CN103783321B - Shelled Turtle Trionyx Sinensis probiotic feed additive and preparation method - Google Patents
Shelled Turtle Trionyx Sinensis probiotic feed additive and preparation method Download PDFInfo
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Abstract
The present invention relates to a kind of Shelled Turtle Trionyx Sinensis probiotic feed additive.Primarily of probio, carrier inserts composition, described carrier inserts is sterilizing wheat bran; Described probio is bacillus subtilis, bafillus natto, Lactobacillus casei and candida utili bacterium, and four kinds of bacterial classifications are added on obtained bacterium powder in sterile carrier inserts by the adding proportion of 0.4 ~ 0.6:0.4 ~ 0.6:0.8 ~ 1.2:0.8 ~ 1.2.The present invention can improve Shelled Turtle Trionyx Sinensis growth performance, reduce feed coefficient, improve immunologic function and reduce disease, also have low cost, pollution-free, without the advantage such as the resistance to the action of a drug and noresidue, be a kind of feed addictive of environmental protection.
Description
Technical field
The invention belongs to probiotics and field of feed additive technology, be specifically related to Shelled Turtle Trionyx Sinensis probiotic feed additive and preparation method.
Background technology
Along with the continuous expansion of the market demand, Shelled Turtle Trionyx Sinensis industrial aquaculture develops rapidly, but the intensive breeding way of high density changes their natural life habit, causes weakening of its resistance against diseases, disease is increasing, have impact on further developing of Shelled Turtle Trionyx Sinensis aquaculture.In modern animal husbandry, add antibiotic or chemical synthetic drug in feed, to disease preventing and treating, growth promoting effects, saves feed, improves fanning economics and plays an important role.But Long-Time Service or excessive use can cause fish to produce immunosupress, medicament residue and pathogen and produce the side effects such as drug resistance, cause aquatic products quality to reduce, constitute a threat to food security, and can cause bad impact to environment and human health.International linked groups has defined some antibiotic and the application of chemicals on aquatic products.China forbids in feed, add antibiotic and chemical synthetic drug is also an inexorable trend, makes great efforts the new way of seeking green health cultivation.
At present, nutrition and health are the focuses that people pay close attention to, and along with the raising of people's living standard, people get more and more to the demand of pollution-free food.For this reason, domestic and international scientific research personnel begin one's study and apply nontoxic, pollution-free, without the resistance to the action of a drug, noresidue, there is prevention Shelled Turtle Trionyx Sinensis disease and improve the probio of its production performance.Probio is the bacterium useful to humans and animals, and probio, as the research of feed addictive and extensive use, brings brand-new situation by for feed addictive, brings huge economic benefit to culture fishery.
Summary of the invention
The object of the present invention is to provide a kind of NEW TYPE OF COMPOSITE probiotic feed additive, this feed addictive has regulating intestinal canal colony balance, growth promotion, enhancing resistance against diseases and the several functions such as to improve food conversion ratio, and low cost, pollution-free, noresidue, does not develop immunity to drugs.
In order to solve the problems of the technologies described above, the technical solution used in the present invention is:
Described prebiotic feed addictive is made up of the bacterium powder of bacillus subtilis, bafillus natto, Lactobacillus casei, candida utili bacterium, the bacterium powder of bacillus subtilis, bafillus natto, Lactobacillus casei, candida utili bacterium is obtained using the wheat bran of sterilizing as absorption carrier respectively, and its constituent mass ratio is: 0.4 ~ 0.6:0.4 ~ 0.6:0.8 ~ 1.2:0.8 ~ 1.2.The viable count of bacterium powder is bacillus subtilis 1 × 10
8cFU/g, bafillus natto 1 × 10
9cFU/g, Lactobacillus casei 1 × 10
8or 10
11cFU/g, candida utili bacterium 1 × 10
8cFU/g.
Preparation method and the step of Shelled Turtle Trionyx Sinensis probiotic feed additive are as follows:
(1) bacillus subtilis (numbering: CICC21746, the Bacillus subtilis) preparation of preparation
The solid ordinary culture medium activation of the bacillus subtilis sterilizing of first inclined-plane being preserved, through 30 DEG C ~ 37 DEG C activation culture 24h ~ 36h, then on the liquid ordinary culture medium of sterilizing, by the bacillus subtilis of 10% inoculum concentration access activation, temperature 30 DEG C, rotating speed 180r/min, pressure 0.05MPa, cultivate 24h ~ 36h, bacterium number bacillus subtilis 1 × 10
8cFU/g, can obtain liquid bacillus subtilis formulation.
