Background technology
Porcine epizootic diarrhea and transmissible gastroenteritis of swine are worldwide widely current, especially in Korea S, and the Asian countries such as Japan and China.It has carried out economic loss seriously to industrial belt of raising pigs.PED causes that the survival rate of pig reduces, regularity is poor, slows down the speed of growth, weightening finish reduces, postpone Time To Market, reduce efficiency of feed utilization, increase and additionally consume, reduce domestic animal premunition, increase susceptibility to antibacterial, virus, increase sickness rate, improves medical expense, domestic animal dehydration, shock, dead, pig have loose bowels the serious consequences such as weight loss.
At present, for porcine epizootic diarrhea and transmissible gastroenteritis of swine also without definite effective Therapeutic Method, therefore to the prevention and control of these two kinds of diseases, the policy adopting " put prevention first, support anti-combination " more.In order to effectively control porcine epizootic diarrhea and transmissible gastroenteritis of swine, China has developed the vaccine for preventing and treating porcine epizootic diarrhea and transmissible gastroenteritis of swine, but to its diluent, particularly be applicable to transmissible gastroenteritis of swine simultaneously, the research of epidemic diarrhea bigeminal live vaccine diluent is less, currently available vaccines, existing vaccines diluent effect is not good for the effect of transmissible gastroenteritis of swine, epidemic diarrhea bigeminal live vaccine yet.
Summary of the invention
For above-mentioned technical problem, the invention provides a kind of transmissible gastroenteritis of swine, epidemic diarrhea bigeminal live vaccine diluent, can immune effect of vaccine be improved, make swinery obtain better immune protective efficiency.Concrete technical scheme is:
Transmissible gastroenteritis of swine, epidemic diarrhea live vaccine diluent, prepare gained by following steps:
(1) A liquid preparation
Containing the Na of KCl, 3.58g of NaCl, 0.2g of 8g in often liter of A liquid
2hPO
412H
2the KH of O, 0.24g
2pO
4, above substance dissolves, in water for injection, carries out sterilization treatment under 121 DEG C of temperature, 100 kPas of pressure conditions;
(2) B liquid preparation
Aqueous phase: the Na of KCl, the 3.58g of NaCl, 0.2g containing 8g in every premium on currency
2hPO
412H
2the KH of O, 0.24g
2pO
4, above substance dissolves is in water for injection; Add astragalus polysaccharides, propolis, Tween 80 again, adding rear astragalus polysaccharides concentration is 0.5 ~ 5%w/v, and propolis concentration is 10 ~ 100mg/ml, and Tween 80 concentration is 1 ~ 5%w/v, for subsequent use after filtration sterilization;
Oil phase: for subsequent use after 121 DEG C, mineral oil, 100 kPas of pressure sterilizings;
Oil phase slowly to be added in aqueous phase and to stir, oil phase and aqueous phase weight ratio 1: 2, re-using high-shear emulsion machine after oil phase adds and obtain B liquid with 4000r/min emulsifying 30min;
(3) use of diluent: during use, A liquid and B liquid to be mixed homogeneously in mass ratio at 1: 1.
Transmissible gastroenteritis of swine provided by the invention, epidemic diarrhea bigeminal live vaccine diluent, can improve immune effect of vaccine, makes swinery obtain better immune protective efficiency.
Detailed description of the invention
The specific embodiment of the present invention is described in conjunction with the embodiments.
Transmissible gastroenteritis of swine, epidemic diarrhea live vaccine diluent, prepare gained by following steps:
(1) A liquid preparation
Containing the Na of KCl, 3.58g of NaCl, 0.2g of 8g in often liter of A liquid
2hPO
412H
2the KH of O, 0.24g
2pO
4, above substance dissolves, in water for injection, carries out sterilization treatment under 121 DEG C of temperature, 100 kPas of pressure conditions;
(2) B liquid preparation
Aqueous phase: the Na of KCl, the 3.58g of NaCl, 0.2g containing 8g in every premium on currency
2hPO
412H
2the KH of O, 0.24g
2pO
4, above substance dissolves is in water for injection; Add astragalus polysaccharides, propolis, Tween 80 again, adding rear astragalus polysaccharides concentration is 0.5 ~ 5%w/v, and propolis concentration is 10 ~ 100mg/ml, and Tween 80 concentration is 1 ~ 5%w/v, for subsequent use after filtration sterilization;
Oil phase: for subsequent use after 121 DEG C, mineral oil, 100 kPas of pressure sterilizings;
Oil phase slowly to be added in aqueous phase and to stir, oil phase and aqueous phase weight ratio 1: 2, re-using high-shear emulsion machine after oil phase adds and obtain B liquid with 4000r/min emulsifying 30min;
(3) use of diluent: during use, A liquid and B liquid to be mixed homogeneously in mass ratio at 1: 1.
The diluent of above embodiment gained and comparison example are carried out contrast experiment:
1, vaccine dilution: transmissible gastroenteritis, epidemic diarrhea bigeminal live vaccine are diluted to 1 part/ml by the diluent using the present embodiment to obtain, phosphate buffer PBS respectively.
2, immunity contrast
(1) 1 ~ 3 age in days piglet: 1 ~ 3 age in days piglet 15, is divided into A1, B1, C13 group, often organizes 5.
Vaccine after the diluted that the existing the present embodiment of A1 group injection obtains, 1.0ml/ head; Vaccine after B1 group injection PBS dilution, 1.0ml/ head; PBS is as blank group for the injection of C1 group, 1.0ml/ head.0th, 7,14,21,28 and 35 day separation of serum of taking a blood sample respectively after immunity, measures PED and TGE NAT, calculates the geometrical mean of often group piglet antibody titer and carries out comparative analysis.
(2) sow: antenatal 28 ~ 35 age in days sow 15, is divided into A2, B2, C23 group, often organizes 5.Vaccine after the diluted that A2 group group injection embodiment obtains; Vaccine after B2 group injection PBS dilution, 1.0ml/ head; PBS is as blank group for the injection of C2 group, 1.0ml/ head.After immunity, the 0th, 14,28,35 take a blood sample separation of serum respectively, measure PED and TGE NAT, calculate the geometrical mean of often organizing sow antibody titer and carry out comparative analysis.
3, interpretation of result
(1) 1 ~ 3 age in days piglet: diluent group 1 ~ 3 age in days TGE antibody horizontal that the 7th, 14,21,28 and 35 day produces after immunity is apparently higher than PBS group and blank group; The PED antibody horizontal that after immunity, the 7th, 14,21,28 and 35 day produces is apparently higher than PBS group and blank group, and the 7th day diluent group PED antibody titer geometrical mean, higher than 1: 32, the results are shown in Figure 1 and Fig. 2 after vaccine immunity.
(2) sow: after the immunity of diluent group sow the 14th, 28,35 and produce the same day TGE antibody horizontal apparently higher than PBS group and blank group; After immunity, the 14th, 28 and 35 day PED antibody horizontal is apparently higher than PBS group and matched group, the results are shown in Figure 3 and Fig. 4.Sow PED and TGE High antibody level, contribute to primiparity piglet and obtain protection by maternal antibody, thus reduce or to alleviate the disease caused by transmissible gastroenteritis or epidemic diarrhea even dead.