食品中单增李斯特菌检测技术研究进展

单增李斯特菌是一种重要的人畜共患食源性致病菌,如何有效控制食品(尤其是即食食品)中的单增李斯特菌,是食品安全的重要任务,其中快速、准确的检测技术是关键因素之一.作者对单增李斯特菌的传统检测法、免疫学检测法和分子生物学检测法进行综述,并对目前国内单增李斯特菌检测过程中所存在的问题进行探讨,以期为今后的研究提供参考.     

Exendin-4 improves resistance to Listeria monocytogenes infection in diabetic db/db mice

The incidence of diabetes mellitus is increasing among companion animals. This disease has similar characteristics in both humans and animals. Diabetes is frequently identified as an independent risk factor for infections associated with increased mortality. In the present study, homozygous diabetic (db/db) mice were infected with Listeria (L.) monocytogenes and then treated with the anti-diabetic drug exendin-4, a glucagon-like peptide 1 analogue. In aged db/db mice, decreased CD11b⁺ macrophage populations with higher lipid content and lower phagocytic activity were observed. Exendin-4 lowered high lipid levels and enhanced phagocytosis in macrophages from db/db mice infected with L. monocytogenes. Exendin-4 also ameliorated obesity and hyperglycemia, and improved ex vivo bacteria clearance by macrophages in the animals. Liver histology examined during L. monocytogenes infection indicated that abscess formation was much milder in exendin-4-treated db/db mice than in the control animals. Moreover, mechanistic studies demonstrated that expression of ATP binding cassette transporter 1, a sterol transporter, was higher in macrophages isolated from the exendin-4-treated db/db mice. Overall, our results suggest that exendin-4 decreases the risk of infection in diabetic animals by modifying the interaction between intracellular lipids and phagocytic macrophages.     

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。     

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。     

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.     

云南省部分地区玉米大斑病菌生理小种鉴定

 对分离于云南省部分地区的24个大斑病单孢菌株生理小种鉴定结果表明：0号生理小种仍然是云南省的优势小种,占供试菌株的41.7％;23N号生理小种占33.3％;1号、3号及23号各占供试菌株的8.3％。     

非洲菊灰霉病抗性鉴定技术的研究

 灰霉病是非洲菊切花破坏性的病害,在采后阶段的发生和危害尤其严重,培育抗性品种具有重要意义,抗性非洲菊材料的筛选鉴定是抗性品种培育的必要技术。以生产中显著抗病和感病品种为试材,研究非洲菊灰霉病抗性鉴定技术中适宜的试验材料、灰霉菌孢子浓度、孢子侵染条件和病害评估方法。结果表明, 非洲菊花瓣正面接种病害发展快、不同抗性品种间差异明显;分生孢子的侵染萌发需要充足的氧气和营养,2μL液滴、马铃薯葡萄糖的添加使花瓣发病整齐均匀;病菌的增殖危害与接种时分生孢子和马铃薯葡萄糖的初始浓度相关,在一定浓度范围内,二者呈反比关系,其中分生孢子适宜浓度为3.0×105~1.0×106个孢子/mL,马铃薯葡萄糖的适宜浓度为3%~6%;接种花瓣放置于室温、自然光、近100%湿度条件下,接种48h是最佳的评估时间。     

不同降雨量下的毛竹林冠截留、透流及干流动态过程研究

 通过比较毛竹林降雨的林冠截留动态过程发现,当小雨量时,林冠截留曲线与林外降雨曲线吻合;当中、大雨量时,在降雨的中后期林冠截留曲线偏离林外降雨曲线,林冠截留量近乎为零。在林冠截留达到饱和的条件下,会出现截留量是负数情况;透流、干流形成和结束时间都晚于林外降雨,它们的波峰和波谷一般滞后林外降雨,透流雨相对于林外降雨振动频率低,平均振幅也较小,不同降雨量下的动态累计透流率和干流率随时间变化以多项式拟合效果较好