Consisting of of described solid ordinary culture medium nutritional labeling: 5 parts, sodium chloride, beef extract 3 parts, peptone 10 parts, 20 parts, agar, distilled water 1000 parts, pH7.2 ~ 7.4.Sterilising temp 115 DEG C, sterilization time 20min.Quality by gram.
Consisting of of described liquid ordinary culture medium nutritional labeling: 5 parts, sodium chloride, beef extract 3 parts, peptone 10 parts, distilled water 1000 parts, pH7.2 ~ 7.4.Sterilising temp 115 DEG C, sterilization time 20min.
Mixed with water by wheat bran, the two mass ratio is 1:0.5, and high-temperature sterilization cools, and obtains sterile carrier inserts.
By liquid bacillus subtilis formulation and sterile carrier inserts Homogeneous phase mixing, the two weight ratio is 1:10, and 30 DEG C ~ 37 DEG C ventilating fermentation 24h ~ 36h post-dryings are pulverized, and can obtain bacillus subtilis bacterium powder, the viable count of bacterium powder is more than or equal to 1 × 10
8cFU/g.
(2) bafillus natto (numbering: CICC20443, the Bacillus natto) preparation of preparation
The solid Sha Baolao culture medium activation of the bafillus natto sterilizing of first inclined-plane being preserved, through 30 DEG C ~ 37 DEG C activation culture 24h ~ 36h, then on the liquid Sha Baolao culture medium of sterilizing, by the bafillus natto of 10% inoculum concentration access activation, temperature 37 DEG C, rotating speed 180r/min, pressure 0.05MPa, cultivate 24h ~ 36h, liquid natto bacillus preparation can be obtained.
Consisting of of described solid Sha Baolao medium nutrient content: peptone 10 parts, maltose 40 parts, 20 parts, agar, distilled water 1000 parts, pH6.8, sterilising temp 115 DEG C, sterilization time 20min.
Consisting of of described liquid Sha Baolao medium nutrient content: peptone 10 parts, maltose 40 parts, distilled water 1000 parts, Ph6.8.Sterilising temp 115 DEG C, sterilization time 20min.
Mixed with water by wheat bran, the two mass ratio is 1:0.5, and high-temperature sterilization cools, and obtains sterile carrier inserts.
By liquid natto bacillus preparation and sterile carrier inserts Homogeneous phase mixing, the two weight ratio is 1:10, and 30 DEG C ~ 37 DEG C ventilating fermentation 24h ~ 36h post-dryings are pulverized, and can obtain bafillus natto bacterium powder, bafillus natto viable count is more than or equal to 1 × 10
9cFU/g.
(3) Lactobacillus casei (numbering: the preparation of CICC6105, Lactobacillus casei preparation
The solid MRS culture medium activation of the Lactobacillus casei sterilizing of first inclined-plane being preserved, through 30 DEG C ~ 37 DEG C activation culture 24h ~ 36h, then on the liquid MRS culture medium of sterilizing, by the Lactobacillus casei of 10% inoculum concentration access activation, temperature 37 DEG C, quiescent culture 24h ~ 36h, can obtain liquid Lactobacillus casei preparation.
Consisting of of described solid MRS medium nutrient content: peptone 10 parts, beef extract 10 parts, yeast extract 5 parts, diammonium hydrogen citrate [(NH
4)
2hC
6h
5o
7] 2 parts, glucose (C
6h
12o
6h
2o) 20 parts, Tween 80 1 part, sodium acetate (CH
3cOONa3H
2o) 5g, dipotassium hydrogen phosphate (K
2hPO
43H
2o) 2 parts, magnesium sulfate (MgSO
47H
2o) 0.58 part, manganese sulfate (MnSO
4h
2o) 0.25 part, 18 parts, agar, distilled water 1000 parts, pH:6.2 ~ 6.6.Sterilising temp 121 DEG C, sterilization time 15min.
