CN1343482A - Calmette's vaccine injection method for enhancing vaccine immunity of animals - Google Patents
Calmette's vaccine injection method for enhancing vaccine immunity of animals Download PDFInfo
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Abstract
An injection method of TB vaccine to strengthen the animal vaccine immunization injects the TB vaccine to the animals to be immuned before or at the same time of the vaccine immunization injection. To use TB vaccine in stead of traditional medicine immunization can effectively strengthen the immunization ability of vaccine injection. The method is suitable for the animal vaccine immunization injection especially for the pig and chicken.
Description
The present invention relates to the bacillus calmette-guerin vaccine injecting method that a kind of immune effect of vaccine to animal (poultry, fowl etc.) plays potentiation.
In order to strengthen vaccine immunity of animals injection effect, people taked the way of medicine cooperation more, for example: CN1200272 immunostimulant that can suppress tumor growth in the past; CN1176139 chicken Marek's disease vaccine immunopotentiator and preparation technology; The preparation method of CN1091639 rabies vaccine specific immunity enhancing agent.These methods are because temporary transient or long-term in animal body the accumulation of the effect of paying of medicine and some composition has certain defective.Do not find to adopt the report of bacillus calmette-guerin vaccine at present as the application of vaccine immunopotentiator in animal.
The purpose of this invention is to provide a kind of injecting method that adopts bacillus calmette-guerin vaccine as the bacillus calmette-guerin vaccine of animal vaccine immunopotentiator.
The present invention is achieved in that and a kind of vaccine immunity of animals is played the bacillus calmette-guerin vaccine injecting method of potentiation, it is characterized in that: bacillus calmette-guerin vaccine is injected on one's body the animal for the treatment of immunity inoculation before vaccine injection or with vaccine simultaneously.Bacillus calmette-guerin vaccine (BCG) is a kind of broad-spectrum immunocyte activator, can stimulate the pluripotent stem cell of bone marrow, make it to develop into immunologically competent cell, promote the differentiation and proliferation of T, B, K, NK cell and macrophage, thereby improve the immunity of body significantly with immunologic function.The nonspecific immunostimulant of this broad-spectrum has general significance aspect the enhancing of animal immune ability.Yet the protection effect of non-specific immunity is limited, and therefore, it is unsatisfactory only to adopt bacillus calmette-guerin vaccine to control the disease effects of animal; Have only the nonspecific immunity factor of bacillus calmette-guerin vaccine and specific immunity vaccine combined and carry out immunity and just can reach good result.That is to say, bacillus calmette-guerin vaccine could be improved the immune effect of this kind animal effectively as the reinforcing agent of special vaccine immunity.In addition, find bacillus calmette-guerin vaccine dosage size after deliberation or reuse number of times not to be directly proportional with immune effect, strong excessively stimulation might be brought out immunosuppressant on the contrary.Therefore, having only specific immunity factor with the specific immune vaccine of the specified disease of the nonspecific immunity factor of bacillus calmette-guerin vaccine and animal to combine the immunity of carrying out animal handles and just can receive significant immune effect.At animal vaccine (for example: (about 1-10 days) injection of BCG vaccine in advance before the injection chicken coccidiosis vaccine), activate the non-specific factor of animal immune system earlier, for the activation of the specific factors of this system (for example: the specific immunity effect of chicken coccidiosis vaccine) lay a solid foundation, thereby (for example: young chicken) intravital immune system obtains not only generally but also targetedly activating, significantly the enhance immunity effect to make animal.Certainly, (for example: injection simultaneously swine Fever Vaccine) (comprising: injection simultaneously with bacillus calmette-guerin vaccine and animal vaccine, it is also good successively to inject effect successively, this be since in the intravital immune system of animal nonspecific immunity factor and specific immunity factor be activated the cause that plays a role simultaneously simultaneously.But, if injection of BCG vaccine again when treating that animal specificity vaccine injection effect begins to descend, then because in the bacillus calmette-guerin vaccine effect performance, the intravital specific immunity effect of animal weakens, even disappear, processing does not just reach and forms nonspecific immunity and the complementary effect of specific immunity in animal body like this.
The present invention can be specific like this: the inoculation of animal is generally carried out in the childhood of animal.For the young pig in 20～40 day age, the injection volume of bacillus calmette-guerin vaccine be 0.1～0.2 milligram/more suitable; As the immunostimulant of swine Fever Vaccine, the injection of bacillus calmette-guerin vaccine and swine Fever Vaccine inoculation are carried out simultaneously.For the young chicken in 7～14 day age, the injection volume of bacillus calmette-guerin vaccine be 0.025～0.05 milligram/only more suitable; As the effect enhancer of chicken coccidiosis vaccine, bacillus calmette-guerin vaccine can be injected simultaneously with the chicken coccidiosis vaccine or carry out in seven days in advance.
The present invention injects before vaccination or simultaneously because bacillus calmette-guerin vaccine is replaced medicine as animal vaccine immunopotentiator, has improved the immune effect of vaccination effectively; (for example: the coccidiosis of chicken) effect is particularly good for the animal epidemic of especially not too strong to some immune effect or poor stability.
Embodiment 1: bacillus calmette-guerin vaccine is as the immunostimulant injection of swine Fever Vaccine.
Test pig: the healthy Du-length of 40 ages in days-Dasanyuan hybridization ablactation commodity piglet, all about 10kg of body weight, male or female, the feeds utilized piglet that produces for commercial feed factory cooperates complete feed.
Hog cholera lapinised virus vaccine: available from Guangdong Province's biologics factory, lot number 9942 with the dissolving of 12.5mL sterile saline, uses up in 30 minutes.
