus coagulans slant strain obtained in the step (1) into a sterilized liquid culture medium I, and carrying out aerobic culture at 37 ℃ for 24h to respectively obtain respective bacillus raw bacteria liquid; inoculating the candida utilis slant strain obtained in the step (1) into a sterilized liquid culture medium II, and carrying out aerobic culture at 33 ℃ for 36h to obtain a yeast raw bacterium liquid; wherein the liquid culture medium I contains 3g of beef extract, 10g of peptone, 5g of sodium chloride and 1L of distilled water according to a certain proportion, and the pH value is 7.2-7.5; the liquid culture medium II contains 20g of glucose, 20g of peptone, 10g of yeast extract, 0.1g of chloramphenicol and 1L of distilled water in proportion, and has natural pH;
(3) preparing a secondary seed solution: respectively and independently inoculating the bacillus subtilis, bacillus licheniformis, bacillus coagulans and candida utilis raw bacterium liquid obtained in the step (2) into the same sterilized fermentation medium I according to the inoculation amount of 1 wt%, and carrying out aerobic culture at 37 ℃ for 36h, wherein the culture conditions are as follows: stirring is carried out under the condition of a rotating speed of 180r/min to obtain the composite probiotic liquid; wherein the fermentation medium I comprises 30g of molasses, 5g of yeast extract, 2g of dipotassium hydrogen phosphate, 0.5g of magnesium sulfate, 0.1g of manganese sulfate and 1L of distilled water according to a proportion, and the pH value is 7.0-7.5;
(4) solid state fermentation preparation: uniformly mixing the compound probiotic liquid obtained in the step (3) with a solid-liquid fermentation culture medium according to a solid-liquid ratio of 0.8:1, setting the fermentation temperature to 37 ℃, stirring, positively turning over for 10min, stopping for 60min, reversely turning over for 10min, stopping for 60min, repeating the steps, ending the fermentation for 24h, adjusting the temperature to 60 ℃, starting and drying for 48h, crushing and packaging after the drying is ended, and thus obtaining the livestock and poultry carcass harmless treatment probiotic preparation; wherein the components of the solid fermentation medium comprise 13 parts of corn protein powder, 20 parts of soybean meal, 65 parts of rice bran and 2 parts of glucose according to parts by weight, the obtained probiotic preparation is powdery, the water content is 6 percent, and the viable count of the bacillus subtilis is 5.8 multiplied by 109CFU/g, viable count of Bacillus licheniformis is 5.1 × 109CFU/g, viable count of Bacillus coagulans is 4.2 × 107CFU/g, viable count of Candida utilis 2.4X 105CFU/g。
Example 2
This example illustrates a probiotic formulation for the innocent treatment of carcasses of livestock and poultry and a method for preparing the same.
(1) The preparation method of the primary strain activated to the secondary seed liquid is the same as that of example 1, and is not repeated.
(2) Solid state fermentation preparation: and uniformly mixing the composite probiotic liquid and the solid-state fermentation culture medium according to the solid-liquid ratio of 1:1, and setting the fermentation temperature to be 37 ℃. And (3) stirring, positively overturning for 30min, stopping for 60min, reversely overturning for 30min, stopping for 60min, repeating the steps, finishing fermentation for 48h, adjusting the temperature to 80 ℃, starting and drying for 24h, crushing and packaging after drying, and thus obtaining the livestock and poultry carcass harmless treatment probiotic preparation. The solid fermentation medium comprises, by weight, 20 parts of corn protein powder, 20 parts of soybean meal powder, 60 parts of rice bran and 3 parts of glucose. Wherein the probiotic preparation is in the form of powder, water7 percent of the total weight of the bacillus subtilis, and the viable count of the bacillus subtilis is 9.5 multiplied by 109CFU/g, viable count of Bacillus licheniformis is 6.4 × 109CFU/g, viable count of Bacillus coagulans of 7.1 × 107CFU/g, viable count of Candida utilis 5.6 × 105CFU/g。
Example 3
This example illustrates a probiotic formulation for the innocent treatment of carcasses of livestock and poultry and a method for preparing the same.
(1) The preparation method of the primary strain activated to the secondary seed liquid is the same as that of example 1, and is not repeated.