Consisting of of described liquid MRS medium nutrient content: peptone 10 parts, beef extract 10 parts, yeast extract 5 parts, diammonium hydrogen citrate [(NH
4)
2hC
6h
5o
7] 2 parts, glucose (C
6h
12o
6h
2o) 20 parts, Tween 80 1 part, sodium acetate (CH
3cOONa3H
2o) 5 parts, dipotassium hydrogen phosphate (K
2hPO
43H
2o) 2 parts, magnesium sulfate (MgSO
47H
2o) 0.58 part, manganese sulfate (MnSO
4h
2o) 0.25 part, distilled water 1000 parts, pH6.2 ~ 6.6, sterilising temp 121 DEG C, sterilization time 15min.
Mixed with water by wheat bran, the two mass ratio is 1:0.5, and high-temperature sterilization cools, and obtains sterile carrier inserts.
By liquid Lactobacillus casei preparation and sterile carrier inserts Homogeneous phase mixing, the two weight ratio is 1:10,30 DEG C ~ 37 DEG C sealing and fermenting 24h ~ 36h post-dryings are pulverized, and can obtain Lactobacillus casei bacterium powder, Lactobacillus casei powder viable count is more than or equal to Lactobacillus casei 1 × 10
8or 10
11cFU/g.
(4) candida utili bacterium (numbering: CICC1314, the Candida utilis) preparation of preparation
The solid Sha Baolao culture medium activation of the candida utili bacterium sterilizing of first inclined-plane being preserved, through 28 DEG C ~ 30 DEG C activation culture 24h ~ 36h, then on the liquid Sha Baolao culture medium of sterilizing, by the candida utili bacterium of 10% inoculum concentration access activation, temperature 30 DEG C, rotating speed 180r/min, pressure 0.05MPa, cultivate 24h ~ 36h, liquid candida utili bacteria preparation can be obtained.
Consisting of of described solid Sha Baolao medium nutrient content: peptone 10 parts, glucose 40 parts, 20 parts, agar, distilled water 1000 parts, pH6.8.Sterilising temp 115 DEG C, sterilization time 20min.
Consisting of of described liquid Sha Baolao medium nutrient content: peptone 10 parts, glucose 40 parts, distilled water 1000 parts, pH6.8.Sterilising temp 115 DEG C, sterilization time 20min.
Mixed with water by wheat bran, the two mass ratio is 1:0.5, and high-temperature sterilization cools, and obtains sterile carrier inserts.
By liquid candida utili bacteria preparation and sterile carrier inserts Homogeneous phase mixing, the two weight ratio is 1:10, and 30 DEG C ~ 37 DEG C fermentation 24h ~ 36h post-dryings are pulverized, and can obtain candida utili bacterium bacterium powder, candida utili bacterium viable count is more than or equal to 1 × 10
8cFU/g.
(5) preparation of composite probiotics preparations
Above-mentioned bacillus subtilis bacterium powder, bafillus natto bacterium powder, Lactobacillus casei bacterium powder, candida utili bacterium bacterium powder are mixed by the ratio uniform of 0.4 ~ 0.6:0.4 ~ 0.6:0.8 ~ 1.2:0.8 ~ 1.2, obtains composite probiotics preparations powder form.
Described composite probiotics preparations can add in Shelled Turtle Trionyx Sinensis feed with the dosage of 1000mg/kg and uses.
All complete with stainless steel fermentation tank in fermentation production process.
Above-mentioned bacterial classification can be buied from " Chinese industrial Microbiological Culture Collection administrative center (CICC) ": bacillus subtilis (numbering: 21746, Bacillus subtilis), bafillus natto (numbering: 20443, Bacillus natto), candida utili bacterium (numbering: 1314, Candida utilis), Lactobacillus casei (numbering: 6105, Lactobacillus casei).
Above-mentioned Nostoc commune Vanch based component, Sha Baolao medium component, MRS medium component and inserts wheat bran are commercially available.
The present invention has following beneficial effect:
(1) improve feed quality, improve feed nutrition, strengthen absorbing feed, promote Shelled Turtle Trionyx Sinensis growth.
(2) enhanced machine body immunity function, improves diseases prevention ability, reduces disease and occurs.
