| Avian trichomoniasis is a serious infectious disease in brids like pigeon,chicken and engle,infection rate is very high.Trichomonas gallinae(T.gallinae)infection affects the production performance of adult pigeons seriously and even cause the death of squab.At present,traditional Chinese medicines and antibiotics are often used to cure this disease,but the cost of traditional Chinese medicines is high and antibiotics can make drug residue worsen.There is a need to screen a break by resistance breeding.Many previous studies focused on morphology,epidemiology,diagnosis and treatment of T.gallinae.There were few researches which focused on pathological damages and the different expressed genes in different tolerance individuals.Therefore,the aim of this research is to select resistance genes by RNA-seq through T.gallinae challenging,and investigate the function of exosomes protein.This research can lay a foundation to healthy production of pigeon industry.Prevalence of T.gallinae was investigated to determine the breed used in challenge test.T.gallinae was isolated from upper digestive tract of infected pigeons.T.gallinae was identified by microscopy and sequencing.After challenge nasally,susceptible group(S)and tolerance group(T)was distinguished by the number of T.gallinae in oral cavity.Pathological manifestations of individuals were investigated.Differentially expressed genes were identified by RNA-seq.Exosomes of T.gallinae were extracted by ultracentrifugation,and label-free was used to identify the proteins of exosomes.(1)Average infection rate of males was significantly higher than that of females(P < 0.05).Infection rate of adult pigeons was significantly lower than squabs(P < 0.05).Prevalence of T.gallinae in White King pigeon was the highest among the five breeds(P < 0.05).The parasite had T.gallinae morphology,and sequence alignment of ITS1/5.8S/ITS2 gene showed an identity of 98% which can be used in challeng test.(2)Apoptosis rates in tolerance group of crop and esophageal were significantly higher than other two groups(P < 0.05),while in spleen,apoptosis rate in susceptible group was higher than other two groups(P < 0.05).RT-PCR results of immune related factors showed that tolerance group had higher IL-1β and lower TGF-β in crop,esophageal and spleen.Expression of apoptosis related factors in susceptible group of spleen was higher,and expression of apoptosis related factors in susceptible group of liver was higher just at 1 DPI.(3)285 mRNAs and 890 lncRNAs were differentially expressed in C vs S,and 157 mRNAs and 828 lncRNAs were differentially expressed in C vs T,95 mRNAs and 771 lncRNAs were differentially expressed in S vs T.FAAH,DDIT4 and mTOR signaling may participate in the regulation of resistance to T.gallinae;DHODH,OTUD1 and Notch signaling may play a key role in the process of T.gallinae infection.(4)The isolated vesicles presented a typical cup-shaped morphology;NTA results showed that particle size concentrated on 125.1 nm;proteins such as CD63 and TSG101 were conspicuously detected,suggesting that the extracts were exosomes.GO and KEGG analysis showed that proteins of exosomes might play roles in energy metabolism,signal transduction and biosynthesis.In conclusion,compared to susceptible group,tolerance group may show the stronger immune response to T.gallinae infection,which can accelerate the elimination of parasites.Gene FAAH and DDIT4 may be candidate resistance genes to T.gallinae.And T.gallinae can secrete exosomes,proteins of exosomes might play roles in energy metabolism,signal transduction and biosynthesis. |
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