| Colibacillosis is a kind of frequent, acute, intestinal infectious diseases with high mortality, which was mainly caused by Enterotosigenci Escherichia coli(ETEC) and have a great harm on the growth and development of newborn piglets. With the development of pig breeding industry, the morbidity and mortality of the disease has been increasing. As pig-breeding industry growing, its morbidity rate and mortality rate has been influnced. Through continual work, Chinese Herbal Medicine microorganism fermentation has become a new way to prevent and control the disease in piglets. In this method, using appropriate nutrition as fermentation medium, we enhanced transformation of active components in traditional Chinese medicine through activities of microorganism growth and metabolisim to change property reduce toxic and side effects and boost effectiveness of the medicine.To research the effect of Chinese Herbal Medicine microorganism fermentation products on prevention and treatment of ETEC diarrhea, in this study, firstly, culture medium and temperature, time, inoculation conditions of fermentation which Lactobacillus rhamnosus ATCC53103 required were optimized. Then, selecting 20 kinds of traditional Chinese medicines, through the test of minimum inhibitory concentration and bacteriostasis in vitro, we found that radix scutelalariae had the best bacteriostasis effect on ETEC was established. Finally, the radix scutelalariae and Lactobacillus rhamnosus were fermented, and the fermentated products were tested by in vitro antibacterial test, polysaccharide content and animal test. The model of piglet diarrhea caused by ETEC was established. The same batch of 30-days piglets with the same growth condition and boday weight were divided into five groups A, B,C, D, E. Each piglet, except for group E, was fed 5m L Escherichia coli liquid with the concentration about 3.65×1010CFU/m L. When all piglets were diarrhea, A, B, C, D,group were fed with saline, fermentation liquid, enrofloxacin and radixscutellariae liquid. Their body weight, feed intake and other indexes were measured every two days. In the morning of day 7 and day 14, blood collected with an empty stomach, To measure then immune index. After 14 days, pathological section testing was conducted after the piglets were slaughtered and intestines were removed.(1) Through improving carbon and hydrogen sources, culture medium for thegrowth of lactobacillus rhamnosus ATCC53103 were 1% glucose, 3% peptone, 2%yeast powder, 0.5% sodium acetate anhydrous, 0.2% ammonium citrate, 0.2%dipotassium hydrogen phosphate, 0.02% Mg SO4·7H2O, 0.05% Mn SO4·H2O, and0.1% polysorbate-80. Improved culture medium has an obvious influence on LGG ATCC53103 and the number of bacterium reached to 6.8×109cfu/m L. The optimum fermentation conditions were as follows: 150 m L culture liquid were loaded into the250 m L range of the cone, p H modulation was 6, 4% bacteria inoculation, and temperature, speed, time were 37 ℃, 80r/min, 24 h.In Chinese Herbal Medicine screening experiment, through the determination of minimum inhibitory concentration of 20 kinds of Chinese Herbal Medicine, it is concluded that bacteriostasis concentration of honeysuckle, radix scutellariae, cortex phellodendri and coptis are equal and minimum. Then, measure the four traditional Chinese medicines were measured by bacteriotasis experiment to obtain that honeysuckle bacteriotasis ring(1.803±0.064cm), cortex phellodendri(1.640±0.624cm), radix scutellariae(2.057±0.070cm) and coptis(1.780±0.036cm).The results proved that the bacteriostasis of radix scutellariae was best.(2)Through the experiment of fermentation of radix scutellariae and Lactobacillus rhamnosus ATCC53103, the optimal fermentation conditions obtained were temperature 37°C, initial PH 6, revolving speed 80r/min, inoculation quantity of bacteria 4%, content of Chinese Herbal Medicine 13%, time 30 h. In the previous treatment work of animal experiment, daily feed intake and average weight of piglets in each treatment group had no significant difference, but Ig A level in the serum varies in different groups. The Ig A level in treatment groups was higher than blank groups and comparison groups, and ferment products groups was highest, have no significant difference(P﹤0.01). But during the late period, daily feed intake and average weight have obvious improvement, no significant difference showing between treatment groups and comparison groups(P﹤0.05), Ig A level in the serum of each groups lower but no obvious difference, fermentation products groups. Diarrhea rate, fecal scoring, intestines lesion level in the fermentation products groups were lower than other groups. Serum bacteriostasis experiment results, blank group(0.663±0.070cm), enrofloxacin group(1.603±0.064cm), fermentation liquid group(1.850±0.060cm), radix scutellariae(1.580±0.036m). Bacteriostasis ring in fermentation liquid groups was higher than blank groups and other comparison groups,and had significant difference. Thus, I concluded that fermentation proucts of radixscutellariae Lactobacillus rhamnosus has an significant effect in the prevention and treatment of colibacillus diarrhea in piglets. |
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