Immunostimulant: the calf thymus element, Wuhan Mei Shan biochemical product factory produces, and lot number 980401, content are 2mg/mL; Lyophilized bacillus calmette-guerin vaccine, the institute of biological products, Guangdong Province produces, lot number 981102, content is 75mg/mL, faces with preceding to dissolve with the 10mL sterilized water for injection by every of explanation, uses up in 30 minutes.1, test grouping and processing:
Test grouping and processing see Table 1.This test divides thymosin group (T), bacillus calmette-guerin vaccine group (B), thymosin+swine Fever Vaccine group (TC), bacillus calmette-guerin vaccine+swine Fever Vaccine group (BC), swine Fever Vaccine group (C) and normal saline (N), the test pig number, immunizing dose, immune time and vaccination ways are by arranging to carry out in the table.Table 1 test grouping and processing
Annotate: T refers to thymosin, and B refers to bacillus calmette-guerin vaccine, and TC refers to thymosin+swine Fever Vaccine, and BC refers to bacillus calmette-guerin vaccine+swine Fever Vaccine, and C refers to swine Fever Vaccine, and N pointed injection normal saline 2, test detect index
| Handle | ??T | ??B | ?TC | ?BC | ?C | ?N |
| Test pig number (head) | ??3 | ??3 | ??3 | ??3 | ??3 | ??3 |
| Immunizing dose | ??2mg | ??7.5mg | 2mg+4 head part | 7.5mg+4 head part | 4 parts | ??1mL |
| Immune time | ??1 | ??1 | ??1 | ??1 | ??1 | ??1 |
| Vaccination ways | Mix the back subcutaneous injection | Mix the back subcutaneous injection | Mix the back subcutaneous injection | Mix the back subcutaneous injection | Mix the back subcutaneous injection | Mix the back subcutaneous injection |
Blood sampling is at the date: 0 (before the immunity), 3,7,14,21,28 (immunity backs) day,, institute's blood-sample withdrawal detects index to be had: (1) pig PBNC level of conversion; (2) numeration of leukocyte; (3) active T lymphocyte test (4) serum IgG, IgM level.(5) serum bactericidal test; (6) the mutual immunoprotection test of rabbit body; Wherein, (1), (2), (3) project detect 0,3,7,10 day serum, (4) detection 0,3,7,10,14, the IgM content in sky and 0,3,7,10,14,21,28 day IgG content, and (5) (6) project detects the 7th day serum.3, result of the test: 1. immunostimulant is to the influence of pig PBMC breeder reaction
The results are shown in Table 2, along with the variation difference of age in days is not remarkable.All the other each groups are the most obvious with BC, TC, B group ascendant trend.Immunity back the 7th day, BC, TC, B group lymphocyte transformation level reach peak, than C group significant difference (P＜0.01).T group and B group are after immunity, the lymphocyte transformation level is in rising trend, and organize than difference remarkable (P＜0.05) with N, B and BC group difference are not remarkable, T and TC group is at immunity significant difference that influences to pig PBMC breeder reaction after 7 days, each is organized the lymphocyte transformation level and all is in various degree decline trend after 7 days in immunity, to back 10 days of immunity, and lymphocyte transformation level difference not significantly (P＞0.05) between each group.Table 2 immunostimulant is to the influence of pig PBMC breeder reaction
Annotate: (1) T: calf thymus element; B: bacillus calmette-guerin vaccine; TC: calf thymus element+swine Fever Vaccine; BC: bacillus calmette-guerin vaccine+swine Fever Vaccine; C: hog cholera immune contrast; N: blank normal healthy controls.(2) numeral be x ± SE in the table, and the new multipole difference of The data Deng Kenshi method of inspection (DMRF method) carries out multiple comparisons, and letter is statistical result in the round bracket, and the alphabetical person's significant difference inequality (a=0.05) of same column is identical or do not have the alphabetic flag person's difference is not remarkable.2. immunostimulant is to the influence of total white blood cells
| Blood sampling time | |||||
| Handle | Before the injection | After the injection | |||
| 0 day | 3 days | 7 days | 10 days | 14 days | |
| ??T | ?0.242±0.018 | ?0.451±0.031 ????(a) | 0.376±0.024 ????(c) | 0.170±0.061 | ??0.099±0.002 |
| ??B | ?0.242±0.018 | ?0.413±0.038 ????(a) | 0.710±0.379 ????(b) | 0.197±0.045 | ?0.091±0.026 |
| ?TC | ?0.242±0.018 | ?0.439±0.034 ????(a) | 0.691±0.012 ????(b) | 0.174±0.042 | ?0.109±0.005 |
| ?BC | ?0.242±0.018 | ?0.451±0.061 ????(a) | 0.892±0.193 ????(a) | 0.163±0.026 | ??0.086±0.035 |
| ??C | ?0.242±0.018 | ?0.432±0.020 ????(a) | 0.350±0.021 ????(c) | 0.160±0.087 | ?0.099±0.029 |
| ?N | ?0.242±0.018 | ?0.224±0.004 ????(b) | 0.150±0.028 ????(d) | 0.219±0.033 | ?0.117±0.021 |
The results are shown in Table 3.The total white blood cells of normal healthy controls changes not quite in the detection period, can find out that from leukocyte change curve chart bacillus calmette-guerin vaccine, thymosin have certain stimulation to leukocytic generation, and the influence in 3-7 days of immunity back is (P＜0.05) significantly.Influence not remarkable (P＜0.05) after the 10th day.Table 3 immunostimulant is to the influence of total white blood cells
Annotate: (1) TC: calf thymus element+swine Fever Vaccine; BC: bacillus calmette-guerin vaccine+swine Fever Vaccine; T: calf thymus element; B: bacillus calmette-guerin vaccine; C: hog cholera immune contrast; N: blank normal healthy controls.(2) numeral be x ± SE in the table, and the new multipole difference of The data Deng Kenshi method of inspection (DMRF method) carries out multiple comparisons, and letter is statistical result in the round bracket, and the alphabetical person's significant difference inequality (a=0.05) of same column is identical or do not have the alphabetic flag person's difference is not remarkable.3. immunostimulant is to the lymphocytic influence of active T
| Blood sampling time | |||||
| Handle | Before the injection | After the injection | |||
| 0 day | 3 days | 7 days | 10 days | 14 days | |
| ????TC | ??331±17 | ??377±15 ????(a) | ????354±6 ????(a) | ??323±29 | ??333±23 |
| ????BC | ??331±17 | ??390±15 ????(a) | ??375±58 ????(a) | ??343±37 | ??306±41 |
| ????T | ??331±17 | ??364±8 ????(b) | ??336±16 | ??323±17 | ??328±5 |
| ????B | ??331±17 | ??379±31 ????(a) | ??348±22 | ??333±11 | ??331±38 |
| ????C | ??331±17 | ??354±26 ????(b) | ??364±52 ????(a) | ??353±12 | ??341±30 |
| ????N | ??331±17 | ??343±15 ????(B) | ??336±20 ????(b) | ??338±23 | ??339±15 |
The results are shown in Table 4, the E-rosette forming rate of normal healthy controls is the level curve fluctuation in detection period, the E-rosette forming rate of each immune group all is increasing progressively and the 3rd day left and right sides peaking after immunity in various degree, wherein, be remarkable difference between BC, TC and T, B, C, do not have significant difference between BC and TC.The result shows, bacillus calmette-guerin vaccine and thymosin all can the utmost point significantly (P＜0.01) improve and prolong the E-rosette forming rate of immune swine.Table 4 immunostimulant is to the lymphocytic influence of active T
