保幼激素类似物对红火蚁的作用研究进展

我国发现红火蚁已有4年,但防治技术、产品依然匮乏。为了改善、提高我国红火蚁的防治技术水平,本文就保幼激素类似物(JHA)对红火蚁的作用表现和防治效果进行了总结分析。JHA可以造成蚁后卵巢萎缩、产卵量减少,导致发育畸形和蚁群等级比例失调,如工蚁数量减少、生殖型幼虫比例以及处女蚁后数量增加等,并最终导致整个蚁群死亡。JHA杀虫剂活性高、低残毒、对环境污染小,在田间防治红火蚁效果彻底,并可有效地防止防治区红火蚁种群的再入侵（re invasion）,但JHA毒饵也存在着防治效果缓慢等问题。本文就克服此类问题作了初步的探讨。     

吡虫啉对豌豆蚜的亚致死效应

室内测定了吡虫啉的亚致死剂量（LC20、LC30、LC40）对豌豆蚜的生长发育、繁殖和种群参数的影响。结果表明,随着吡虫啉亚致死剂量的增加,豌豆蚜F0代成蚜的寿命显著缩短,产蚜量显著降低。亚致死剂量也使F1代种群的内禀增长率、净增殖率和周限增长率降低,而发育历期、平均世代周期和种群加倍时间延长。     

Ozone Efficacy as a Bactericide in Seafood Processing

Abstract

The efficacy of ozonated water (0.6-1.5 ppm) was evaluated as a bactericidal agent for sanitizing food contact surfaces and for treatment of raw seafood. The presence of ozone reduced the bacterial levels substantially on stainless steel surfaces and to a lesser extent on plastic cutting boards. Ozone was about as effective as chlorine in lowering levels of Listeria innocua on inoculated food contact surfaces. Fish processing residuals present on the surface greatly reduced sanitizer effectiveness. In high organic conditions, chlorinated water was slightly more effective than ozonated water. However, ozonated water applied to fish fillets and roe was not effective for bacterial control. The presence of organic material, particularly with fillets, reduced the effectiveness of ozone. Ozone accelerated the development of rancidity in frozen roe and fillets, resulting in reduced shelf life. We recommend ozone only as a sanitizer for cleaned seafood contact surfaces.     

Incidence of Pathogenic Microorganisms in Aquacultured Rainbow Trout (Oncorhynchus mykiss)

ABSTRACT

Quantitative levels of six known pathogens (Listeria monocytogenes, Clostridium botulinum, Salmonella species, Vibrio cholerae, Yersinia enterocolitica, and Y. pseudotuberculosis) and aerobic plate counts were measured at five aquaculture facilities. The farmed rainbow trout (Oncorhynchus mykiss) and trout fillets were sampled at two different growing seasons to monitor for microbial hazards. Listeria spp. was identified in both whole trout and trout fillets from all five facilities sampled from both growing seasons. Presumptive Clostridium botulinum spores were also identified from all five facilities for both seasons. The growing season did not affect pathogen levels and there was no evidence that any one aquacultural system was superior to the others. Salmonella spp., Vibrio cholerae, and Yersinia spp. were not isolated from any of the trout samples analyzed.     

Effect of Trisodium Phosphate or Water Dip on the Survival of Salmonella and Listeria monocytogenes Inoculated Catfish Before and After Freezing

Salmonella and Listeria monocytogenes (LM) are occasional contaminants on raw fish. Catfish fillets were artificially contaminated with LM and Salmonella, dipped in a 1.5 % (30 min) trisodium phosphate solution (TSP), and cryogenically frozen. After 3 months frozen storage, Salmonella (2 log), but not LM, was inactivated on the fillets treated with the dip. This indicates that TSP dip followed by cryogenic freezing can be used to control Salmonella, but not LM, on catfish fillets.     

