Taxonomic Analysis And Detection Of A.ocellatum & C.Irritans Depending On RDNA Sequences

Amyloodinium ocellatum and Cryptocaryon irritans are common parasitic disease in the process of aquaculture that cultured Pseudosciaena crocea,which cause diseases outbreaks meanwhile will be highly detrimental to the economic income.Pseudosciaena crocea as a research object,determine an experimental method that we can obtain the mature trophont of Amyloodinium ocellatum.In the process of observing Amyloodinium ocellatum life history,we clarify life cycle process of Amyloodinium ocellatum on the Pseudosciaena crocea.in the meantime,determines the quantitative relation that the daughter cells number and the diameter of the tomont of Amyloodinium ocellatum and that the dinospore and the daughter cells number in the tomont.Furthermore,determine the life span of the dinospore.Grouping Pseudosciaena crocea in two groups,one specification is 72.14±16.64 and the other specification is 18.14±5.21g.we ascertain the minimum lethal dosage respectively.Determine the median lethal dose of the 18.14±5.21g Pseudosciaena crocea.estimate Amyloodinium ocellatum infection rate of 72.14±16.64g Pseudosciaena crocea.determines the toxicity of Amyloodinium ocellatum on the Pseudosciaena crocea preliminarily.Taxonomic status is still in doubt after two kinds of parasite taxonomic position changed many times,at the same time,two kinds of parasite disease control and prevention is still difficult to solve.This article cloned 18S/ITS 1/5.8S/ITS2/28S of Amyloodinium ocellatum.Analyse genetic relationship depending on the complete 18S and 28S sequence of Amyloodinium ocellatum.Cloned 18S/ITS1/5.8S/ITS2/28S/IGS of Cryptocaryon irritans.Analyse genetic relationship depending on the complete 18S and 28S sequence of Cryptocaryon irritans.At the same time,Analyse genetic relationship depend on IGS sequence of four Cryptocaryon irritans strains.In order to detect Amyloodinium ocellatum and Cryptocaryon irritans conveniently,this paper respectively design two pairs of primers in different genetic regions in the ribosome.The specific primers we design detect the dinospore and the trophont of Amyloodinium ocellatum in the ITS2 and 5'ETS.the sensitivity is 32 in the detection of the spores to 32 and is 1 in the detection of the between 20 and 45 microns trophont.It is hard to detect more than 60 microns ones.In the 28S sequence and IGS sequence of Cryptocaryon irritans,we design specific primers to detect the theront and the trophont of Cryptocaryon irritans.For the 2 pair of primers are highly sensitive to the theront and trophont.1 theront can be checked by PCR.One trophont of the different size(80-420 microns)can be checked by PCR.The sensitivity of the detection is the same between trophont and theront Two pairs of primers with species are highly species-specific and suitable for PCR primers.