Annotate: (1) T: calf thymus element; B: bacillus calmette-guerin vaccine; TC: calf thymus element+swine Fever Vaccine; BC: bacillus calmette-guerin vaccine+swine Fever Vaccine; C: hog cholera immune contrast; N: blank normal healthy controls.(2) numeral be x ± SE in the table, and the new multipole difference of The data Deng Kenshi method of inspection (DMRF method) carries out multiple comparisons, and letter is statistical result in the round bracket, and the alphabetical person's significant difference inequality (a=0.05) of same column is identical or do not have the alphabetic flag person's difference is not remarkable.4. immunostimulant is to the influence of serum IgM
| Blood sampling time | |||||
| Handle | Before the injection | After the injection | |||
| 0 day | 3 days | 7 days | 10 days | 14 days | |
| ??T | ??2.03±0.54 | ??3.50±0.87 ????(c) | ??3.33±1.15 ????(a) | ??1.17±0.29 ????(b) | ??2.50±1.32 ????(a) |
| ??B | ??2.03±0.54 | ??2.67±0.29 ????(c) | ??2.00±0.00 ????(b) | ??1.00±0.05 ????(b) | ??1.33±0.29 ????(b) |
| ??TC | ??2.03±0.54 | ??6.00±1.32 ????(a) | ??4.00±1.50 ????(a) | ??1.17±0.29 ????(b) | ??1.67±0.58 ????(b) |
| ??BC | ??2.03±0.54 | ??6.33±1.44 ????(a) | ??3.50±0.48 ????(a) | ??2.50±1.19 ????(a) | ??1.83±1.03 ????(b) |
| ??C | ??2.03±0.54 | ??5.00±2.29 ????(b) | ??4.33±0.76 ????(a) | ??1.17±0.29 ????(b) | ??1.33±0.58 ????(b) |
| ??N | ??2.03±0.54 | ??1.75±0.35 ????(d) | ??2.67±0.29 ????(b) | ??0.83±0.29 ????(b) | ??2.00±0.50 ????(b) |
The results are shown in Table 5.Because of the pig of 40 ages in days still has certain maternal antibody, so in back 3 days of the immunity, except that N, T, B group, serum IgM content of all the other each groups all have the rapid reduction with degree, this shows that the part maternal antibody is neutralized by swine Fever Vaccine virus.After the immunity 3 days, immunity is respectively organized serum IgM content and is begun rapid rising, and the TC group rises to the peak with the BC group, is remarkable difference with the C group; The serum content of C group rises relatively slow, and immunity back the 10th genius peaking descend rapidly subsequently, and the BC group is kept higher serum IgM antibody level always, to the 14th day, still is in the positive value scope.Table 5 immunostimulant is to the influence of serum IgM antibody
Annotate: (1) T: calf thymus element; B: bacillus calmette-guerin vaccine; TC: calf thymus element+swine Fever Vaccine; BC: bacillus calmette-guerin vaccine+swine Fever Vaccine; C: hog cholera immune contrast; N: blank normal healthy controls.(2) numeral be x ± SE in the table, and the new multipole difference of The data Deng Kenshi method of inspection (DMRF method) carries out multiple comparisons, and letter is statistical result in the round bracket, and the alphabetical person's significant difference inequality (a=0.05) of same column is identical or do not have the alphabetic flag person's difference is not remarkable.5. immunostimulant is to the influence of serum IgG content
| Blood sampling time (n=54) | |||||
| Handle | Before the injection | After the injection | |||
| 0 day | 3 days | 7 days | 10 days | 14 days | |
| ??T | ?0.216±0.027 | ?0.204±0.015 ????(a) | 0.188±0.008 ????(c) | 0.141±0.007 ????(c) | 0.073±0.005 ????(c) |
| ??B | ?0.216±0.027 | ?0.218±0.005 ????(a) | 0.173±0.010 ????(c) | 0.150±0.023 ????(c) | 0.077±0.106 ????(c) |
| ??TC | ??0.216±0.027 | ??0.034±0.020 ????(b) | 0.272±0.064 ????(b) | 0.194±0.021 ????(b) | 0.130±0.033 ????(b) |
| ??BC | ??0.216±0.027 | ??0.044±0.035 ????(b) | 0.344±0.034 ????(a) | 0.330±0.046 ????(a) | 0.220±0.025 ????(a) |
| ??C | ?0.216±0.027 | ?0.064±0.010 ????(b) | 0.137±0.010 ????(c) | 0.237±0.107 ????(b) | 0.083±0.061 ????(c) |
| ??N | ?0.216±0.027 | ?0.194±0.014 ????(a) | 0.161±0.028 ????(c) | 0.125±0.047 ????(c) | 0.059±0.024 ????(c) |
The results are shown in Table 6.T, B, N organize in detection period, and serum IgG OD value is downward trend gradually.In the time of back 3 days, the serum IgG antibody level drops to minimum on duty three immune group (TC, BC, C group) in immunity, and 3-14 days thereafter, antibody rose gradually, to back about 14 days of immunity, reached peak, and continually and steadily to 28 talent's declines.Wherein the BC group is higher than positive initial value at back 7 days serum IgG contents of immunity, is significant difference (P＜0.05) with the B group, but is the significant difference (P＜0.05) in back 14 days except that immunity between TC group and C group, and serum IgG content difference is not remarkable between all the other ages in days.Table 6 immunostimulant is to the result that influences of serum IgG content
Annotate: (1) T: calf thymus element; B: bacillus calmette-guerin vaccine; TC: calf thymus element+swine Fever Vaccine; BC: bacillus calmette-guerin vaccine+swine Fever Vaccine; C: hog cholera immune contrast; N: blank normal healthy controls.(2) numeral be x ± SE in the table, and the new multipole difference of The data Deng Kenshi method of inspection (DMRF method) carries out multiple comparisons, and letter is statistical result in the round bracket, and the alphabetical person's significant difference inequality (a=0.05) of same column is identical or do not have the alphabetic flag person's difference is not remarkable.6. immunostimulant is to the influence of porcine blood serum sterilizing function
| Blood sampling time (n=68) | |||||||
| Handle | Before the injection | After the injection | |||||
| 0 day | 3 days | 7 days | 10 days | 14 days | 21 days | 28 days | |
| T | ?0.168±0.044 | ?0.058±0.028 ????(b) | 0.078±0.064 ????(b) | 0.064±0.027 ????(b) | 0.063±0.007 ????(c) | 0.048±0.004 ????(c) | 0.021±0.004 ????(c) |
| B | ?0.168±0.044 | ?0.102±0.025 ????(a) | 0.089±0.019 ????(b) | 0.071±0.038 ????(b) | 0.067±0.020 ????(c) | 0.072±0.024 ????(c) | 0.020±0.018 ????(c) |
| TC | ?0.168±0.044 | ?0.041±0.025 ????(b) | 0.068±0.051 ????(b) | 0.080±0.042 ????(b) | 0.204±0.032 ????(b) | 0.148±0.040 ????(b) | 0.147±0.038 ????(b) |
| BC | ?0.168±0.044 | ?0.092±0.005 ????(a) | 0.157±0.045 ????(a) | 0.186±0.044 ????(a) | 0.353±0.073 ????(a) | 0.300±0.065 ????(a) | 0.278±0.047 ????(a) |
| C | ?0.168±0.044 | ?0.048±0.031 ????(b) | 0.058±0.011 ????(b) | 0.084±0.028 ????(b) | 0.179±0.045 ????(b) | 0.136±0.039 ????(b) | 0.180±0.026 ????(b) |
| N | ?0.168±0.044 | ?0.107±0.090 ????(a) | 0.067±0.049 ????(b) | 0.071±0.004 ????(b) | 0.047±0.020 ????(c) | 0.020±0.022 ????(c) | 0.020±0.015 ????(c) |
The results are shown in Table 7.N group serum is in detection period, and is not obvious to coccus bactericidal actions such as staphylococcus aureus performance, all performance bactericidal actions in various degree after immunity of TC, BC, T, B group.In the time of 3 days, this function performance is the most obvious in immunity, and wherein TC, BC group is extremely remarkable than each matched group difference, but two group differences are not remarkable; TC, BC group and T, B group are significant difference, and the serum sterilizing effect performance of C group is not obvious.Table 7 immunostimulant is to the result that influences of porcine blood serum sterilizing function