(2) Solid state fermentation preparation: and uniformly mixing the composite probiotic liquid and the solid-state fermentation culture medium according to the solid-liquid ratio of 1:1, and setting the fermentation temperature to be 37 ℃. And (3) stirring, positively overturning for 30min, stopping for 120min, reversely overturning for 30min, stopping for 120min, repeating the steps, finishing fermentation for 48h, adjusting the temperature to 80 ℃, starting and drying for 48h, crushing and packaging after drying, and thus obtaining the livestock and poultry carcass harmless treatment probiotic preparation. The solid fermentation medium comprises, by weight, 30 parts of corn protein powder, 30 parts of soybean meal, 40 parts of rice bran and 5 parts of glucose. Wherein the obtained probiotic preparation is in powder form, has water content of 6%, and viable count of Bacillus subtilis of 6.5 × 109CFU/g, viable count of Bacillus licheniformis is 4.7 × 109CFU/g, viable count of Bacillus coagulans is 3.3 × 107CFU/g, viable count of Candida utilis 2.6 × 105CFU/g。
Comparative example 1
The comparative example is used to illustrate a reference probiotic preparation for innocent treatment of carcasses of livestock and poultry and a preparation method thereof.
A livestock and poultry carcass harmless treatment probiotic preparation was prepared according to the method of example 2, except that no bacillus subtilis was used in this comparative example, and accordingly, the preparation process of a livestock and poultry carcass harmless treatment probiotic preparation did not include the steps of activating bacillus subtilis and preparing a primary seed solution corresponding to bacillus subtilis, and no bacillus subtilis was used in the preparation process of the secondary seed solution, to obtain a livestock and poultry carcass harmless treatment probiotic preparation.
Comparative example 2
The comparative example is used to illustrate a reference probiotic preparation for innocent treatment of carcasses of livestock and poultry and a preparation method thereof.
A livestock and poultry carcass harmless treatment probiotic preparation was prepared according to the method of example 2, except that bacillus coagulans was not used in this comparative example, and accordingly, the preparation process of a livestock and poultry carcass harmless treatment probiotic preparation did not include the steps of bacillus coagulans activation and preparation of primary seed liquid corresponding to bacillus coagulans, and no bacillus coagulans was contained in the preparation process of secondary seed liquid, resulting in a livestock and poultry carcass harmless treatment probiotic preparation.
Comparative example 3
The comparative example is used to illustrate a reference probiotic preparation for innocent treatment of carcasses of livestock and poultry and a preparation method thereof.
A livestock and poultry carcass innocent treatment probiotic preparation was prepared according to the method of example 2, except that the comparative example did not employ candida utilis, and accordingly, the preparation process of the livestock and poultry carcass innocent treatment probiotic preparation did not include the preparation steps of activation of candida utilis and the primary seed solution corresponding to the candida utilis, and no candida utilis was used in the preparation process of the secondary seed solution, and a livestock and poultry carcass innocent treatment probiotic preparation was obtained.
Test example
The test method comprises the following steps: in a certain pig farm, seven groups of tests were set, three of which were set as test groups and the other four were set as control groups, and the livestock and poultry carcass innocent treatment probiotic preparation obtained in examples 1 to 3 of the present invention was used in the test groups. And the reference livestock carcass harmless treatment probiotic preparation obtained in the comparative examples 1-3 is respectively used in the four groups of control groups, and the livestock carcass harmless treatment probiotic preparation is not used in the other group (as a blank control). In each group of experiments, 10 pigs dead of illness (about 1 ton) in the same time period are selected and put into harmless treatment equipment (existing equipment in the market). The six groups of test groups are added with the livestock and poultry carcass harmless treatment probiotic preparation according to the amount of 2kg/1 ton of live pigs. The blank control is added with a non-probiotic carrier preparation (corn protein powder, soybean meal and rice bran in a ratio of 2:2:6) according to the amount of 2kg/1 ton of live pigs. After the temperature of the harmless treatment equipment is increased from room temperature to 80 ℃ within 3 hours in a short time, the temperature in the equipment is maintained within the range of 80-100 ℃ for a long time, and a stirring system continuously operates during the operation of the equipment. And (3) during the period of processing the dead pig carcass for 3h, paying attention to whether ammonia odor is generated or not and detecting pathogenic bacteria of the sample after the processing is finished.