Detailed description of the invention
Prepared by embodiment 1 Shelled Turtle Trionyx Sinensis composite probiotics feed additive
Composite probiotics feed additive, comprises probio and absorption carrier inserts, and probio comprises bacillus subtilis, bafillus natto, candida utili bacterium and Lactobacillus casei, and absorption carrier inserts is sterilizing wheat bran.It is 0.5:0.5:1:1 at composite probiotics feed additive mass ratio (in every gram of additive each bacterium powder amount ratio).In the feed ration of basis, composite probiotics preparations powder-type addition is according to 1000mg ~ 2000mg/(kg basal diet) standard interpolation.
The preparation method of above-mentioned composite probiotics feed additive is:
The first step: probiotics fermention is cultivated.
(1) bacillus subtilis (numbering: CICC21746, the Bacillus subtilis) preparation of preparation
The solid ordinary culture medium activation of the bacillus subtilis sterilizing of first inclined-plane being preserved, through 30 DEG C ~ 37 DEG C activation culture 24h ~ 36h, then on the liquid ordinary culture medium of sterilizing, by the bacillus subtilis of 10% inoculum concentration access activation, temperature 30 DEG C, rotating speed 180r/min, pressure 0.05MPa, cultivate 24h ~ 36h, liquid bacillus subtilis formulation can be obtained.
Consisting of of described solid ordinary culture medium nutritional labeling: 5 parts, sodium chloride, beef extract 3 parts, peptone 10 parts, 20 parts, agar, distilled water 1000 parts, pH7.2 ~ 7.4.Sterilising temp 115 DEG C, sterilization time 20min.Quality by gram.
Consisting of of described liquid ordinary culture medium nutritional labeling: 5 parts, sodium chloride, beef extract 3 parts, peptone 10 parts, distilled water 1000 parts, pH7.2 ~ 7.4.Sterilising temp 115 DEG C, sterilization time 20min.
Mixed with water by wheat bran, the two mass ratio is 1:0.5, and high-temperature sterilization cools, and obtains sterile carrier inserts.
By liquid bacillus subtilis formulation and sterile carrier inserts Homogeneous phase mixing, the two weight ratio is 1:10, and 30 DEG C ~ 37 DEG C ventilating fermentation 24h ~ 36h post-dryings are pulverized, and can obtain bacillus subtilis bacterium powder.
(2) bafillus natto (numbering: CICC20443, the Bacillus natto) preparation of preparation
The solid Sha Baolao culture medium activation of the bafillus natto sterilizing of first inclined-plane being preserved, through 30 DEG C ~ 37 DEG C activation culture 24h ~ 36h, then on the liquid Sha Baolao culture medium of sterilizing, by the bafillus natto of 10% inoculum concentration access activation, temperature 37 DEG C, rotating speed 180r/min, pressure 0.05MPa, cultivate 24h ~ 36h, liquid natto bacillus preparation can be obtained.
Consisting of of described solid Sha Baolao medium nutrient content: peptone 10 parts, maltose 40 parts, 20 parts, agar, distilled water 1000 parts, pH6.8, sterilising temp 115 DEG C, sterilization time 20min.
Consisting of of described liquid Sha Baolao medium nutrient content: peptone 10 parts, maltose 40 parts, distilled water 1000 parts, Ph6.8.Sterilising temp 115 DEG C, sterilization time 20min.
Mixed with water by wheat bran, the two mass ratio is 1:0.5, and high-temperature sterilization cools, and obtains sterile carrier inserts.
By liquid natto bacillus preparation and sterile carrier inserts Homogeneous phase mixing, the two weight ratio is 1:10, and 30 DEG C ~ 37 DEG C ventilating fermentation 24h ~ 36h post-dryings are pulverized, and can obtain bafillus natto bacterium powder.
(3) Lactobacillus casei (numbering: the preparation of CICC6105, Lactobacillus casei preparation
The solid MRS culture medium activation of the Lactobacillus casei sterilizing of first inclined-plane being preserved, through 30 DEG C ~ 37 DEG C activation culture 24h ~ 36h, then on the liquid MRS culture medium of sterilizing, by the Lactobacillus casei of 10% inoculum concentration access activation, temperature 37 DEG C, quiescent culture 24h ~ 36h, can obtain liquid Lactobacillus casei preparation.