Annotate: (1) T: calf thymus element; B: bacillus calmette-guerin vaccine; TC: calf thymus element+swine Fever Vaccine; BC: bacillus calmette-guerin vaccine+swine Fever Vaccine; C: hog cholera immune contrast; N: blank normal healthy controls; NC: normal bacterium number contrast.(2) numeral be x ± SE in the table, and the new multipole difference of The data Deng Kenshi method of inspection (DMRF method) carries out multiple comparisons, and letter is statistical result in the round bracket, and the alphabetical person's significant difference inequality (a=0.05) of same column is identical or do not have the alphabetic flag person's difference is not remarkable.7. the mutual immunoprotection test of rabbit body
| Blood sampling time | |||
| Handle | Before the injection | After the injection | |
| 0 day | 3 days | 7 days | |
| ??T | ????146±14 | ????130±8 ????(b) | ????141±8 ????(a) |
| ??B | ????146±14 | ????123±6 ????(b) | ????132±4 ????(a) |
| ??TC | ????146±14 | ????83±13 ????(c) | ????138±7 ????(a) |
| ??BC | ????146±14 | ????99±13 ????(c) | ????96±21 ????(b) |
| ??C | ????146±14 | ????170±6 ????(a) | ????175±4 ????(a) |
| ??N | ????146±14 | ????149±13 ????(a) | ????169±20 ????(a) |
| ??NC | ????146±14 | ????156±23 ????(a) | ????142±4 ????(a) |
The results are shown in Table 8.The body temperature reaction does not appear in normal healthy controls.Experimental control is behind the hog cholera lapinised virus of 100 ID50 of inoculation; react in the body temperature that 60h occurs up to 41 ℃: do not occur the body temperature reaction except that the BC group has a rabbit; the body temperature reaction of 1 ℃ of normal body temperature all appears exceeding in T, B, each group of C, N, shows that BC group serum antibody may have more protection than all the other each groups.And serum 0.5 of each group of T, B, C, N can't in and the hog cholera lapinised virus of 100 ID50, but have certain difference on the phase in virus lays dormant.The mutual rabbit epidemic disease of table 8 rabbit body protection test result
Annotate: (1) T: calf thymus element; B: bacillus calmette-guerin vaccine; TC: calf thymus element+swine Fever Vaccine; BC: bacillus calmette-guerin vaccine+swine Fever Vaccine; C: hog cholera immune contrast; N: blank normal healthy controls; CC: hog cholera lapinised virus contrast; NN: blank.
| Handle | ????T | ????B | ????TC | ????BC | ????C | ????N | ????CC | ????NN |
| The body temperature reaction | ????+ | ????+ | ????+ | ????+ | ????+ | ????+ | ????+ | ????- |
| ????+ | ????+ | ????+ | ????- | ????+ | ????+ | ????+ | ????- |
(2) the body temperature reaction appears in "+" expression; The body temperature reaction does not appear in "-" expression.
Embodiment 2: bacillus calmette-guerin vaccine is as the immunostimulant of vaccine to chicken coccidiosis.
1, material and method:
Test chicken: the Avian commercial chicken of 1 age in days, under the environment of strict sterilization, raise, feeds utilized for not containing the broiler perfect compound feed of anticoccidial drug.
Test worm strain: tender Eimeria (Eimeria tenella) Wisconsin strain, breed with the preceding healthy chicken that infects through non-ball worm that to test required egg sac number standby.
Immunostimulant: lyophilized bacillus calmette-guerin vaccine, face with before, by every of explanation with 1ml sterilized water for injection dissolving, in the 30min with finishing.
Enzyme labelled antibody: the anti-chicken IgG monoclonal antibody (HRP-McAb) of horseradish peroxidase-labeled: standard operation concentration is 1: 4000.The IgG antibody (HRP-RaCM) of the anti-chicken IgM of the rabbit of horseradish peroxidase-labeled: standard operation concentration is 1: 2000.
Dissolubility spore egg capsule antigen: with reference to the method preparation of (1986) such as Onaga.The E.tenella spore egg capsule of purification is suspended in the phosphate buffer of pH7.6,-20 ℃ with room temperature under multigelation, carry out the Ultrasonic Pulverization cracking under the condition of ice bath, repeatable operation like this, until in test under microscope the time after the complete cracking of egg capsule and zygoblast with the centrifugal 30min of 10000 * g, get-20 ℃ of preservations after the supernatant packing.It is 0.37mg/ml that solubility spore egg capsule antigen records its protein content by ultraviolet spectrophotometry.
Measure lysozyme content in the serum: get serum 10 μ l to be checked, add in the detection hole on the agar plate, put into wet box, hatch 20h for 37 ℃.Measure the diameter of bacteriolyze ring, can obtain lysozyme content in the serum in the substitution standard curve equation.
The mensuration of gram feces egg sac number (OPG): every afternoon, (all around and middle) takes respectively to organize the fresh excreta that chicken was discharged the same day respectively from different positions, and each position is got 3g approximately, adds saturated brine after the weighing, stirs diffusingly, filters; After filtrate shakes up, draw on a small quantity, on McMaster egg count device, count, obtain the OPG value.
Anticoccidial index (ACI) is measured: when 21 ages in days each is organized test chicken by only weighing, eliminate excessive and too small chicken, every group is stayed 10; The worm amount of attacking peroral infection spore egg capsule in accordance with regulations.Infect the back and note observing, cut open inspection after dead chicken is weighed, the record cause of death.Slaughter test chicken after infecting 7.5d,, press Johnson﹠amp by only claiming the end heavy; The method of Reid (1970) is carried out caecum lesion and is kept the score; The night blindness intestinal is respectively organized in collection and content carries out the egg capsule counting.
ACI=(the relative weight gain rate+survival rate)-(pathological changes value+egg capsule is kept the score) 2, EXPERIMENTAL DESIGN: test 1: immunostimulant is established 2 different horizontal dosage respectively to the using dosage, service time and the access times that influence BCG and APS of chicken coccidiosis bar epidemic disease protection: establish 0.1ml/, 0.05ml/ 2 kinds of dosage, APS establishes 1ml/, 0.5ml/ 2 kinds of dosage.Time: divide and immunity while and back 3 days 2 kinds of times of injection of immunity.Number of times: establish injection 1 time and 22 kinds of usages.
The administering mode of BCG and APS is the cervical region subcutaneous injection.The combination of above different usages, every kind of immunostimulant has 8 kinds respectively, need establish 2 * 8 test group; Establish immunity again without reinforcing agent group, not immune worm group, normal healthy controls group, totally 19 groups of attacking.
285 7 age in days Avian chickens are divided into 19 groups at random, 15 every group, use immunostimulant and carry out immunity to coccidiosis by above design.The immune programme for children of coccidiosis (following test together) is: 1000/in 7 age in days per oral inoculation E.tenella spore egg capsules, 14 ages in days inoculate 5000/.250,000 egg capsules during 21 ages in days/only, measure ACI.Test 2: immunostimulant is to the influence of chicken immunity to coccidiosis index
If 6 test group: 1, BCG+ immune group (BI) group; 2, single with BCG immune group (B) group not; 3, APS+ immune group (AI) group; 4, single with APS immune group (A group) not; 5, immunity is without immunostimulant group (I group); 6, normal healthy controls group (C group).The usage of BCG and APS is got the best person of effect in the test 1.
