Studies On The Preparation And Clinical Application Of Long-Acting Diluent For Swine Semen

The preservation of boar semen at room temperature is an important part in artificial insemination, which is important to reduce production cost and to increase economic efficiency. However, the effective time of semen stored at room temperature is only 2 3d normally in our country, and most of the pig farms use the self-assigned diluent, which has a short preserved time. Currently, domestic used boar semen diluent stored at room temperature are mostly purchased from abroad where the prices of imported goods are more expensive, which increasing the cost of breeding, so the development of semen diluent that can keep a long time and preserve in good effect,and the cost of which is significantly lower than the current commercialization has important economic benefits in terms of the artificial insemination technique. The boar semen dilution formulation preserved at room-temperature and the preservation techniques are investigated in this thesis. Starting with the ingredients in the formula, the experiment compared the effects of several additives on the semen storage. By using the orthogonal experimental,the basic formula was obtained and its stability was investigated,and the quality standards was developed, and the comparison is couducted in clinical reproduction test with the similar products in market..The six factors and four-level orthogonal experimental were used for selecting the formula in the study, in which the ratio of semen and diluent was set at 1:2, and the proportion of each components was determined though the determination of semen vitality; setting up different semen dilution ratio after the formula being selected, and determining the semen vitality at 0h,24h,48h,72h,96h,120h respectively to determine the optimum dilution ratio; The stability of semen diluent was examined under the conditions of high, medium and low temperature (4℃,40℃,60℃) and humidity (60%, 45%,28%) by using of accelerated stability test; Adding the semen diluent with different anti-microbial drugs (penicillin+streptomycin, enrofloxacin, Colistin Sulfate, Enrofloxacin+amphotericin B), and setting the ratio of the semen and semen diluent at 1:1,1:2,1:4,1:8, respectively, and examining the preservation vitality to select the appropriate antibiotics under the conditions of 17℃at 0h,24h,48h,72h,96h,120h; Under the same conditions, comparing the preserved effect on semen vitality and preserving time with the imported BTS, ZorPVa semen diluent; establishing an compared trials with the foreign similar products to determine the pregnancy rate (%) and the live number of the semen diluent of artificial insemination. The ratio of glucose, sodium citrate, sodium bicarbonate, EDTA disodium, potassium chloride in semen diluent formulations is selected by orthogonal experiments, which is 0.8:0.13:0.027:0.027:0.016; By the dilution Preservation experiments with different dilution ratio, it is found that the semen has a stronger motility with the ratio for semen and diluent ranged from 1:1 to 2 at he same time. The stability of diluent is studied by setting the high, medium and low temperature and humidity respectively, the results showed that the semen diluent is suitable for being preserved at lower temperature and in the shade, preferably at 4℃in the refrigerator and the humidity at 28% or less; With different antibiotics add experiments,it is obtained that the diluent has higher potency and longer maintenance of semen motility by adding Enrofloxacin+amphotericin B. The effective energy is above 0.7 when the semen diluent is preserved more than 5 days, the preservation effect is better than the BTs and equal to the ZorPVa. The clinical reproduction trials were conducted by taking imported ZorPVa semen powders as references, the results show that the test products and the reference substances were in the same pregnancy rate with 93.3%, the difference was not statistically significant (P> 0.05); the live number of the test group is 9.64 and the reference group of which is 9.71, the difference was not statistically significant (P> 0.05).The semen diluent which can be saved more than 5 days and the effective energy above 0.7 were developed in this thesis, and the research target was achieved. In the next step, it is needed to establish a pilot test base jointed with the bushiness, produce with small-batch, promote and apply in the clinical practice, and conduct a continuous sum-up and improvement.