Consisting of of described solid MRS medium nutrient content: peptone 10 parts, beef extract 10 parts, yeast extract 5 parts, diammonium hydrogen citrate [(NH
4)
2hC
6h
5o
7] 2 parts, glucose (C
6h
12o
6h
2o) 20 parts, Tween 80 1 part, sodium acetate (CH
3cOONa3H
2o) 5g, dipotassium hydrogen phosphate (K
2hPO
43H
2o) 2 parts, magnesium sulfate (MgSO
47H
2o) 0.58 part, manganese sulfate (MnSO
4h
2o) 0.25 part, 18 parts, agar, distilled water 1000 parts, pH:6.2 ~ 6.6.Sterilising temp 121 DEG C, sterilization time 15min.
Consisting of of described liquid MRS medium nutrient content: peptone 10 parts, beef extract 10 parts, yeast extract 5 parts, diammonium hydrogen citrate [(NH
4)
2hC
6h
5o
7] 2 parts, glucose (C
6h
12o
6h
2o) 20 parts, Tween 80 1 part, sodium acetate (CH
3cOONa3H
2o) 5 parts, dipotassium hydrogen phosphate (K
2hPO
43H
2o) 2 parts, magnesium sulfate (MgSO
47H
2o) 0.58 part, manganese sulfate (MnSO
4h
2o) 0.25 part, distilled water 1000 parts, pH6.2 ~ 6.6, sterilising temp 121 DEG C, sterilization time 15min.
Mixed with water by wheat bran, the two mass ratio is 1:0.5, and high-temperature sterilization cools, and obtains sterile carrier inserts.
By liquid Lactobacillus casei preparation and sterile carrier inserts Homogeneous phase mixing, the two weight ratio is 1:10, and 30 DEG C ~ 37 DEG C sealing and fermenting 24h ~ 36h post-dryings are pulverized, and can obtain Lactobacillus casei bacterium powder.
(4) candida utili bacterium (numbering: CICC1314, the Candida utilis) preparation of preparation
The solid Sha Baolao culture medium activation of the candida utili bacterium sterilizing of first inclined-plane being preserved, through 28 DEG C ~ 30 DEG C activation culture 24h ~ 36h, then on the liquid Sha Baolao culture medium of sterilizing, by the candida utili bacterium of 10% inoculum concentration access activation, temperature 30 DEG C, rotating speed 180r/min, pressure 0.05MPa, cultivate 24h ~ 36h, liquid candida utili bacteria preparation can be obtained.
Consisting of of described solid Sha Baolao medium nutrient content: peptone 10 parts, glucose 40 parts, 20 parts, agar, distilled water 1000 parts, pH6.8.Sterilising temp 115 DEG C, sterilization time 20min.
Consisting of of described liquid Sha Baolao medium nutrient content: peptone 10 parts, glucose 40 parts, distilled water 1000 parts, pH6.8.Sterilising temp 115 DEG C, sterilization time 20min.
Mixed with water by wheat bran, the two mass ratio is 1:0.5, and high-temperature sterilization cools, and obtains sterile carrier inserts.
By liquid candida utili bacteria preparation and sterile carrier inserts Homogeneous phase mixing, the two weight ratio is 1:10, and 30 DEG C ~ 37 DEG C fermentation 24h ~ 36h post-dryings are pulverized, and can obtain candida utili bacterium bacterium powder.
Second step: adding of obtained each zymocyte liquid and additive carrier inserts.By wheat bran with water in mass ratio for 1:0.5 mixes, high-temperature sterilization cool, obtain sterile carrier inserts.By each zymocyte liquid respectively with sterile carrier inserts in mass ratio for 1:10 Homogeneous phase mixing ferments.
3rd step: drying, pulverizing, mixing (bacillus subtilis bacterium powder, bafillus natto bacterium powder, Lactobacillus casei bacterium powder, candida utili bacterium bacterium powder are mixed in the ratio of 0.5:0.5:1:1), obtained composite probiotics preparations powder-type.
4th step: pack is preserved.