Note: "+" indicates ammonia odor generation, more "+" indicates more concentration; "-" indicates no ammonia odor generation
The test result shows that: the three test groups hardly or slightly generate odor and ammonia gas within 3 hours after the dead pig bodies are treated, and the deodorization effect is obvious. Among the three comparative examples, comparative example 2 in which no Bacillus coagulans was added had the least ammonia odor removal effect, and thus, Bacillus coagulans was found to have a certain effect.
The six treatment groups have the following effects after the dead pigs are treated: the pig feed is basically black brown powder in appearance, and no obvious viscera, organs and muscle tissues of the live pigs can be seen, wherein small bones are completely degraded, large bones are in small blocks and are seriously corroded. The blank control group has undesirable effects, most of the blank control group is not decomposed completely, the fur bones can be seen, and at least one additional treatment day is needed if the appearance of the treatment group is to be achieved.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the various features described in the above embodiments may be combined in any suitable manner without departing from the scope of the invention. The invention is not described in detail in order to avoid unnecessary repetition.
Claims (6)
1. A probiotic preparation for harmless treatment of livestock and poultry carcasses is characterized in that: the bacillus subtilis, the bacillus licheniformis, the bacillus coagulans and the candida utilis are subjected to liquid state fermentation culture and then inoculated with solid state fermentation, and the bacillus coagulans is prepared after drying, crushing and packaging;
the solid fermentation is prepared by mixing a solid fermentation culture medium and compound probiotic liquid according to a solid-liquid ratio of 1:0.8-1(m/v), and fermenting in a mode of combining aerobic fermentation and anaerobic fermentation after uniform mixing;
the viable count of the bacillus subtilis is 5 multiplied by 109CFU/g-10×109CFU/g, viable count of Bacillus licheniformis is 2 × 109CFU/g-7×109CFU/g, viable count of Bacillus coagulans is 2 × 107CFU/g-8×107CFU/g, viable count of Candida utilis 1X 105CFU/g-6×105CFU/g, moisture content 5-9%;
and the solid state fermentation temperature is 30-40 ℃, the mixture is uniformly mixed, stirred, positively turned for 10-30min, stopped for 60-120min, reversely turned for 10-30min and stopped for 60-120min, the steps are repeated, and the fermentation is finished for 24-48h to obtain the fermentation material to be dried.
2. The probiotic preparation for harmless treatment of livestock and poultry carcasses according to claim 1, characterized in that: the solid fermentation medium comprises, by weight, 10-30 parts of corn protein powder, 20-50 parts of soybean meal, 35-65 parts of rice bran and 2-5 parts of glucose.
3. The method for preparing the probiotic preparation for harmless treatment of livestock and poultry carcasses according to claim 1, which is characterized by comprising the following steps of: the preparation method comprises the following steps:
(1) activation of original strains: respectively inoculating bacillus subtilis, bacillus licheniformis and bacillus coagulans into a sterilized slant culture medium I to aerobically culture until colonies grow out, and inoculating candida utilis into a sterilized slant culture medium II to aerobically culture until colonies grow out to obtain respective activated slant strains;
the slant culture medium I contains 3g of beef extract, 10g of peptone, 5g of sodium chloride, 20g of agar and 1L of distilled water in proportion, and the pH value is 7.2-7.5; the slant culture medium II contains 20g of glucose, 20g of peptone, 10g of yeast extract, 0.1g of chloramphenicol, 20g of agar powder and 1L of distilled water in proportion, and has natural pH;
(2) preparing a first-level seed solution: respectively inoculating the bacillus subtilis slant strain, the bacillus licheniformis slant strain and the bacillus coagulans slant strain obtained in the step (1) into a sterilized liquid culture medium I, and carrying out aerobic culture at 35-40 ℃ for 20-36h to respectively obtain respective bacillus raw bacteria liquid; inoculating the candida utilis slant strain obtained in the step (1) into a sterilized liquid culture medium II, and carrying out aerobic culture at the temperature of 30-33 ℃ for 24-36h to obtain a yeast raw bacterium liquid;