During 7 ages in days, in 302 test chickens, get 20 at random, discard after the blood sampling; All the other are divided into 6 groups at random, 47 every group, use immunostimulant and carry out immunity to coccidiosis by above design.During 21 ages in days with each group test chicken by only weighing, eliminate excessive and too small chicken, every group is stayed 30, is divided into 3 groups (10/group) at random, does not wherein attack worm for 1 group, attacks worm 50,000 and 250,000 egg capsules/only, mensuration ACI respectively for 2 groups in addition.
Remaining blood sampling age in days is: 14,21,28 ages in days, adopt 10 (28 ages in days are institute balance live chickens) at random for every group, and wherein the 14 ages in days chicken of adopting blood also all discards.The index that institute's blood-sample withdrawal is measured has: lymphocyte transformation level, E-rosette forming rate, serum LSZ content, serum IgG and IgM level.In addition, from 7 ages in days, measure the OPG value of each group.Test 3: immunostimulant BCG promotes the discussion of immunity to coccidiosis using method
From APS and BCG, choose immunoenhancement result BCG preferably, study it at lower dosage level (establishing 3 dosage) with the effect when using (use during respectively at 1,4,7 ages in days, in advance 7,3 promptly, 0d) in advance.Have 9 kinds of combinations (9 test group), establish immunity simultaneously without reinforcing agent group, not immune worm group, the normal healthy controls group of attacking.Totally 12 groups.168 1 age in days Avian chickens are divided into 12 groups at random, 14 every group, use immunostimulant and carry out immunity to coccidiosis by above design.During 21 ages in days, measure ACI, attack the worm amount and be 250,000 egg capsules/only.3, result: 3.1 immunostimulants promote the action effect of different immunostimulant effect comparison sheet 1 BCG of the mensuration 3.1.1 of immune protective effect and APS
Annotate: BCG refers to bacillus calmette-guerin vaccine.
| Handle | Dosage (ml/ only) | Inject time (age in days) | Number of times (inferior) | Weightening finish (gram) | The relative weight gain rate (%) | Survival rate (%) | Average lesion score | Egg sac number (* 10 6) | Egg capsule is kept the score | ??ACI |
| ??BCG | ??0.1 | ????7 | ??1 | ??386 | ??124.9 | ??100 | ??1 | ??0.61 | ????1 | ??213.9 |
| ??BCG | ??0.1 | ????7 | ??2 | ??354 | ??114.6 | ??100 | ??0.8 | ??0.075 | ????0 | ??206.6 |
| ??BCG | ??0.1 | ??10 | ??1 | ??360 | ??116.5 | ??100 | ??0.85 | ????0 | ????0 | ??208 |
| ??BCG | ??0.1 | ??10 | ??2 | ??330 | ??106.8 | ??100 | ??1.25 | ????0.45 | ????1 | ?193.3 |
| ??BCG | ??0.05 | ????7 | ??1 | ??333 | ??107.8 | ??100 | ??1.6 | ??0.025 | ????0 | ??191.8 |
| ??BCG | ??0.05 | ????7 | ??2 | ??268 | ??86.7 | ??100 | ??0.75 | ????1.4 | ????5 | ??174.2 |
| ??BCG | ??0.05 | ????10 | ??1 | ??319 | ??103.2 | ??100 | ??0.75 | ????1.35 | ????5 | ?190.7 |
| ??BCG | ??0.05 | ????10 | ??2 | ??304 | ??98.4 | ??100 | ??1.28 | ????0.28 | ????1 | ??184.6 |
| ??APS | ????1 | ????7 | ??1 | ??361 | ??116.8 | ??100 | ??0.55 | ????0.625 | ????1 | ??210.3 |
| ??APS | ????1 | ????7 | ??2 | ??321 | ??103.9 | ??100 | ??0.95 | ????0.125 | ????1 | ??193.4 |
| ??APS | ????1 | ????10 | ??1 | ??325 | ??105.2 | ??100 | ??0.7 | ????0.075 | ????0 | ??198.2 |
| ??APS | ????1 | ????10 | ??2 | ??302 | ??97.7 | ??100 | ??1.25 | ????0.325 | ????1 | ??184.2 |
| ??APS | ????0.5 | ????7 | ??1 | ??271 | ??87.7 | ??100 | ??1.2 | ????0.8 | ????1 | ??174.7 |
| ??APS | ????0.5 | ????7 | ??2 | ??265 | ??85.8 | ??100 | ??1.7 | ????2.025 | ????5 | ??163.8 |
| ?APS | ????0.5 | ????10 | ??1 | ??315 | ??101.9 | ??100 | ??1.5 | ????0.45 | ????1 | ??185.9 |
| ?APS | ????0.5 | ????10 | ??2 | ??299 | ??96.8 | ??100 | ??1.45 | ????0.52 | ????1 | ??181.3 |
| Immunity is without the reinforcing agent group | ??291 | ??94.2 | ??90 | ??2.1 | ????0.78 | ????1 | ??162.2 | |||
| The not immune worm matched group of attacking | ??179 | ??57.9 | ??70 | ??3.55 | ????7 | ????20 | ????72.4 | |||
| The normal healthy controls group | ??309 | ??100 | ??100 | ??0 | ????0 | ????0 | ????200 |
After 17 immune group were attacked worm, every indexs such as the relative weight gain rate, survival rate, pathological changes value and egg capsule value all obviously were better than the not immune worm group of attacking, and illustrate that the immune programme for children that is adopted is successfully effective.Immunity is 162.2 without the ACI of reinforcing agent group, uses BCG can improve the anticoccidial index to some extent.The relative weight gain rate of 8 groups of injection of BCG is 86.7% except that one group, and all the other each groups are higher than immunity without 94.2% of reinforcing agent group between 98.4～124.9%; Pathological changes is light (0.75～1.28); ACI is 174.2～213.9, apparently higher than immunity without the reinforcing agent group.The action effect of APS is then poor slightly, but most group is better than immunity without the reinforcing agent group; The relative weight gain is 85.8-116.8%, and lesion score is 0.55-1.7, and ACI163.8-210.33.1.2 attacks the immune protective effect of immunostimulant under the worm intensity in difference
The results are shown in Table 2, when attacking worm 50,000 spore egg capsules, the relative weight gain rate of BI group reaches 1 14.5%; ACI is 210.5; be higher than all that (the relative weight gain rate 106.0% ACI=201.5), shows that chicken has obtained to protect fully to the attack of 50,000 spore egg capsules without the I of immunostimulant group.250,000 spore egg capsules are attacked, and the relative weight gain rate of BI group and two indexs of lesion score also are better than the I group without immunostimulant, and ACI reaches 203.0, is higher than I group (ACI=177.7), and the BCG action effect is quite obvious.