The application of embodiment 2 Shelled Turtle Trionyx Sinensis composite probiotics feed additive
Test site is in the Yueyang Linxiang cultivation base of Hunan Huang Xin Aqua Sciences Inc., and subjects is Shelled Turtle Trionyx Sinensis, applies product of the present invention and to feed Shelled Turtle Trionyx Sinensis, on its growth and benefit to affect comparative analysis as follows:
This test is based on commercial feed formula.In basal feed, add composite probiotics preparations, compound formulation pulverized 60 orders.Additive arranges 4 gradients, and (0mg/kg, 500mg/kg, 1000mg/kg, 2000mg/kg ﹚, test is divided into 4 groups, often organizes 3 repetitions, totally 12 groups, and often organizing pond specification is 4m × 10m × 1.5m, puts 1200 in each pond in a suitable place to breed.Throw something and feed every day 3 times (7:00-8:00,11:00-12:00,18:00-19:00), free choice feeding.Test 20 weeks by a definite date.Clearing rate of body weight gain, specific growth rate, feed coefficient, survival rate, income and net income, and be recorded in detail.
Rate of body weight gain (%)=100 × (whole opisthosoma weighs an original body mass)/original body mass;
Specific growth rate (%/d)=100 × (㏑ whole opisthosoma Chong mono-㏑ original body mass)/test number of days.
Total augment weight amount (g) of total feed consumption rate (g) in feed coefficient=experimental period/in experimental period
Survival rate (%)=survive and count/put sum × 100% in a suitable place to breed
Income=output × unit price
Net income=income-fry cost-manually wait cost
The results are shown in Table 1.
Table 1 Shelled Turtle Trionyx Sinensis growth performance contrasts
As can be seen from Table 1, the opisthosoma of interpolation compound probiotic process 2, process 3 is heavy, rate of body weight gain, specific growth rate are all significantly higher than control group and process 1(P < 0.01), feed coefficient is significantly lower than control group and process 1(P < 0.01), the rate of body weight gain wherein processing 2 wants high by 29.25% compared with control group; Survival rate all the other each group is all significantly higher than control group (P < 0.05), wherein processes 2 and reaches maximum (P < 0.01).
Soft-shelled turtle seedling price 12 yuan/kg then, feed 13 yuan/kg, employee takes 1000 yuan/hm
2, put fishing expenses of labour 600 yuan, probio expense 300 yuan, pool hire charge 200 yuan, various incidentals 5000 yuan; Shelled Turtle Trionyx Sinensis commercial value 40 yuan/kg.To the Economic and Efficiency Analysis of each processed group, wherein the economic benefit of process 2 is the highest, and net income is 295.54 ten thousand yuan/hm
-2, input and output ratio is 1: 1.86.Concrete benefit details is in table 2.
The culture benefit of table 2 Different adding amount
Claims (3)
1. Shelled Turtle Trionyx Sinensis probiotic feed additive, it is characterized in that: described probiotic feed additive is made up of the bacterium powder of bacillus subtilis, bafillus natto, Lactobacillus casei, candida utili bacterium, the bacterium powder of bacillus subtilis, bafillus natto, Lactobacillus casei, candida utili bacterium is obtained using the wheat bran of sterilizing as absorption carrier respectively, and its constituent mass ratio is: 0.4 ~ 0.6:0.4 ~ 0.6:0.8 ~ 1.2:0.8 ~ 1.2; The viable count of bacterium powder is more than or equal to: bacillus subtilis 1 × 10
8cFU/g, bafillus natto 1 × 10
9cFU/g, Lactobacillus casei 1 × 10
11cFU/g, candida utili bacterium 1 × 10
8cFU/g; Preparation process is as follows:
(1) bacillus subtilis, numbering: CICC 21746,
bacilluss
ubtilisthe preparation of preparation
The solid ordinary culture medium activation of the bacillus subtilis sterilizing of first inclined-plane being preserved, through 30 DEG C ~ 37 DEG C activation culture 24h ~ 36h, then on the liquid ordinary culture medium of sterilizing, by the bacillus subtilis of 10% inoculum concentration access activation, temperature 30 DEG C, rotating speed 180r/min, pressure 0.05MPa, cultivate 24h ~ 36h, liquid bacillus subtilis formulation can be obtained;