单核细胞增生性李斯特菌感染ICR小鼠模型的建立

单核细胞增生性李斯特菌(Listeria monocytogenes,LM)是一种人畜共患食源性致病菌,能引起人和动物较为严重的感染症状。现广泛使用具有免疫活性的小鼠模型测定半数致死量(LD₅₀)来评价不同LM菌株的致病性。为建立LM的ICR小鼠模型,本研究采用1/2a血清型的N21 LM菌株,分别以10⁶、10⁷、10⁸、10⁹和10¹⁰ CFU的剂量口腔灌注感染6周龄ICR小鼠,每组10只;另取10只接种PBS作为对照组,测定LM对ICR小鼠的LD₅₀;另取40只小鼠,平均分为2组,公母各半,以测定的LD₅₀剂量接种LM,分别对各组小鼠临床症状、组织病理变化、体重变化及组织中细菌载量进行评价。结果发现,N21 LM菌株对ICR小鼠的LD₅₀为10^9.25 CFU;感染周期为10 d左右,感染小鼠出现被毛粗糙、精神萎靡、阴茎垂出及体重下降等临床症状。组织病理学分析结果显示,肝脏先后出现实质内灶状细菌团块、形成血栓、细胞坏死等;脾脏主要表现为白髓内淋巴细胞减少;肺脏主要表现为纤维素性肺炎。菌落计数和Real-time PCR的检测结果发现肝脏中细菌载量最高,脾脏次之。结果表明,ICR小鼠能作为单核细胞增生性李斯特菌的感染模型。本试验结果为研究LM的致病机制、疫苗研究及抗菌肽转基因小鼠抗LM的评价奠定了基础。     

单核细胞增生李斯特菌 sRNA 伴侣分子 hfq的基因克隆、表达及纯化

为克隆、表达单核细胞增生李斯特菌sRNA分子伴侣蛋白hfq基因,并纯化重组蛋白,通过PCR的方法从单核细胞增生李斯特菌基因组DNA中扩增出hfq基因,将扩增产物克隆于pMD18-T载体中,测序验证后,再将hfq基因亚克隆至表达载体pET-32a（＋）中,成功构建pET-32a（＋）-hfq原核表达载体。然后,转化至大肠杆菌BL21（DE3）感受态细胞中,通过IPTG进行诱导表达,对表达蛋白进行可溶性分析,并采用Ni-NTA纯化目的蛋白。结果显示：克隆的hfq基因全长234 bp,编码77个氨基酸,通过SDS-PAGE可以检测到27 kDa的蛋白特异性条带;并从上清中纯化得到了Hfq融合蛋白。为进一步研究该蛋白的生物学功能奠定了基础。     

产单核细胞李斯特菌溶血素O单克隆抗体的制备与鉴定

【目的】制备抗产单核细胞李斯特菌溶血素O(LLO)的特异性单克隆抗体。【方法】用LLO重折叠的原核表达产物LLO-his免疫Balb/c小鼠,取其脾细胞,与骨髓瘤细胞在PEG4000作用下融合,经间接ELISA法筛选、多克隆及单克隆分离出阳性细胞株后,再用体内诱生腹水法大量制备单克隆抗体。用辛酸-硫酸铵沉淀法纯化单克隆抗体,间接ELISA法测定其效价及相对亲和常数;用HBT单克隆抗体亚类鉴定试剂盒鉴定抗体亚型;常规方法制备染色体,观察细胞株染色体数目及标志染色体;Western blot检测抗体的特异性。【结果】获得了2株稳定分泌抗LLO单克隆抗体的杂交瘤细胞系,分别命名为3F5-G3和2B4-C8,其腹水效价分别为1∶(1×105)和1∶(1×107);染色体分别为97±5和93±8条,且观察到标志染色体;相对亲和常数分别为0.49和0.26μg/mL;抗体亚类均为IgG1,轻链均为κ链。特异性鉴定结果表明,2种单克隆抗体均可与LLO蛋白发生特异性反应,而与载体蛋白不发生反应。【结论】获得了抗LLO的特异性单克隆抗体。     

猪伪狂犬病毒H株gE基因的克隆及序列分析

参照已发表的扩增伪狂犬病毒gI和gE部分基因的PCR方法,合成了一对引物,以PRV H株细胞毒为模板,扩增出一条约850 bp的片段,并进行克隆和测序。然后与GenBank上国内外其它毒株gE基因进行核苷酸和推导的氨基酸序列的同源性分析,构建了遗传进化关系图。结果表明,PRV H株与其他15株PRV核苷酸和氨基酸的同源性分别为 97.0%～99.2%和93.2%～98.5%,其中与SH株最近,分别为99.2% 和98.5%;进化树分析表明,该毒株与目前国内流行的毒株在同一进化分支内,与国外毒株分属不同分支,说明分离的H毒株与目前国内流行的毒株一致。     

9种药剂对斜纹夜蛾的室内毒力与田间应用效果

通过室内毒力测定和田间药效试验,测定当前9种主流杀虫剂产品对斜纹夜蛾的防治效果。结果表明,甲维盐、茚虫威、虫螨腈和虫酰肼对斜纹夜蛾具有优良的防效,可作为防治斜纹夜蛾的首选药剂。Bt与多杀菌素的效果次之。阿维菌素、高效氯氰菊酯和毒死蜱对斜纹夜蛾已逐渐失去控制作用,在生产中应尽量少用、慎用。