| Rabbit coccidiosis is a parasitic protozoa caused by coccidiosis of Eimeraceae.Among them,Eimeria skrjabini has the strongest pathogenicity and the greatest harm.At present,there is no effective vaccine to control rabbit coccidiosis,and chemical drugs are commonly used to control rabbit coccidiosis,but with the increasingly prominent problem of drug residues,there are more and more studies on the application of traditional Chinese medicine to control rabbit coccidiosis in recent years.Traditional Chinese medicine has a good development prospect in the treatment of Eimeria coccidiosis.In view of this,in order to find a compound prescription of traditional Chinese medicine with better therapeutic effect,25 kinds of traditional Chinese medicine with the main functions of antimicrobial and insect repellent,heat-clearing and detoxification,hemostasis and dysentery,protecting liver and gallbladder were selected in this study.the Chinese herbs with better effect were screened out through the experiment of inhibiting oocyst sporulation in vitro,which were composed of three groups of compound prescriptions,which were tested by anti-coccidiosis test in vivo and inhibition of oocyst sporulation in vitro.To analyze the anti-Eimeria coccidiosis effect of three groups of compound prescription,and get the traditional Chinese medicine compound preparation with better therapeutic effect,which provides a basis for the prevention and treatment of rabbit Eimeria coccidiosis and provides a reference for clinical application.The contents and results of the study are as follows:Experiment 1,Isolation,identification and purification of rabbit Eimeria.In August 2019,the livers of rabbits died of rabbit coccidiosis were collected from a rabbit farm in Maojiagou,Jingyue District,Changchun City,Jilin Province.After grinding and making 3x104 / m L sporulated oocyst suspension,Eimeria skrjabini oocysts were purified by centrifugation precipitation and saturated sucrose floatation.The size of oocysts was about 21.4 μ m × 19.3 μ m under microscope.The DNA of purified Eimeria skrjabini was extracted,isolated and purified.Nest PCR detection was used to further identify coccidiosis.The results showed that the size of the bands amplified by 18 Sr DNA genome and 23 Sr DNA genome was the same as that of the target band,and the sequencing results were 100% and 99% in NCBI BLAST,respectively.Experiment 2,Screening and prescription of single traditional Chinese medicine against Eimeria skrjabini.Search the literature combined with the clinical symptoms and pathogenic mechanism of E.stiedai to select a total of 25 traditional Chinese medicines with the main functions of anti-bacterial deworming,heat-clearing and detoxification,hemostasis and dysentery,protecting liver and gallbladder.Each traditional Chinese medicine was made into four concentrations of water extract of 0.2g/m L,0.15g/m L,0.1g/m L and 0.05g/m L,and mixed with unsporulated Eimeria oocysts.The oocysts were cultured in a constant temperature water bath at 28.2 ℃.The sporulation rate of oocysts was determined at 12 h,24h,36 h and 48 h,respectively.Experiment 3,Study on the effect of compound preparation of traditional Chinese Medicine in the treatment of Eimeria skrjabini.Three groups of prescriptions were made into four different concentrations of aqueous extracts of 1g/m L,0.7g/m L,0.5g/m L and 0.3g/m L,and mixed with unsporulated E.stiedai oocysts.The sporulation rate of oocysts was measured at 12 h,24 h,36 h and48 h,respectively.80 rabbits with roughly the same body weight were divided into three groups:diclazuril group,chlorophenylguanidine hydrochloride group,worm attack group,blank group,prescription group 1,prescription group 2 and prescription group 3.In vivo anti-coccidial test,liver lesion score,OPG value,oocyst value,survival rate,weight gain rate,liver weight index,blood routine index,blood coagulation four indexes,liver biochemical index.The results showed that the sporulation rate of three groups of prescriptions was the lowest when the concentration was 1g/m L,and the lowest sporulation rate of oocysts in three groups of prescriptions was only 37.17%,indicating that the inhibitory effect of prescription three on oocyst sporulation was higher than that of prescription one and prescription two.The relative weight gain rate of the three groups was84.65%,which was significantly different from that of the insect attack group,and the survival rate was 100%.The average score of liver lesion was 0.48,which was lower than that of the other groups and higher than that of the chlorophenyl guanidine hydrochloride group,and the OPG28 was0.14 × 104 g per day,which was only higher than that of the chlorophenyl guanidine hydrochloride group.The results showed that the three groups of prescription could improve the decrease of weight gain caused by Eimeria skrjabini infection in rabbits,improve the liver damage caused by Eimeria skrjabini oocysts,and reduce the number of oocysts in the cecum.28 days after rabbit infection with Eimeria skrjabini,the liver function of all the experimental groups except the blank group was damaged to varying degrees,among which the liver function of the rabbits in the challenge group was the most severe.Compared with other groups,the difference was extremely significant(P < 0.01).The clotting time was prolonged and the inflammation increased in vivo.The anti-coccidial effect of the three prescription groups was better than that of the first prescription group,the second prescription group and the diclazuril group,and slightly lower than that of the chlorpheniramine hydrochloride group.can effectively reduce the degree of liver function damage,shorten clotting time,reduce inflammation in vivo. |
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