The single infection that then can not resist 50,000 or 250,000 spore egg capsules with the not immune B group of immunostimulant, weightening finish is obstructed, and pathological changes is serious; ACI is 137.6 when 50,000 egg capsules are attacked worm, and it is then lower to attack worm 250,000 egg capsules. only 114.0, and invalid substantially.Table 2 difference is attacked the ACI measurement result of worm amount
| Group number | Attack worm amount (* 10 4) | Weightening finish (gram) | The relative weight gain rate (%) | Survival rate (%) | Average lesion score | Egg sac number (* 10 6) | Egg capsule is kept the score | ??ACI |
| ??C-1 | ????0 | ??385 | ??100 | ??100 | ????0 | ????0 | ????0 | ??200 |
| ??C-2 | ????5 | ??308 | ??80 | ??100 | ??2.95 | ??10.1 | ??30 | ??120.5 |
| ??C-3 | ????25 | ??228 | ??59.2 | ???80 | ??3.75 | ??9.5 | ??20 | ??81.7 |
| ??I-1 | ????0 | ??404 | ??104.9 | ??100 | ||||
| ??I-2 | ????5 | ??408 | ??106.0 | ??100 | ??0.35 | ??0.2 | ????1 | ??201.5 |
| ??I-3 | ????25 | ??376 | ???97.7 | ??100 | ???1.9 | ???0.7 | ????1 | ??177.7 |
| ??BI-1 | ????0 | ??451 | ??117.1 | ??100 | ||||
| ?AI-1 | ????0 | ??471 | ??122.3 | ??100 | ||||
| ?AI-2 | ????5 | ??459 | ??119.2 | ??100 | ??0.25 | ??0.15 | ????1 | ??215.7 |
| ??AI-3 | ????25 | ??438 | ??113.8 | ??100 | ????1 | ??0.93 | ????1 | ??202.8 |
| ??A-1 | ????0 | ??422 | ??109.6 | ??100 | ||||
| ??A-2 | ????5 | ??361 | ??93.8 | ??100 | ??2.6 | ??11.4 | ????40 | ??127.8 |
| ??A-3 | ????25 | ??250 | ??64.9 | ???80 | ??3.25 | ???8.7 | ????20 | ??92.4 |
| ??BI-2 | ????5 | ??441 | ??114.5 | ??100 | ??0.3 | ??0.25 | ????1 | ??210.5 |
| ??BI-3 | ????25 | ??422 | ??109.6 | ??100 | ??0.56 | ???0.5 | ????1 | ??203.0 |
| ??B-1 | ????0 | ??418 | ??108.6 | ??100 | ||||
| ??B-2 | ????5 | ??397 | ??103.1 | ??100 | ??2.55 | ??11.6 | ??40 | ??137.6 |
| ??B-3 | ????25 | ??260 | ??67.5 | ???90 | ??3.35 | ???4.2 | ??10 | ??114 |
Annotate: the numeral behind the letter is a grouping serial number.C: normal healthy controls; I: immunity is without reinforcing agent; AI: astragalus polysaccharides+immune A: single astragalus polysaccharides of using, astragalus polysaccharides 0.5ml/, BI: bacillus calmette-guerin vaccine+immunity; B: single not immune with bacillus calmette-guerin vaccine.Injection is 1 time during 7 ages in days.The bacillus calmette-guerin vaccine usage is 0.05ml/, and injection is 1 time during 7 ages in days.3.1.3 BCG promotes the selection of immune optimum condition
The results are shown in Table 3, use 9 test group of BCG all to have certain effect on raising the relative weight gain rate, reduction lesion score, ACI is between 175.3-202.6; Wherein best in 1 age in days injector effect with 0.025ml/ dosage only, the relative weight gain rate reaches 116.1%, lesion score only 0.85 minute, and ACI is 202.6; And attack worm group ACI only 165.8 without the immunity of BCG.Adopt L9 (3
4) orthogonal table carries out statistical analysis and significance test to data, the result is that the size of dosage is the main factor that influences the ACI height, secondly is the injection age in days.0.025ml/ of use or 0.05ml/ dose effect only are all relatively good; Immunity was used the effect of BCG in preceding 7 days and is shifted to an earlier date 3 days or use no significant difference simultaneously.The effect of the different usages of table 3 BCG relatively
Annotate: BCG refers to bacillus calmette-guerin vaccine 3.2 cellular immunization index determining 3.2.1 lymphocyte transformation levels
| Handle | Dosage (ml/ only) | Inject time (age in days) | Weightening finish (gram) | The relative weight gain rate (%) | Survival rate (%) | Average lesion score | Egg sac number (* 10 6) | Egg capsule is kept the score | ??ACI |
| ?BCG | ?0.05 | ?1 | ?444 | ?109.9 | ?100 | ?0.95 | ?0.7 | ?1 | ?199.4 |
| ?BCG | ?0.05 | ?4 | ?436 | ?107.9 | ?100 | ?1.35 | ?1.45 | ?5 | ?189.4 |
| ?BCG | ?0.05 | ?7 | ?435 | ?107.7 | ?100 | ?1.2 | ?0.9 | ?1 | ?194.7 |
| ?BCG | ?0.025 | ?1 | ?469 | ?116.1 | ?100 | ?0.85 | ?1.7 | ?5 | ?202.6 |
| ?BCG | ?0.025 | ?4 | ?433 | ?107.2 | ?100 | ?1.35 | ?0.8 | ?1 | ?192.7 |
| ?BCG | ?0.025 | ?7 | ?409 | ?101.2 | ?100 | ?1 | ?0.75 | ?1 | ?190.2 |
| ?BCG | ?0.0125 | ?1 | ?408 | ?101.0 | ?100 | ?1.55 | ?1.5 | ?5 | ?180.6 |
| ?BCG | ?0.0125 | ?4 | ?377 | ?93.3 | ?100 | ?1.3 | ?1.8 | ?5 | ?175.3 |
| ?BCG | ?0.0125 | ?7 | ?397 | ?98.3 | ?100 | ?1.75 | ?1.15 | ?5 | ?175.8 |
| Immunity is without reinforcing agent | ?367 | ?90.8 | ?100 | ?2.5 | ?1.0 | ?1 | ?165.8 | ||
| Not immunely attack worm contrast | ?204 | ?50.5 | ?80 | ?3.75 | ?6.375 | ?20 | ?73 | ||
| Normal healthy controls | ?404 | ?100 | ?100 | ?0 | ?0 | ?0 | ?200 |
The results are shown in Table 4, the lymphocyte transformation ability was very low when C organized 7 ages in days, along with the increase meeting of age in days raises gradually.1 group similar to the C group, and difference is not remarkable.That uses APS and BCG respectively organizes the OD value all than 1 group and C class value height, significant difference.After attacking worm, it is all less that each organizes the variation of OD value.Table 4 lymphocyte transformation test result (OD
570)
Illustrate: (1) C: normal healthy controls; I: immunity is without reinforcing agent; AI: astragalus polysaccharides+immune A: single astragalus polysaccharides, astragalus polysaccharides usage used is 0.5ml/, BI: bacillus calmette-guerin vaccine+immunity; B: single not immune with bacillus calmette-guerin vaccine.Injection is 1 time during 7 ages in days.The bacillus calmette-guerin vaccine usage is 0.05ml/, and injection is 1 time during 7 ages in days.
