| Canine distemper, canine parvovirus disease and infectious canine hepatitis are three kinds of important infectious diseases harming dogs seriously. Because they are viral infectious diseases, the vaccination is the most effective measure to control their occurrence. It is very important to enact scientific and rational immunization programme. Some factors directly affect the effect of prevention and control, such as the level of basic antibody level, the first-vaccination age of young dog, vaccine and dose. Xigang canine farm is one of the socialization canine-feeding farms which belongs to Nanjing Institute of Police dog of the Ministry of Public Security. At present, the canine farm carries out the immunization programme from "the working regulation of disease prevention for police dog". Whether these procedures are reasonable or not? A preliminary investigation was carried out. In this study the determination included the changes of serum canine distemper virus (CDV) antibody, canine parvovirus (CPV) antibody, canine adenovirus (CAV) antibody titers and peripheral white blood cell in adult dogs after routine immunization, the effect of the first-vaccination ages, dose and vaccine from different source on three antibody in puppies, and the effect of Chinese herbal medicinal adjuvant on three antibody titers and serum contents of IFN-γ, IL-2, IL-4and IL-10in puppies. The purpose of this study is to select the optimal first-vaccination age, dose, vaccine and adjuvant, and provide the theoretical evidence for establishing scientific and reasonable immune procedure and improving the preventive effect of three kinds of infectious diseases. The details are divided into six parts as follows:Experiment1. The changes of three antibody titers and white blood cell in adult dogs after routine immunization In order to validate the effectiveness of routine vaccination in adult dogs and the effect of basic antibody levels, sex and age on humoral immunity. Five young female dogs (Ⅰ), four older male dogs (Ⅱ) and four young male dogs (III) were selected randomly. All dogs in three groups were injected with Pfizer quadruple vaccine,1head-dose per dog respectively. Before vaccination (Do) and on day15(D15), months1(M1),3(M3),6(M6) and9(M9) after vaccination, blood was sampled for determination the changes of CDV antibody, CPV antibody, CAV antibody titers, white blood cell count and classification. The results showed that the CDV antibody, there were no significant differences among all group at all time points, and presented high-low-high tendency in whole year. The CPV antibody, in three groups displayed high-low-high-low tendency in whole year, and in older male dogs were significantly lower than those in other two groups at four time points. The CAV antibody, also presented high-low-high-low tendency in whole year, and in young male dogs on M6was significantly lower than those in other two groups. The three kinds of antibody titers in young female dogs were at higher level. The total white blood cells in young female dogs presented a larger rise on days15after vaccination and was significantly higher than those in other two groups. The leukocyte differential counts had no significant difference among three groups at all time points. These results indicated that it was feasible and effective for adult dogs to carry out one shot immunity per year according to "the working regulation of disease prevention for police dog". In the older dogs had the complete immune memory response. The antibody level in young female dogs was higher,therefore, the level of maternal antibody should be considered in immune programme in newly born puppies.Experiment2. Effect of first-vaccination age on serum antibody titers of puppy In order to verify whether45-day-old is the most suitable time of first vaccination of puppy or not, the35-day-old (Ⅰ),40-day-old (Ⅱ),50-day-old (Ⅲ) and45-day-old (control, C) dogs,4dogs in each group, were randomly selected and vaccinated with canine distemper-canine parvovirus bivalent vaccine. The re-vaccinations were performed on14and28days after the first vaccination with Intervet quadruple vaccine and Pfizer quadruple vaccine respectively. Before vaccination (Do) and on days7(D7),14(D]4),21(D2)),28(D28),35(D35) and42(D42) after the first vaccination, the blood was sampled for determinating the changes of CDV antibody, CPV antibody and CAV antibody titers. The results showed that CDV antibody, the time points with big extent rise were gradually advanced along with the increasing of age, there was an inverse ratio between the maternal antibody level and the rise extent of antibody on puppy after vaccination, and the first vaccination at50-day-old was best, at45-day-old, better. For CPV antibody, first vaccination at45-day-old was best, at40-day-old, secondary. For CAV antibody, first vaccination at35-day-old was best, at45-day-old, next. In general consider45-day-old was reasonable first-vaccination age.Experiment3. Effect of first-vaccination dose on serum antibody titers of puppy In order to select out the optimal first vaccination dose, sixteen45-day-old puppies were randomly assigned into four groups,4puppies each group and vaccinated with Intervet bivalent vaccine respectively at0.5head-dose (Ⅰ),1.5head-dose (Ⅱ),2.0head-dose (Ⅲ) and1.0head-dose (C), repeated vaccination on days14and28after the first vaccination respectively with Intervet quadruple vaccine and Pfizer quadruple vaccine. Before vaccination (Do) and on days7(D7),14(D14),21(D21),28(D28),35(D35) and42(D42) after the first vaccination, the blood was sampled for determinating the change of serum CDV antibody, CPV antibody and CAV antibody. The results showed that CDV antibody, in0.5head-dose group began to increase on D7and always kept at a high level at the next5time points; the peak value came out earlier than those