| The rodlet cell has been found for over 100 years in teleostean species,at present,the recent part of study indicate that RC may be a specialized cells associated with fish non-specific immune response.There has been debate on the nature,function and evolutionary status of the RCs by far,and the research data of molecular biology about this cell is almost a blank.The Yellow River carp?Cyprinus carpio?is utilized as test animal of this research,and the research project includes the following contents:?1?The criterion of the cells morphology are established by light microscope and electron microscope,the results show that: On the morphology,Rodlet cell has a thick fibrous capsule,and the rodlet is exist in cytoplasm,its appearance looks be rodlike,the core of the rodlet is high electron density;the developing of RC contains three stages: young stage,mature stage and discharge stage,Immature rodlet cells were those cells possessing a thin fibrous capsule and rodlets without dense cores,Mature cells were those with a well developed fibrous capsule,multiple rodlets possessing dense cores and a small basal nucleus containing electron dense deposits.The apex of the cell possessed numerous mitochondria,vacuoles and ribosomes.In the discharge stage,the cell can release the rodlet from the cell and the core of the rodlet.?2?This study take wound as the stimulating factors for yellow river carp,then draw materials at 0h,0.5h,1h,2h,4h,6h,12 h,24h,48 h,72h,96 h after wound,the materials contain forgut,trunk kidney,head kidney,gill and skin.Normal anatomy and histology method is applied to study the distribution and morphology observation in organizations.The results show that forgut,RC can be seen in trunk kidney,head kidney and gill,except in skin.In statistics,Compared with control group,the number of the RC tend to rise before falling along with time,in forgut,the number of RC rise to peak after 1h,the distribution proportion has been 7.6431×10-5 individual /?m2;in trunk kidney,the number of RC rise to peak after 2h,the cell density has been 7.7652×10-5 individual /?m2;in head kidney,the number of RC has two peaks,the first peak appeared after wound 4h,the cell density has been 5.6819×10-5 individual /?m2,the second peak appeared after wound 12 h,the cell density has been 7.1549×10-5 individual /?m2.?3?As2O3 exposed to the yellow river carp to observe the distribution and variation trend of RC in tissues,the preliminary experiment set up 8 concentration treatment groups,they respectively were 0?10?20?30?40?50?60?70?80?90 mg/L,after exposed 96 h,record the death rate to get the limiting concentration respectively be 20 mg/L and 70 mg/L,then according to the improved karber method,set up 7 concentration treatment groups,they respectively were 0?20?25.7?33?42.4?54.5?70mg/L.Record the death rate to get the LC50?96 h?and SC?96 h?.and draw materials: forgut,truank kidney,head kidney,gill,liver and skin,then observition and statistic the date by conventional anatomy and histology.The results showed that: At temperature 25 ± 0.5 ?,p H 7.0 ± 0.1,As2O3 was highly toxic substance on the Yellow River carp.The median lethal concentration of 96 h was 30.3 mg/L,The 96 h safe concentration value was 3.03mg/L.RC can be seen in forgut,trunk kidney,head kidney,gill and liver;in statistics,According to the statistical analysis,in the acute exposure test,the different concentration of As2O3 exposed yellow river carp for 96 h,the results show that the number of RC tend to rise before falling along with the rising of concentration,the peak is concentrated near the median lethal concentration of 96 h,in forgut,the number peak is respectively at 26mg/L and 42 mg/L,the cell density is respectively 4.9327×10-5 individual/?m2 and 5.2645×10-5 individual /?m2;in trunk kidney and head kidney,the number peak is at 33 mg/L,the cell density of them are respectively 1.5046×10-4 individual /?m2 and 8.4176×10-5 individual /?m2.In the subacute exposure test,we take the 96 h safe concentration value as experiment concentration,and draw materials at 0h,6h,12 h,24h,48 h,96h and 7d to observe the change on number of RC.The results show that the number of the RC tend to rise before falling along with time,in forgut,the number of RC reached the peak at after exposed 96 h,the cell density has been 6.8051×10-5 individual /?m2;in trunk kidney and head kidney,the number of RC reached peak at after exposed 24 h,the cell density has respectively been 1.3819×10-4 individual /?m2 and 1.1152×10-4 individual /?m2.?4?Aeromonas hydrophilia infect the yellow river carp,then observe the distribution and variation trend of RC in tissues,the preliminary experiment set up 8 concentration treatment groups,they respectively were contral?0.7%Nacl?,1×105,5×105,1×106,5×106,1×107,5×107,1×108 CFU/ml.At 96 h After infection,record the death rate to get the limiting concentration respectively be 5×105 CFU/ml and 1×108 CFU/ml,then according to the improved karber method,set up 7 concentration treatment groups,they respectively were contral?0.7%Nacl?,5×106,9.1×106,1.66×107,3.02×107,5.5×107,1×108CFU/ml.Record the death rate to get the LC50?96 h?and SC?96 h?.The results show that: At temperature 25 ± 0.5 ?,p H 7.0 ± 0.1,The median lethal concentration of 96 h was 4.98×105 CFU/ml,The 96 h safe concentration value 4.98×104 CFU.ml-1.in the subacute toxicity test,we take the 96 h safe concentration value was experiment concentration,and draw materials at 0h,6h,12 h,24h,48 h,96h,7d and 15 d to observe the change on number of RC,RC can be seen in forgut,trunk kidney,head kidney,gill and liver;in statistics,the number of RC have two peaks in frogut and head kidney,respectively at after infection 12 h and 96 h,the cell density at 12 h were respectively 1.9238×10-4 individual /?m2 and 5.2008×10-5 individual /?m2,the cell density at 96 h were respectively 1.9238×10-4 individual /?m2 and 5.2008×10-5 individual /?m2.In trunk kidney,the number of RC reached peak at after infection 24 h,the cell density was 1.2532×10-4 individual /?m2?Through the results of experiment above,this paper can get the following conclusions:?1?The criterion of the cells morphology are established by light microscope and electron microscope,and RC is the endogenous cell in the yellow river carp;?2?Three kinds of the experimental results show that the different stressors can lead to the RC of Yellow River carp proliferation in the body,which is disproving the role of the RC is resist stress;?3?select one kind of stress process that can significantly increased the RCs number in fish tissue,and select the fish organ or tissue in which there is high abundance of cell though three experimental results,it's forgut of yellow river carp after infection Aeromonas hydrophilia 96 h.Lay the foundation of the separation of the RC. |
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