| Group | 7 ages in days | 14 ages in days | 21 ages in days | 28 ages in days | ||
| Do not attack worm | Attack worm 50,000 | Attack worm 250,000 | ||||
| The C group | 0.122±0.009 | ??0.150±0.015 ????(c) | ??0.208±0.022 ????(c) | 0.232±0.018 ????(c) | 0.231±0.016 ????(c) | 0.210±0.029 ????(c) |
| The I group | 0.122±0.009 | ??0.149±0.017 ????(c) | ??0.238±0.018 ????(c) | 0.214±0.014 ????(c) | 0.251±0.022 ????(c) | 0.237±0.029 ????(c) |
| The BI group | 0.122±0.009 | ??0.302±0.026 ????(a) | ??0.834±0.048 ????(a) | 0.779±0.048 ????(a) | 0.818±0.054 ????(a) | 0.861±0.076 ????(a) |
| The B group | 0.122±0.009 | ??0.265±0.016 ????(ab) | ??0.721±0.045 ????(b) | 0.689±0.044 ????(a) | 0.703±0.047 ????(ab) | 0.725±0.066 ????(ab) |
| The AI group | 0.122±0.009 | ??0.271±0.016 ????(ab) | ??0.774±0.038 ????(ab) | 0.744±0.045 ????(a) | 0.789±0.065 ????(a) | 0.794±0.060 ????(a) |
| The A group | 0.122±0.009 | ??0.242±0.014 ????(b) | ??0.666±0.035 ????(b) | 0.561±0.039 ????(b) | ??0.602±0.041 ????(b) | 0.587±0.056 ????(b) |
(2) numeral be X ± SE in the table, and multiple comparisons adopts the newly heavy range test method (DMRT method) of Deng Kenshi, and letter is statistical result in the round bracket, and the alphabetical person's significant difference inequality (a=0.05) of same column is identical or do not have the alphabetic flag person's difference is not remarkable.3.2.2 E-rosette forming rate
The results are shown in Table 5,14,21 ages in days, the E-rosette forming rate of BI group all is significantly higher than other each group, significant difference; The E-rosette forming rate of B group, A group and I group is also higher, organizes significant difference with C during 21 ages in days.28 ages in days, the E-rosette forming rate of not attacking each group of worm all has in various degree decline, but still is height with BI group, AI group; After attacking worm, the E-rosette forming rate of non-immune B group, A group and C group raises, and the BI of immunity group, AI group and I group, except that I organizes attacking 250,000 egg capsules of a worm/back E-rosette forming rate rising, all the other each groups still descend, but decline scope is less than attacking worm group, and still keeps higher level after attacking 250,000 egg capsules of worm/only.Table 5 E-rosette result (%)
Illustrate: (1) C: normal healthy controls; I: immunity is without reinforcing agent; AI: astragalus polysaccharides+immune A: single astragalus polysaccharides, astragalus polysaccharides usage used is 0.5ml/, BI: bacillus calmette-guerin vaccine+immunity; B: single not immune with bacillus calmette-guerin vaccine.Injection is 1 time during 7 ages in days.The bacillus calmette-guerin vaccine usage is 0.05ml/, and injection is 1 time during 7 ages in days.
| Group | 7 ages in days | 14 ages in days | 21 ages in days | 28 ages in days | ||
| Do not attack worm | Attack worm 50,000 | Attack worm 250,000 | ||||
| The C group | 11.56±0.64 | ??15.5±0.97 ????(c) | ??19.46±1.07 ????(d) | ??20.7±1.25 ????(c) | ??25.74±1.22 ????(c) | ??23.56±1.74 ????(c) |
| The I group | 11.56±0.64 | ??18.11±0.84 ????(bc) | ??24.38±1.38 ????(c) | ??21.67±1.41 ????(bc) | ??23.56±0.96 ????(bc) | ??28.13±1.08 ????(ab) |
| The BI group | 11.56±0.64 | ??24.18±1.09 ????(a) | ??35.04±1.18 ????(a) | ??27.18±1.19 ????(a) | ??26.46±0.99 ????(abc) | ??31.30±1.18 ????(a) |
| The B group | 11.56±0.64 | ??18.64±1.34 ????(bc) | ??28.62±1.29 ????(b) | ??23.76±1.22 ????(abc) | ??30.01±0.91 ????(a) | ??26.83±1.46 ????(bc) |
| The AI group | 11.56±0.64 | ??24.67±1.34 ????(a) | ??32.87±1.18 ????(a) | ??25.79±1.19 ????(a) | ??28.00±1.28 ????(ab) | ??30.18±1.27 ????(ab) |
| The A group | 11.56±0.64 | ??20.47±1.59 ????(b) | ??27.07±1.31 ????(bc) | ??24.91±1.15 ????(ab) | ??29.56±2.12 ????(ab) | ??27.50±1.62 ????(abc) |
(2) numeral be X ± SE in the table, and multiple comparisons adopts the newly heavy range test method (DMRT method) of Deng Kenshi, and letter is statistical result in the round bracket, and the alphabetical person's significant difference inequality (a=0.05) of same column is identical or do not have the alphabetic flag person's difference is not remarkable.3.2.3 serum LSZ assay
The results are shown in Table 6,7～28 ages in days and respectively organize all constantly increases of LSZ content of test chicken, BCG can increase serum LSZ content.14 ages in days, the LSZ content of A group reaches 12.5 μ g/m1; 21 ages in days, the BI group is 18.35 μ g/ml, the AI group is 17.8 μ g/ml, all is significantly higher than C group and I group.Table 6 serum LSZ assay result (μ g/ml)
Illustrate: (1) C: normal healthy controls; I: immunity is without reinforcing agent; AI: astragalus polysaccharides+immune A: single astragalus polysaccharides, BI used: bacillus calmette-guerin vaccine+immunity; B: single not immune with bacillus calmette-guerin vaccine.The astragalus polysaccharides usage is 0.5ml/, and injection is 1 time during 7 ages in days.The bacillus calmette-guerin vaccine usage is 0.05m1/, and injection is 1 time during 7 ages in days.
| Group | 7 ages in days | 14 ages in days | 21 ages in days | 28 ages in days | ||
| Do not attack worm | Attack worm 50,000 | Attack worm 250,000 | ||||
| The C group | 9.12±0.39 | ??9.49±0.38 ????(b) | ??11.11±1.16 ????(b) | ??13.88±0.76 | ??13.87±1.27 | ??12.28±1.53 ????(c) |
| The I group | 9.12±0.39 | ??9.35±0.41 ????(b) | ??11.95±2.21 ????(b) | ??12.46±1.16 | ??13.31±0.80 | ??12.64±1.54 ????(bc) |
| The BI group | 9.12±0.39 | ??10.19±0.90 ????(b) | ??18.35±1.17 ????(a) | ??14.83?±0.96 | ??15.33±1.20 | ??17.84±0.88 ????(a) |
| The B group | 9.12±0.39 | ??9.73±0.57 ????(b) | ??15.56±2.27 ????(ab) | ??15.26±1.22 | ??15.73±1.21 | ??16.16±1.05 ????(ab) |
| The AI group | 9.12±0.39 | ??10.79±0.40 ????(ab) | ??17.8±1.75 ????(a) | ??16.27±1.51 | ??17.29±1.18 | ??15.75±0.91 ????(abc) |
| The A group | 9.12±0.39 | ??9.73±0.57 ????(a) | ??9.73±0.57 ????(ab) | ??9.73±0.57 | ??9.73±0.57 | ??9.73?±0.57 ????(abc) |
(2) numeral be X ± SE in the table, and multiple comparisons adopts the newly heavy range test method (DMRT method) of Deng Kenshi, and letter is statistical result in the round bracket, and the alphabetical person's significant difference inequality (a=0.05) of same column is identical or do not have the alphabetic flag person's difference is not remarkable.3.3 humoral immunization index: the dynamic change of serum IgG, IgM assay 3.3.1 serum IgG
The results are shown in Table 7, entire test, the OD value of AB, C group is lower than the positive reaction starting point.Al, BI, I organize when 21 ages in days, i.e. immunity is 2 weeks afterwards, can detect the antibody response of higher level in the serum, and wherein the OD value of BI group is higher than Al, I group, significant difference.28 ages in days, OD value still continue to raise, and can excite higher levels of antibody to produce after attacking worm.ELISA measurement result (the OD of IgG level in table 7 serum
490)
Illustrate: (1) C: normal healthy controls; I: immunity is without reinforcing agent; AI: astragalus polysaccharides+immune A: single astragalus polysaccharides, BI used: bacillus calmette-guerin vaccine+immunity; B: single not immune with bacillus calmette-guerin vaccine.The astragalus polysaccharides usage is 0.5ml/, and injection is 1 time during 7 ages in days.The bacillus calmette-guerin vaccine usage is 0.05ml/, and injection is 1 time during 7 ages in days.
