The Investigation Research Of Actinospores From Pond And RT-PCR Methond To Detect Myxobolus Honghuensis

There is a very high diversity of myxosporean in China, and some myxosporean species even have caused severe economic loss in aquaculture,and Jiangsu province is the main breeding area of crucian carp in China,it is usefull to make a suvery to study this kind of parasite. We investigated pond-reard crucian carp of Changzhou, Dafeng, Jiangdu city form May2013 to Sep2013. Myxosporean have two stages, the actinosporean stage and the myxosporean stage.In the present study, we investigated and studied of five kind of actinospore in crucian carp in Jiangsu region, and established a specific and sensitive RT-PCR test method for Myxobolus honghuensis, and established an artificial infection experimental, so as to provide some theoretical basis for life cycle of myxosporeans and prevention of myxosporidiosis.1. Investigation of actionspore types in pond breeding crucian carp and taxonomic study. Investigation of myxosporeans parasitic in crucian carp and taxonomic study. In this present study, five actinosprean types-Guyenotia type, Triactinomyxon type,Raabeia type.Aurantiactinomyxon type and Neoactinomyxum type were collected from the oligochaete Branchiura sowerbyi Beddard collected from crucian carp (Carassius auratus gibelio) ponds in Changzhou city and Jiangdu city, Jiangsu province.On the basis of morphology data, these five actinosprean types presented in this study differs from those already known in the literatures.Based on DNA sequences analysis, showed the Guyenotia type examined present 98.2% genetic similarity to Zschokkella sp.(DQ118776), Triactinomyxon type examined present 86.3% genetic similarity to Chloromyxum legeri(AY604197),Raabeia type examined present 98.8% and 98.5% genetic similarity to Myxobolus cultus (AB121146) and Myxobolus cultus (HQ613409),Aurantiactinomyxon type and Neoactinomyxum type examined present 95.9% and 93.8% genetic similarity to Thelohanellus wuhanensis (JQ968687), thus the author indicated that the type of Guyenotia actinospore considered as actinosporean stage of Zschokkella sp.,the Raabeia type considered as Myxobolus sp.,Aurantiactinomyxon type and Neoactinomyxum type considered as Thelohanellus sp..On the basis of morphology data, these five actinosprean types presented in this study differs from those already known in the literatures.2. A detecte method for Myxobolus honghuensis by RT-PCR. RT-PCR reactions were carried out using the universal eukaryoticprimers ERIB1 & 10 and the M. honghuensis specific primer HF-F&R.The RT-PCR showed that only M. Honghuensis has the amplification curve. The sensitivity of RT-PCR was showed that three Myxobolus honghuensis could be dected.3. The artificial infection B. sowerbyi experimental based on the myxosporeans on the throat which collected from infected fish.Then,spent nearly one month to detect and analyze the effect of infection after three month, found that b.sowerbyi didn’t release any type of actionspore after feeding myxosporean three months later.