| Objective:Coccidiosis is one of the most serious diseases endangering the poultry industry at present,which seriously limits the development of the poultry industry.The control of coccidiosis is mainly based on the addition of coccidiosis resistant chemical drugs and antibiotics in feed.However,the problems of drug resistance and drug residues in animal products have led scholars to find new effective and safer alternatives.Traditional Chinese medicine has a long history in China and is rich in traditional Chinese medicine resources.The development of traditional Chinese medicine has always been a research hotspot in China.Taking Xinjiang yellow feather broilers as the research object,this thesis aims to explore the effect of anti coccidiosis traditional Chinese medicine on the distribution and quantity of intestinal glial cells(EGCS)and the expression of Cx43 and iNOS in chicken intestine after bloody stool caused by E.tenella,in order to provide reference basis for the prevention and treatment of chicken coccidiosis.Methods:1.Establishment of artificial infection in Broiler E.tenella model was constructed with1×10~4,2.5×10~4,5×10~4and 7.5×10~4E.tenella sporulated oocyst was administered to 14-day-old chicks,and the clinical performance,stool score and pathological changes were recorded to determine the appropriate dose of infection.2.When the chicks were raised to the age of 12 days,their feces were checked to observe whether there were coccidial oocysts in the feces.If there were no coccidial oocysts,100 healthy chicks with similar weight were selected and randomly divided into 5 groups.At the age of 14 days,except the negative control group,the other 4 groups were orally administered with 2.5×10~4of normal saline containing sporulated oocysts was used to observe the health status of chickens every day after the eggs were administrated.When the chickens had symptoms such as bloody stool,anorexia,anemia and pallor of crowns and visible mucous membranes,the medication treatment was started.Test group A is the infection group,containing 2.5×10~4of normal saline for sporulated oocysts;Test B was the control group without infection and treatment;Test group C is group I of traditional Chinese medicine,containing 2.5×10~4sporulated oocysts were treated with 1 ml of normal saline and Chinese medicine I was mixed in the feed at the dose of 5 g/kg;Test D was group II of traditional Chinese medicine,containing 2.5×10~4sporulated oocysts were treated with 1 ml of normal saline and Chinese Medicine II was mixed in the feed at the dose of 5 g/kg;Test E is sq group,containing 2.5×10~4of normal saline of sporulated oocysts was mixed in water according to the dosage of Sulfaquinoxaline sodium.All groups were free to drink high-temperature sterilized water.After infection,the health status of chickens was observed every day.Five chickens in each group were slaughtered on the 8th day.Intestinal tissue samples were taken and stored in Helly fixed solution,10%neutral formaldehyde fixed solution and-80℃refrigerator for improved Holzer staining,immunohistochemical staining and ELISA detection.Result:1.Set up the inoculation dose of E.tenella infection model was 2.5×10~4sporulated oocysts/pigs.2.The results of improved Holzer staining showed that EGCs in the control group were distributed widely in the mucosa,submucosa and myometrium of each segment of the intestine,while the outer membrane was distributed sporadically and clustered in the lamina propria near the top of Villi,the rest of the body is scattered.There was no significant difference in the number of EGCs between the infected group and the control group(P>0.05).The main distribution of EGCs was in the cecum.The EGCs in the control group were more in the villus than in the duodenum,there were more in the lamina propria,muscularis mucosa,around the intestinal gland,submucosa and muscularis.Compared with the control group,the distribution of EGCs in the infected group decreased obviously,but there were few intestinal glands and only a few muscle layers.The number of EGCs in Caecum of the infected group was significantly lower than that of the control group(P<0.05),and the number of EGCs in intestinal segments of the SQ group was significantly lower than that of the control group(P<0.05),compared with the infected group,the number of EGCs increased significantly(P<0.05),but there was no significant difference between the two groups(P>0.05).3.The results of Elisa showed that the content of Cx43 in Caecum of the infected group was significantly lower than that of the control group(P<0.05),there was no significant difference among the three groups(P>0.05).The content of CX43 in Caecum of the three groups was lower than that of the control group(P<0.05),but higher than that of the infection group(P<0.05).The inducible nitric oxide synthase(iNOS)in Caecum of infected group was significantly higher than that of control group(P<0.05).There was no significant difference among three groups(P>0.05),the content of iNOS in cecum of three groups was higher than that of control group(P<0.05),but lower than that of infection group(P<0.05).4.The results of immunohistochemistry showed that Cx43 was mainly expressed in the mucosal epithelium,intestinal gland,submucosa,lamina propria and myenteric plexus of the control group,and the expression of Cx43 in small intestine of each experimental group was similar to the control group,however,the expression of Cx43 in caecum of the infected group was significantly lower than that of the control group(P<0.01),and there was no significant difference in the expression of Cx43 in caecum among the three groups(P>0.05),Cx43 in cecum of 3 groups was significantly lower than that of control group(P<0.05),and significantly higher than that of infection group(P<0.05).iNOS was weakly expressed in control group,and INOS was weakly expressed in small intestine in infection group,Chinese Herbal Medicine Group I,Chinese Herbal Medicine Group II and SQ Group.In caecum,iNOS was mainly expressed in intestinal gland,mucosa epithelium and villus lamina propria,and the expression was significantly increased in caecum(P<0.05)The expression of iNOS in caecum of Group I,Group II and group SQ had no significant difference(P>0.05).The expression of iNOS in caecum of three groups was significantly higher than that of control group(P<0.05),compared with the infection group,the difference was significant(P<0.05).Conclusion:Set up the inoculation dose of E.tenella was 2.5×10~4sporulated oocysts/chickens,and the infected chickens were infected with E.tenella,the distribution of EGCs in Caecum was changed and the number of EGCs was decreased,which destroyed the EGCs network barrier of Caecum,and the traditional Chinese medicine could promote the secretion of CX43 in caecum,decrease the secretion of iNOS in caecum and inhibit The pathogenicity of E.tenella and its protective effect on intestinal mucosa... |
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