| Chicken Coccidiosis is caused by one or more Eimeria parasitic on the chicken intestinal epithelial cells,which is protozoan disease,seriously harm to the development of the chicken industry.At present,the treatment of chicken coccidiosis is mainly dependent on chemical drugs,but the long-term use of chemical drugs causes drug-resistant strains and drug residues affecting food safety.Chinese medicine and probiotics compared to chemical drugs,have safety,environmental protection,enhancement of the body immunity,simple preparation process and many other advantages.Therefore,traditional Chinese medicine and probiotics fermentation technology has become a new direction in animal disease prevention and control.In order to explore the therapeutic effect of Chinese herbal medicine and probiotic fermentation products on chicken coccidiosis,this experiment first screening Chinese herbal medicine and traditional Chinese medicine prescriptions by inhibiting the oocysts of Eimeriatenella in vitro,and the traditional Chinese medicine prescription which in vitro inhibition of coccidia sporulation is remarkable was inoculated with Lactobacillus rhamnosus ATCC53103 to prepare the compound Chinese medicine fermentation preparation.Finally,the therapeutic effect on the chicks with infected coccidia oocysts was studied in order to evaluate the effect of the compound Chinese medicine fermentation on the treatment of chicken coccidiosis.The specific research contents and test results are as follows:1.Chinese medicine screening and compound Chinese medicine probiotics fermentation conditions to determine:First,the three herbivores of Artemisia annua,Sophoraflavescens and Bupleurum work best in inhibiting the sporulation of coccidia oocysts by in vitro inhibition of coccidial oocysts.with this three kinds of traditional Chinese medicine as the main drug anthelmintic composition of Chinese herbal compound,Again by the test in vitro inhibition of coccidia sporulation to screen compound traditional Chinese medicine,finally we get the best prescription on inhibition of oocyst sporulation: Sweet Wormwood Herb(20%),Changshan(20%),Pulsatilla(15%),Agrimonia pilosa Ledeb(15%),Sophora japonica.L(10%),Scutellaria Baicalensis(10%),Glycyrrhiza uralensis(10%).The conditions were optimized in the system using the above prescription and Lactobacillus rhamnosus ATCC53103 strain to co-ferment by the number of live bacteria as indicators,which are the amount of traditional Chinese medicine(TCM)adding,the fermentation time,strain inoculation amount,the fermentation temperature and the fermentation speed.Using the above conditions for fermentation,compound Chinese medicine fermentation formulations are preparated.Analysis of changes before and after fermentation is performed byusing high performance liquid chromatography.The results showed that the optimum fermentation conditions were that the amount of traditional Chinese medicine was 100%,the inoculation amount was 4%,the rotation speed was 80 r / min,and above 12 h under 37?.The results showed that the fermentation of compound Chinese herbal medicine compared with that before the fermentation,its material composition changes indeed,and have the phenomenon of the rise and fall of different level.It indicates that ATCC53103 could make some ingredients in Chinese medicine change in the process of fermentation on Compound traditional Chinese medicine(TCM).Finally,through the acute toxicity test of chicks found that chicks in the daily equivalent of 48 g of the original drug containing traditional Chinese medicine fermentation were continuously fed 7 days,a total of the equivalent of 336 g of the original drug containing traditional Chinese medicine fermentation preparation.After the observation of the chicks and autopsy,we did not see any exceptions can prove that the compound Chinese medicine preparation fermentation is safe.2.The effect of compound Chinese medicine preparation and Lactobacillus rhamnosus ATCC53103 collaborative fermentation products on the treatment of chickens infected by E.tenella :Thirty 1 day-old chicks were randomly divided into 6 groups: A,B,C,D,E and F,with 5 in each one group.With the exception ofthe F group(blank control group),All chicks in the other A,B,C,D,E groups were inoculated with 5×104sporulating coccidia oocysts of Eimeriatenella at 13 days.The coccidial oocysts were detected in the feces of the chicks in each group from the date of the attack,and the coccidia oocysts were detectable in the feces of the all chicks at 18 days.Some chicks have hematochezia phenomenon,can determine the success of a chicken coccidian infection and the emergence of clinical symptoms.According to the following method for clinical treatment of determining coccidian infection in 18 days,A group were gavage each day fermentation preparation of compound Chinese Medicine 200 ?L,Group B were fed with 200?L of compound Chinese medicine,Group C was given 200 ?L(1 × 106 cfu / ?L)LGG ATCC53103,Group E were fed with 2‰ diclazuril by drinking water.After 5 days continuous administration,we observed clinical symptoms of installment chicks.The relative weight gain rate,cecal lesion score,cecal pathology,and per gram of feces oocyst value were measured in each group and the anti-coccidiosis index of each group was calculated.At the same time,the changes of CD4+and CD8+T lymphocytes in peripheral blood of chickens were detected by flow cytometry.The changes of IgG and SIgA contents in chicks and the expression of IFN-? and IL-2 were detected by ELISA,and the contents of nitric oxide and nitric oxide synthase in serum of chicks were detected.The results showed that the weight of chicks infected with coccidia oocysts increasedsignificantly after taking the compound Chinese medicine fermentation preparation,which was significantly different from that of the control group(P <0.05).In addition,the compound Chinese medicine fermentation preparation can significantly reduce the cecal pathological changes,and can reduce the chicks out of the infected chickens discharge.The ACI of the compound Chinese medicine fermentation preparation was 130.59,which reached the level of moderate anti-coccidia.The results of flow cytometry showed that the contents of CD4+and CD8+T lymphocytes in peripheral blood of chicks fed with compound Chinese medicine were increased.In addition,taking compound Chinese medicine fermentation preparation can improve the chicken IgG and SIgA content as well as IFN-? and IL-2 expression level.The detection of nitric oxide and nitric oxide synthase in serum showed that the compound Chinese medicine fermentation preparation could decrease the contents of nitric oxide and nitric oxide synthase after coccidia infection.Through the above results,the compound Chinese herbal preparation and Lactobacillus rhamnosus ATCC53103 co-fermentation products have significant therapeutic effect on chickens infected E.tenella,which not only can increase the weight gain of chicks,reduce cecal lesions,reduce the discharge of thiosis,reduce nitric oxide and monoxide Nitrogen synthase content,but also by excitatory SIgA antibody secretion,increased IL-2 expression levels,and enhance immunity.This study provides a theoretical basis for the prevention and treatment of chicken coccidiosis by traditional Chinese medicine fermentation technology using lactic acid bacteria. |
PDF Full Text Request