Effects Of Decoquinate And Artemisinin On Chickens Coccidiosis Infected By Eimeria Tenella

ObjectiveOur previous research shows that Artemisinin(ART)and its original plant Artemisia annua Linn(AAL)have certain anticoccidial effects,which can significantly reduce the clinical symptoms of coccidiosis infected with Eimeria tenella(E.tenella)by promoting the apoptosis of infected cells,reducing tissue inflammation,and inhibiting the formation of oocyst wall.Based on this,we focused on evaluating the anticoccidial effect of ART and its derivatives,AAL compound preparation and ART combining decoquinate(DQ)in this study.In order to obtain the potential mechanism of DQ anticoccidial action,the iTRAQ technique was applied to detect the differential expression of the chickens cecum tissue proteins of the infection group and the DQ treatment group.MethodWe established the chicken coccidiosis model by artificial infection of 50,000 E.tenella sporulated oocysts to 15-day-old chickens.The experimental chickens was treated 24 h before inoculating the oocysts.The administration of medicine stopped on the seven days after inoculation.We dissected the chickens on the 180 h after the inoculation.In order to compare the preventive effects of drug treatment group of E.tenella in broilers,the relative weight gain rate,relative survival rate,lesion score of cecum,number of oocyst and ACI were recorded and used as the criterion.We took cecal tissue for sectioning to observe the appearance and pathological changes and observed the changes of the ultrastructure of the intestinal mucosa by field emission scanning electron microscopy.The cecal tissue of experimental chickens was collected for cryopreservation.After protein quantification,SDS-PAGE electrophoresis was applied to detect protein quality.LC-MS/MS and HPLC fractionation were performed after protein tryptic digestion and addition of 8 plex.We analyzed the differential proteins by the data obtained.Results1.Among ART and its derivatives,only ART can significantly reduce cecumlesions and reduce oocyst volume in infected chickens,with the ACI of 158.47.While ARM and ARS show no significant anticoccidial activity.2.Compared with the EI group,the AAL+dichroae radix group and the AAL+dichroae radix+bidens pilosa group significantly reduced the oocyst value and promoted weight gain,and improved cecal lesions.Their ACI were 153.34 and 147.32.The AAL+dichroae radix+bidens pilosa+pulsatilla had the best preventive effect among the three traditional Chinese herbs compound recipe groups.Its the relative weight gain rate is 90.08%and its ACI reached 161.7.Although it did not achieve the preventive effect of sulfamonomethoxine(the ACI was 184.6),it could significantly ameliorate cecal lesions and increase weight gain.3.The results showed that bloody feces and oocysts did not appear in the ART + DQ60 group.Its ACI was 196.22.A small amount of bloody feces and oocysts were present in DQ60 and ART+DQ40 groups,and their ACI were 187.15 and 185.72.ART + DQ20 group and ART group produced a large amount of bloody feces and oocysts.Microscopic lesions were observed in the cecum structure of ART + DQ60 group and ART + DQ40 group was better than those of DQ60 group and ART group.Observed from the arrangement,length,and crypt appearance of cecum microvilli,the microscopic structure of ART + DQ40 group was superior to that of DQ60 group.The study suggested that the combinations of ART with DQ40 and DQ60 have a synergistic effect in the control of E.tenella infection.4.The analysis identified 3410 proteins contained at least two unique peptides.The differential proteins were screened at a threshold of up-regulated or down-regulated 1.2 fold(FC=fold change,FC ? 1.2,P<0.05;FC? 0.83,P<0.05).Compared with the Blank Control group(BC),there were 155 significant differences in protein expression,370 expression levels decreased in the El group.According to the pathway enrichment method,compared with the BC group,five differential proteins in the EI group were enriched in the peroxisome proliferater activated receptor(PPAR)pathway.They were FATP(FC=0.18),L-FABP(FC=0.19),PEPCK(FC=0.71),CPT1A(FC=0.82),APOA1(FC=1.45).Compared to the IE group,there are 196 differential proteins annotated with GO function in the DQ group.The 96.6%of them had binding and 84.1%had catalytic activity.The proteins with significant differences of DQ group was enriched in the oxidative phosphorylation signal pathway,and there were 5 proteins:ATP7(FC=1.30),NDUFB4(FC=1.41),NDUFA8(FC=1.26),NDUFA10(FC=1.21),NDUFA8(FC=1.20).Compared with the BC group,there were 43 up-regulated proteins and 136 down-regulated proteins in the DQ group.This suggests that the pathogenesis of E.tenella infection in chickens may be related to the PPAR signaling pathway and DQ may play anticoccidial effects through oxidative phosphorylation signaling pathways.