Effect Of Factors Challenge On The Immune Response Of Hemocytes In Scallop Chlamys Farreri

In this dissertation, hemocyte monoclonal antibodies were used to detect thevatiations of total hemocyte and granuloyte counts after the challenge of acute watertemperatures, air exposure, reproduction stress and Vibrio anguillarum, as well as therelationship of expressions of immune related genes with the water temperaturesafter Vibrio anguillarum challenge, aiming at accumulate data for studying effects ofthe aquaculture environment、 physiological conditions and pathogen challenge onthe immne defense of scallop. This dissertation includes the following two parts:1Effects of four factors on variations of hemocyte in scallop Chlamys farreriIn this chaper, scallop Chlamys farreri was disposed by challenges of acute watertemperature (transferring scallops from17℃to11℃,23℃and28℃)，air exposureat different temperatures (5℃,11℃,17℃and23℃), starvation, spawning andspermiation respectively, and then detected hemocyte count, granulocyte count andratio by enzyme-linked immunoadsorbent assay (ELISA) using monoclonalantibodies against hemocyte or granulocyte as the primary antibody separately andflow cytometry respectively. The results showed that: total hemocyte count (THC)increased significantly at11℃,23℃, and then recovered to the control level, whiledeclined apparently at28℃; granulocyte count (GC) and ratio (GR) weresignificantly higher at11℃, however at23℃,28℃, declined gradually to theminimum at24h, and at28℃GC and GR were always under the control levels.THC, GC and GR descended significantly after air exposure at5℃,11℃,23℃，andthe scallops challenged at5℃had a higher level when recovered in the seawater;there was no scallops survived after6h air exposure at23℃. The THC、GC and GRdropped challenged with starvation、 spawn or spermination and then recovered. Inconclusion: variations of hemocyte and granulocyte are sensitive to temperature challenge, lower at higher temperature28℃; air exposure has significant influence onhemocyte and granulocyte counts, with lowest survival rate at23℃. Starvation andreproduction stress leads to the decrease of THC、GC and GR.2Variations of hemocyte count and immune related genes expression of scallopChlamys farreri after Vibrio anguillarum challengeIn this paper, scallop Chlamys farreri was challenged with10~4、10~7CFU/mlVibrio anguillarum and PBS at different water temperatures11℃,17℃,23℃and28℃respectively, and then detected THC and GC by ELISA using monoclonalantibodies against hemocyte or granulocyte as the primary antibody separately,besides, detected temporal mRNA expressions of six immune related genes (LGBP,Lectin-2,Toll like receptor, Lysozyme, SOD and PO) by qRT-PCR. The resultsshowed that,10~4CFU/ml. THC and GC almost stayed at the control levels challengedwith10~4CFU/ml V. anguillarum；challenged with107CFU/ml, GC declined at28℃，while increased first and then recoveried at other temperatures, with higher at23℃；challenged with10~7CFU/ml, GC decreased at all temperatures, and declined higher athigher temperate. The expression of six immune related genes was higher at10~7CFU/ml than10~4CFU/ml group at same temperature; at higher temperatures,17℃、23℃, the expressions of genes were higher; while at28℃with higher bacteriachallenge, the expressions had a suppression at the later of the experiment. Inconclusion, challenged the Vibrio anguillarum, the variation of hemocyte and andexpression of immune related genes witnessed relationshiop with temperatures, moreobvious at higher temperature while at too higher temperature28℃，the immuneresponse of hemocyte was suppressed.