Consisting of of described solid ordinary culture medium nutritional labeling: 5 parts, sodium chloride, beef extract 3 parts, peptone 10 parts, 20 parts, agar, distilled water 1000 parts, PH 7.2 ~ 7.4; Sterilising temp 115 DEG C, sterilization time 20min;
Consisting of of described liquid ordinary culture medium nutritional labeling: 5 parts, sodium chloride, beef extract 3 parts, peptone 10 parts, distilled water 1000 parts, PH7.2 ~ 7.4; Sterilising temp 115 DEG C, sterilization time 20min;
Mixed with water by wheat bran, the two mass ratio is 1:0.5, and high-temperature sterilization cools, and obtains sterile carrier inserts;
By liquid bacillus subtilis formulation and sterile carrier inserts Homogeneous phase mixing, the two weight ratio is 1:10, and 30 DEG C ~ 37 DEG C ventilating fermentation 24h ~ 36h post-dryings are pulverized, and can obtain bacillus subtilis bacterium powder; Viable count is more than or equal to 1 × 10
8cFU/g;
(2) bafillus natto, numbering: CICC 20443,
bacillus nattothe preparation of preparation
The solid Sha Baolao culture medium activation of the bafillus natto sterilizing of first inclined-plane being preserved, through 30 DEG C ~ 37 DEG C activation culture 24h ~ 36h, then on the liquid Sha Baolao culture medium of sterilizing, by the bafillus natto of 10% inoculum concentration access activation, temperature 37 DEG C, rotating speed 180r/min, pressure 0.05MPa, cultivate 24h ~ 36h, liquid natto bacillus preparation can be obtained;
Consisting of of described solid Sha Baolao medium nutrient content: peptone 10 parts, maltose 40 parts, 20 parts, agar, distilled water 1000 parts, PH6.8, sterilising temp 115 DEG C, sterilization time 20min;
Consisting of of described liquid Sha Baolao medium nutrient content: peptone 10 parts, maltose 40 parts, distilled water 1000 parts, PH 6.8; Sterilising temp 115 DEG C, sterilization time 20min;
Mixed with water by wheat bran, the two mass ratio is 1:0.5, and high-temperature sterilization cools, and obtains sterile carrier inserts;
By liquid natto bacillus preparation and sterile carrier inserts Homogeneous phase mixing, the two weight ratio is 1:10, and 30 DEG C ~ 37 DEG C ventilating fermentation 24h ~ 36h post-dryings are pulverized, and can obtain bafillus natto bacterium powder; Viable count is more than or equal to 1 × 10
9cFU/g;
(3) Lactobacillus casei, numbering: CICC 6105,
lactobacillus caseithe preparation of preparation
The solid MRS culture medium activation of the Lactobacillus casei sterilizing of first inclined-plane being preserved, through 30 DEG C ~ 37 DEG C activation culture 24h ~ 36h, then on the liquid MRS culture medium of sterilizing, by the Lactobacillus casei of 10% inoculum concentration access activation, temperature 37 DEG C, quiescent culture 24h ~ 36h, can obtain liquid Lactobacillus casei preparation;
Consisting of of described solid MRS medium nutrient content: peptone 10 parts, beef extract 10 parts, yeast extract 5 parts, diammonium hydrogen citrate 2 parts, glucose 20 parts, Tween 80 1 part, sodium acetate 5 parts, dipotassium hydrogen phosphate 2 parts, 0.58 part, magnesium sulfate, manganese sulfate 0.25 part, 18 parts, agar, distilled water 1 000 parts, PH:6.2 ~ 6.6; Sterilising temp 121 DEG C, sterilization time 15min;
Consisting of of described liquid MRS medium nutrient content: peptone 10 parts, beef extract 10 parts, yeast extract 5 parts, diammonium hydrogen citrate 2 parts, glucose 20 parts, Tween 80 1 part, sodium acetate 5 parts, dipotassium hydrogen phosphate 2 parts, 0.58 part, magnesium sulfate, manganese sulfate 0.25 part, distilled water 1 000 parts, PH6.2 ~ 6.6, sterilising temp 121 DEG C, sterilization time 15min;
Mixed with water by wheat bran, the two mass ratio is 1:0.5, and high-temperature sterilization cools, and obtains sterile carrier inserts;
By liquid Lactobacillus casei preparation and sterile carrier inserts Homogeneous phase mixing, the two weight ratio is 1:10, and 30 DEG C ~ 37 DEG C sealing and fermenting 24h ~ 36h post-dryings are pulverized, and can obtain Lactobacillus casei bacterium powder; Viable count is more than or equal to 1 × 10