in other3groups and was higher than those in other3groups reaching81.41%. The CPV antibody, in0.5head-dose group started to increase on D14, which was earlier2weeks than1.5head-dose group, subsequently sustained at high level and the peak value was earlier one week than in1.5head-dose group. The CAV antibody, in0.5head-dose group on D35elevated significantly and reached the peak value with a rise extent of33.86%being close to those in2.0head-dose and1.5head-dose groups, and o D42sustain higher level. These results suggested that0.5head-dose was the the optimal first-vaccination dose.Experiment4. Effect of different sources vaccine at first vaccination on serum antibody titer of puppy In order to select out the optimal vaccine for the first vaccination, sixteen45-day-old puppies were randomly assigned into four groups,4puppies each group, vaccinated respectively with Five-star quintavalent vaccine (Ⅰ), Intervet quadruple vaccine (Ⅱ), Pfizer quadruple vaccine (Ⅲ) and Intervet bivalent vaccine (C), and repeated vaccination on days14and28after the first vaccination respectively with Intervet quadruple vaccine and Pfizer quadruple vaccine. Before vaccination (Do) and on days7(D7),14(D14),21(D21),28(D28),35(D35) and42(D42) after the first vaccination, the blood was sampled for determinating the change of serum CDV antibody, CPV antibody and CAV antibody. The results showed that the CDV antibody, the rise extent of peak value in II group was hight arriving77.85%. The next was C group. CPV antibody, the peak value in C group was highest with a rise extent of56.23%and stable ascension, and still sustained at higher level on D42. The next was Ⅱ group. CAV antibody, the peak value and its rise extent in Ⅲ group were highest (48.67%), the second was Ⅱ group. In general comparison, Intervet quadruple vaccine was better in first vaccination.Experiment5. Effect of Chinese herbal medicinal adjuvant on serum antibody titer of vaccinated puppy In order to study the immunoenhancement of Chinese herbal medicinal adjuvant, twenty-four45-old-day puppies were randomly assigned into6groups (4puppies a group), prescription Ⅰ low dose (Ⅰ L) and high dose (Ⅰ H)group, prescriptions Ⅱ low dose (ⅡL) and high dose (ⅡH) group, levamisole group (adjuvant control, AC) and non-adjuvant control (NA) group. All puppies were vaccinated with Intervet bivalent vaccine, and repeated vaccination on days14and28after the first vaccination respectively with Intervet quadruple vaccine and Pfizer quadruple vaccine. At the same time of every vaccination, puppies in the four Chinese herbal medicine groups were intramuscularly injected1mL of corresponding drug in other side of neck, in AC group, given Ergamisole2.5mg/kg per os, and in NA group not given any drug. Before vaccination (Do) and on days7(D7),14(D14),21(D21),28(D28),35(D35) and42(D42) after first vaccination, blood were sampled from forelimb vein for determination of CDV antibody, CPV antibody and CAV antibody. The results showed that for CDV antibody, the effect of ⅠH was best, which risen sharply from D7, upto D42kept at high level, and the amplification of peak value was82.68%and much higher than the rest groups, the following was ⅡH group. For CPV antibody, the effect of ⅠL was best, the following was ⅠL. This two groups reached peak value respectively on D14and D28, with a amplifications of36.84%and keeping at higher level during D14to D42. For CAV antibody, the effect of ⅠH was best, on D28presentd a larger rise extent and on D42reached peak value with a highest amplification (41.40%), the following was ⅡL group. These results revealed that the Chinese herbal medicinal adjuvant possessed obvious immunoenhancement, and Prescription I expressed a better effect.Experiment6. Effect of Chinese herbal medicinal adjuvant on content of cytokine in vaccinated puppy In order to study the immunoenhancement mechanism of Chinese herbal medicinal adjuvant, Twenty-four45-day-old puppies were selected and treated similar to test5. Before vaccination (Do) and on days 14(D14),21(D21),28(D28),35(D35) and42(D42) after vaccination, blood were sampled from forelimb vein for determination of the change of serum IL-2, IFN-γ, IL-4and IL-10content. The results showed that IL-2content, in Ⅰ H, ⅠL group on D14and four Chinese herbal medicine groups on D28and D42were significantly higher than those in NA group (P<0.05), in ⅡH group on D28was the highest, the next was ⅠL group and they were280.29%and194.03%of those on Do respectively. IFN-y content, in ⅠL group on D14, four Chinese herbl medicine groups on D28,ⅠL, ⅠH and ⅡL group on D42were significantly higher than those in NA group (P<0.05), in ⅡH group on D28was highest, the following was ⅡL group on D42the two groups made up176.30%and136.35%of those on Do, respectively. IL-4content, in ⅠH group on D42was highest, significantly higher than that in NA group (P<0.05) and it made up114.55%of that on Do, the following was ⅡL group on D14made up97.5%of that on on D0.IL-10content, in the four Chinese herbal medicine groups on D14, ⅠH and ⅡH on D28and D42were significantly higher than those in NA group (P<0.05). All these results revealed that Chinese herbal medicinal adjuvant could significantly promote the secretion of cytokine, and both of two prescriptions possessed better effect.Based to above research, the immune programme for three infectious diseases of young dog in our farm was decided:the first vaccination at45-days-old with0.5head-dose of Intervet quadruple vaccine, the second with Intervet quadruple vaccine and third with Pfizer quadruple vaccine after2weeks interval. At the same times each vaccination, high dose of Chinese herbal medicinal adjuvant I was injected. By more than two years’application, the satisfactory effect was obtained for three infectious diseases in our farm. |
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