the liquid culture medium I contains 3g of beef extract, 10g of peptone, 5g of sodium chloride and 1L of distilled water according to a proportion, and the pH value is 7.2-7.5; the liquid culture medium II contains 20g of glucose, 20g of peptone, 10g of yeast extract, 0.1g of chloramphenicol and 1L of distilled water in proportion, and has natural pH;
(3) preparing a secondary seed solution: respectively and independently inoculating the bacillus subtilis, bacillus licheniformis, bacillus coagulans and candida utilis raw bacteria liquid obtained in the step (2) into the same sterilized fermentation medium I according to the inoculation amount of 0.5-2wt%, and carrying out aerobic culture at 30-40 ℃ for 18-36h to obtain composite probiotic liquid;
the fermentation medium I comprises 30g of molasses, 5g of yeast extract, 2g of dipotassium hydrogen phosphate, 0.5g of magnesium sulfate, 0.1g of manganese sulfate and 1L of distilled water according to a proportion, and the pH value is 7.0-7.5;
(4) solid state fermentation preparation: mixing the solid fermentation culture medium with the compound probiotic liquid obtained in the step (3) according to a solid-liquid ratio of 1:0.8-1(m/v), uniformly mixing, and starting intermittent stirring fermentation;
the components of the solid fermentation medium comprise, by weight, 10-30 parts of corn protein powder, 20-50 parts of soybean meal, 35-65 parts of rice bran and 2-5 parts of glucose;
the solid state fermentation temperature is 30-40 ℃, the mixture is stirred and set to be forward turned for 10-30min, stopped for 60-120min, then backward turned for 10-30min, stopped for 60-120min, the steps are repeated, and the fermentation is finished for 24-48h to obtain a fermentation material to be dried;
(5) drying and crushing: drying the fermentation material to be dried obtained in the step (4), wherein the drying temperature is 60-100 ℃, the stirring is always in a forward and reverse turning on state in the drying process, the drying time is 24-48h, and the drying is finished and then the crushing treatment is carried out, so that the livestock and poultry carcass harmless treatment probiotic preparation is obtained;
the probiotic preparation for harmless treatment of livestock and poultry carcasses is in a powder shape, the water content is 5-9%, and the viable count of bacillus subtilis is 5 multiplied by 109CFU/g-10×109CFU/g, viable count of Bacillus licheniformis is 2 × 109CFU/g-7×109CFU/g, viable count of Bacillus coagulans is 2 × 107CFU/g-8×107CFU/g, viable count of Candida utilis 1X 105CFU/g-6×105CFU/g。
4. The method for preparing the probiotic preparation for harmless treatment of livestock and poultry carcasses according to claim 3, wherein the method comprises the following steps: in the step (1), aerobic culture conditions of the bacillus subtilis, the bacillus licheniformis, the bacillus coagulans and the candida utilis are as follows: the temperature is 30-40 ℃, and the time is 20-36 h.
5. The method for preparing the probiotic preparation for harmless treatment of livestock and poultry carcasses according to claim 3, wherein the method comprises the following steps: in the step (3), the probiotic fermentation is performed under stirring, and the stirring speed is 100-180 r/min.
6. Use of a livestock and poultry carcass innocent treatment probiotic preparation according to any one of claims 1-5 for removing ammonia odor of livestock and poultry carcasses.
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| CN106282038A (en) * | 2016-08-05 | 2017-01-04 | 湖北凌卓生物工程有限公司 | A kind of composite efficient microbial deodorant and preparation method thereof |
| EP3187579A1 (en) * | 2014-08-25 | 2017-07-05 | Japan Environmental Science Company | Composition, support, wastewater treatment system, wastewater treating method, deodorization method, and batch wastewater treating method |
| CN109251874A (en) * | 2018-09-25 | 2019-01-22 | 厦门惠盈动物科技有限公司 | A kind of probiotics preparation and its preparation method and application |
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| EP3187579A1 (en) * | 2014-08-25 | 2017-07-05 | Japan Environmental Science Company | Composition, support, wastewater treatment system, wastewater treating method, deodorization method, and batch wastewater treating method |
| CN106282038A (en) * | 2016-08-05 | 2017-01-04 | 湖北凌卓生物工程有限公司 | A kind of composite efficient microbial deodorant and preparation method thereof |
| CN109251874A (en) * | 2018-09-25 | 2019-01-22 | 厦门惠盈动物科技有限公司 | A kind of probiotics preparation and its preparation method and application |
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