海豚链球菌3种免疫原对牙鲆血细胞吞噬及SOD、POD和ACP活力的影响

本文制备海豚链球菌(Streptococcus iniae)强毒株NUF849的福尔马林灭活全菌(FKC)、胞外产物(ECPs)以及全菌与胞外产物的混合物(FKC+ ECPs)3种免疫原,以其免疫牙鲆(Paralichthys olivaceus),并在免疫牙鲆后第42天分别以海豚链球菌NUF849和NUF812攻毒.在免疫牙鲆后第7、14、21、28、35、42天及攻毒后第7天分别取样,测定血细胞吞噬能力及血清中超氧化物歧化酶(SOD)、过氧化物酶(POD)和酸性磷酸酶(ACP)的活力,以比较3种免疫原对这些免疫指标的影响.结果显示,免疫后尤其攻毒后,红细胞、淋巴细胞、单核细胞、嗜中性粒细胞等的吞噬率与吞噬指数均上升,FKC+ ECPs组显著高于其他组(P＜0.05);各免疫组SOD活力随时间延长而下降并趋于稳定,攻毒后各组数值有不同程度的升高,但FKC+ ECPs组数值最低;免疫组POD活力先升高,第28天起下降,攻毒后各组均下降,FKC+ ECPs组始终高于其他免疫组(P＜0.05);免疫组ACP活力也在免疫后第28天达到峰值,以某株NUF849攻毒后FKC+ ECPs组水平上升(相对于免疫后第42天),其他组显著下降;FKC、ECPs、FKC+ ECPs对菌株NUF849的相对免疫保护宰分别为50％、40％和90％,对NUF812的相对免疫保护率分别为60％、30％和80％,FKC+ ECPs组显著高于其他组(P＜0.05).以上研究表明,各免疫组均诱导了牙鲆的非特异性免疫应答,具有一定的免疫保护效果,其中FKC+ECPs组效果最佳.     

Anti-enrofloxacin antibody production by using enrofloxacin-screened HSA as an immunogen

A two-step zero-length cross-linking procedure using active esters was successfully adopted for conjugating enrofloxacin （EF） to human serum albumin （HSA）. The derived conjugate was characterized by UV spectrum and then used for immunization of BALB/C mice. In enzyme-linked immunosorbent assay （ELISA） and competitive inhibition ELISA experiments, the derived antiserum exhibited high antibody titer （greater than 1 ： 250 000） as well as varied cross-reactivity （from 97.8% to 161.7%） to three analogs of EF belonging to fluoroquinolones family. But over the concentration range studied, no significant cross-reactivity was observed to other group of antibiotics （chloramphenicol, oxytetracycline, sulphamethoxazole and nysfungin）. It was confirmed that the synthesized immunogen was highly antigenic and elicited specific antibody responses in BALB/C mice against EF.