| Group | 7 ages in days | 14 ages in days | 21 ages in days | 28 ages in days | ||
| Do not attack worm | Attack worm 50,000 | Attack worm 250,000 | ||||
| The C group | 0.26±0.013 | ??0.221±0.009 | ?0.207±0.012 ????(c) | ??0.238±0.013 ????(c) | ????0.310±0.016 ????(c) | ????0.283±0.017 ????(c) |
| The I group | 0.26±0.013 | ?0.293±0.016 | ?0.601±0.038 ????(b) | ??0.733±0.049 ????(b) | ????0.862±0.047 ????(b) | ????0.897±0.055 ????(b) |
| The BI group | 0.26±0.013 | ??0.274±0.020 | ?0.812±0.061 ????(a) | ??0.942±0.055 ????(a) | ????1.002±0.053 ????(a) | ????1.037±0.064 ????(a) |
| The B group | 0.26±0.013 | ??0.245±0.011 | ??0.253±0.014 ????(c) | ??0.218±0.011 ????(c) | ????0.264±0.022 ????(c) | ????0.301±0.019 ????(c) |
| The AI group | 0.26±0.013 | ??0.246±0.017 | ??0.66±0.041 ????(b) | ??0.709±0.039 ????(b) | ????0.812±0.048 ????(b) | ???0.927±0.045 ????(ab) |
| The A group | 0.26±0.013 | ??0.198±0.012 | ??0.235±0.011 ????(c) | ??0.224±0.014 ????(c) | ????0.286±0.012 ????(c) | ????0.276±0.023 ????(c) |
(2) numeral be X ± SE in the table, and multiple comparisons adopts the newly heavy range test method (DMRT method) of Deng Kenshi, and letter is statistical result in the round bracket, and the alphabetical person's significant difference inequality (a=0.05) of same column is identical or do not have the alphabetic flag person's difference is not remarkable.3.3.2 the dynamic change of serum IgM
The serum OD value that the results are shown in Table 8,7～21 age in days A, B, C group all is lower than the positive reaction starting point, but attacks the IgM that can measure higher level behind the worm when 28 ages in days.Antibody horizontal difference between three groups is not remarkable.
3 immune group (Al, BI, I group) when 14 ages in days, i.e. in 1 week after the immunity, can detect a certain amount of IgM antibody in the serum, rise to the peak during 21 ages in days, and 28 ages in days are reduced to lower level.Though attack the downward trend that worm fails to change IgM, IgM can keep higher level.Duration of test, the OD value of BI group are higher than I group and AI group, but difference is not remarkable.ELISA measurement result (the OD of table 8 serum IgM level
490)
Illustrate: (1) C: normal healthy controls; I: immunity is without reinforcing agent; Al: astragalus polysaccharides+immune A: single astragalus polysaccharides, BI used: bacillus calmette-guerin vaccine+immunity; B: single not immune with bacillus calmette-guerin vaccine.The astragalus polysaccharides usage is 0.5ml/, and injection is 1 time during 7 ages in days.The bacillus calmette-guerin vaccine usage is 0.05ml/, and injection is 1 time during 7 ages in days.
| Group | 7 ages in days | 14 ages in days | 21 ages in days | 28 ages in days | ||
| Do not attack worm | Attack worm 50,000 | Attack worm 250,000 | ||||
| The C group | 0.298±0.012 | ??0.295±0.060 ????(c) | ??0.285±0.016 ????(b) | 0.255±0.009 ????(c) | 0.611±0.033 | ?0.654±0.047 |
| The I group | 0.298±0.012 | ??0.576±0.028 ????(b) | ??0.804±0.047 ????(a) | 0.554±0.040 ????(ab) | 0.592±0.037 | ?0.704±0.060 |
| The BI group | 0.298±0.012 | ??0.691±0.048 ????(b) | ??0.853?±0.044 ????(a) | 0.604±0.052 ????(a) | 0.664±0.061 | ?0.739±0.050 |
| The B group | 0.298±0.012 | ??0.259±0.015 ????(c) | ??0.269±0.016 ????(b) | 0.265±0.011 ????(c) | 0.544±0.023 | ?0.607±0.045 |
| The AI group | 0.298±0.012 | ??0.624±0.031 ????(ab) | ??0.765±0.047 ????(a) | 0.482±0.029 ????(b) | 0.623±0.039 | ?0.669±0.068 |
| The A group | 0.298±0.012 | ??0.269±0.014 ????(c) | ??0.302±0.010 ????(b) | 0.279±0.011 ????(c) | 0.567±0.044 | ?0.666±0.064 |
(2) numeral be X ± SE in the table, and multiple comparisons adopts the newly heavy range test method (DMRT method) of Deng Kenshi, and letter is statistical result in the round bracket, and the alphabetical person's significant difference inequality (a=0.05) of same column is identical or do not have the alphabetic flag person's difference is not remarkable.
Claims (5)
1, a kind of vaccine immunity of animals is played the bacillus calmette-guerin vaccine injecting method of potentiation, it is characterized in that: bacillus calmette-guerin vaccine is injected on one's body the animal for the treatment of immunity inoculation before vaccine injection or with vaccine simultaneously.
2, method according to claim 1 is characterized in that: the animal for the treatment of immunity inoculation is the young pig in 20～40 day age, and the bacillus calmette-guerin vaccine injection volume is 0.1～0.2 a milligram/head.
3, method according to claim 1 is characterized in that: the animal for the treatment of immunity inoculation is the young chicken in 7～14 day age, and the bacillus calmette-guerin vaccine injection volume is 0.025～0.05 milligram/.
4, method according to claim 2 is characterized in that: vaccine is a swine Fever Vaccine, and the bacillus calmette-guerin vaccine injection is carried out simultaneously with the swine Fever Vaccine inoculation.
5, method according to claim 3 is characterized in that: vaccine is the chicken coccidiosis vaccine, and the bacillus calmette-guerin vaccine injection is carried out simultaneously with the chicken coccidiosis vaccine injection or carried out in 7 days in advance.
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