11cFU/g;
(4) candida utili bacterium numbering: CICC 1314,
candida utilisthe preparation of preparation
The solid Sha Baolao culture medium activation of the candida utili bacterium sterilizing of first inclined-plane being preserved, through 28 DEG C ~ 30 DEG C activation culture 24h ~ 36h, then on the liquid Sha Baolao culture medium of sterilizing, by the candida utili bacterium of 10% inoculum concentration access activation, temperature 30 DEG C, rotating speed 180r/min, pressure 0.05MPa, cultivate 24h ~ 36h, liquid candida utili bacteria preparation can be obtained;
Consisting of of described solid Sha Baolao medium nutrient content: peptone 10 parts, glucose 40 parts, 20 parts, agar, distilled water 1000 parts, PH6.8; Sterilising temp 115 DEG C, sterilization time 20min;
Consisting of of described liquid Sha Baolao medium nutrient content: peptone 10 parts, glucose 40 parts, distilled water 1000 parts, PH6.8; Sterilising temp 115 DEG C, sterilization time 20min;
Mixed with water by wheat bran, the two mass ratio is 1:0.5, and high-temperature sterilization cools, and obtains sterile carrier inserts;
By liquid candida utili bacteria preparation and sterile carrier inserts Homogeneous phase mixing, the two weight ratio is 1:10, and 30 DEG C ~ 37 DEG C fermentation 24h ~ 36h post-dryings are pulverized, and can obtain candida utili bacterium bacterium powder; Candida utili bacterium viable count is more than or equal to 1 × 10
8cFU/g;
(5) preparation of composite probiotics preparations
By above-mentioned bacillus subtilis bacterium powder, bafillus natto bacterium powder, Lactobacillus casei bacterium powder, candida utili bacterium bacterium powder Homogeneous phase mixing in proportion, obtain composite probiotics preparations powder form.
2. Shelled Turtle Trionyx Sinensis probiotic feed additive according to claim 1, is characterized in that, described composite probiotics preparations adds in Shelled Turtle Trionyx Sinensis feed by the dosage of 1000mg/kg and uses.
3. Shelled Turtle Trionyx Sinensis probiotic feed additive according to claim 1, is characterized in that all completing with stainless steel fermentation tank in fermentation production process.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410039520.3A CN103783321B (en) | 2014-01-27 | 2014-01-27 | Shelled Turtle Trionyx Sinensis probiotic feed additive and preparation method |
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| CN105265842A (en) * | 2015-10-23 | 2016-01-27 | 樊德喜 | Chinese herbal medicine containing feed for turtles and preparation method of feed |
| CN106361905B (en) * | 2016-08-30 | 2019-07-05 | 福建贝迪药业有限公司 | A kind of livestock and poultry Fuzheng Jiedu compound Chinese herb probiotics and preparation method thereof |
| CN107568159A (en) * | 2017-10-16 | 2018-01-12 | 孝感市鲁建农业开发有限公司 | A kind of cultural method for promoting Shelled Turtle Trionyx Sinensis growth |
| CN107736479A (en) * | 2017-11-06 | 2018-02-27 | 潍坊中创生物科技有限公司 | A kind of taurine product and preparation method thereof |
| CN108013248A (en) * | 2017-12-28 | 2018-05-11 | 广东省农业科学院动物科学研究所 | One boar fermentable fiber feed and preparation method thereof |
| CN109156624A (en) * | 2018-09-28 | 2019-01-08 | 河南科技大学 | A kind of immunopotentiator and the preparation method and application thereof improving soft-shelled turtle premunition |
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| CN1568747A (en) * | 2004-04-27 | 2005-01-26 | 广州市希普生物饲料有限公司 | Direct feeding viable bacteria preparation and method for preparing it |
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| CN102948621A (en) * | 2012-11-12 | 2013-03-06 | 东北农业大学 | Prebiotic peptide biological feed additive and preparation method and application thereof |
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| CN1800348A (en) * | 2005-12-14 | 2006-07-12 | 山东大学 | Multifunctional probiotic preparation and its preparation method |
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