副溶血性弧菌多克隆抗体制备及应用

以热灭活副溶血性弧菌(Vibrio parahaemolyticus,VP ATCC17802)免疫新西兰大白兔制备多克隆抗体,用间接酶联免疫吸附法测定抗血清效价,纯化后的多克隆抗体效价为512000。该抗体与5株副溶血性弧菌均能特异性结合,但与其他12株非副溶血性弧菌无交叉反应。在此基础之上建立了间接竞争酶联免疫吸附法(indirect competitive enzyme-linked immunosorbent assay,ic-ELISA),并优化了抗原抗体浓度、二抗浓度、封闭液等条件。结果表明,该法的线性范围为5×10^4~107CFU/m L,线性回归方程为y=0.21x-0.74(R2=0.99),IC50为106CFU/m L,最低检出限为1.7×10^4CFU/m L,板内变异系数为2.61%~4.15%,板间变异系数为4.71%~8.74%。用建立的ic-ELISA检测市场中鱼、贝类养殖水和实验室用水中的VP,实验结果与国标检测方法的结果一致。该方法快速便捷,灵敏度高,准确性好,为进一步研究VP的快速检测奠定了基础。     

Recent developments in nanotechnology transforming the agricultural sector: a transition replete with opportunities

The applications and benefits of nanotechnology in the agricultural sector have attracted considerable attention, particularly in the invention of unique nanopesticides and nanofertilisers. The contemporary developments in nanotechnology are acknowledged and the most significant opportunities awaiting the agriculture sector from the recent scientific and technical literature are addressed. This review discusses the significance of recent trends in nanomaterial‐based sensors available for the sustainable management of agricultural soil, as well as the role of nanotechnology in detection and protection against plant pathogens, and for food quality and safety. Novel nanosensors have been reported for primary applications in improving crop practices, food quality, and packaging methods, thus will change the agricultural sector for potentially better and healthier food products. Nanotechnology is well‐known to play a significant role in the effective management of phytopathogens, nutrient utilisation, controlled release of pesticides, and fertilisers. Research and scientific gaps to be overcome and fundamental questions have been addressed to fuel active development and application of nanotechnology. Together, nanoscience, nanoengineering, and nanotechnology offer a plethora of opportunities, proving a viable alternative in the agriculture and food processing sector, by providing a novel and advanced solutions. © 2017 Society of Chemical Industry     

Enhanced phagocytosis of Aggregatibacter actinomycetemcomitans cells by macrophages activated by a probiotic Lactobacillus strain

The activation of phagocytosis is one important approach to clearing pathogenic cells in a host. This study evaluated the ability of probiotic lactobacilli to induce phagocytic activity as well as the clearance of a periodontal pathogen, Aggregatibacter actinomycetemcomitans. First, the activation of phagocytosis was found by using lyophilized dead cells. Probiotic Lactobacillus strains significantly enhanced the phagocytic activity of macrophage cells, indicating that the probiotic lactobacilli have a remarkable ability to stimulate the macrophages. Essentially, 3 Lactobacillus strains tested did not have any critical toxic effect on the murine macrophage, and Lactobacillus johnsonii NBRC 13952 showed the least cytotoxic effect on the RAW264.7 macrophages. The expression of classically activated macrophage markers, IL-1β, and cluster of differentiation 80 increased by L. johnsonii NBRC 13952; however, there was no significant difference for IL-18. The highest phagocytic activity by macrophages was found in a condition in which the macrophage activated by L. johnsonii NBRC 13952 functions to kill the cells of A. actinomycetemcomitans. Correlating with the result, a high amount of hypodiploid DNA (SubG1) was detected from the macrophage cells stimulated by L. johnsonii NBRC 13952. Taken together, the results suggest that macrophages activated by the Lactobacillus strain can facilitate the phagocytosis of A. actinomycetemcomitans cells by linking with enhanced apoptotic activities. In conclusion, L. johnsonii NBRC 13952 has a certain role in activating the RAW264.7 macrophages, thereby counteracting the infection of A. actinomycetemcomitans.     

Efficacy of thyme oil-alginate-based coating in reducing foodborne pathogens on fresh-cut apples

In this study, the inhibition of an alginate-based edible coating (EC) containing thyme oil (0.05%, 0.35% and 0.65%) was evaluated against Listeria monocytogenes, Salmonella Typhimurium, Staphylococcus aureus and Escherichia coli O157:H7 inoculated onto fresh-cut apples. To investigate the antibacterial mechanism of thyme oil, the constituent compounds of that were analysed by gas chromatography-mass spectrometry (GC-MS), and the cellular damage of pathogens was observed by scanning electron microscopy (SEM). Results showed that alginate-based EC containing thyme oil effectively inhibited the growth of pathogens on fresh-cut apples. GC-MS analysis revealed thymol (47.23%) as the major compounds in thyme oil. SEM showed that the cell membrane of foodborne pathogens was damaged by thyme oil, causing their inactivation. Treatment with alginate-based EC containing 0.05% thyme oil preserved the sensory characteristics of fresh-cut apples. Therefore, using alginate-based EC with thyme oil may represent a potential approach to preserve and enhance the safety of fresh-cut apples.     

Antimicrobial activity of acid-hydrolyzed Citrus unshiu peel extract in milk

Citrus fruit (Citrus unshiu) peels were extracted with hot water and then acid-hydrolyzed using hydrochloric acid. Antimicrobial activities of acid-hydrolyzed Citrus unshiu peel extract were evaluated against pathogenic bacteria, including Bacillus cereus, Staphylococcus aureus, and Listeria monocytogenes. Antilisterial effect was also determined by adding extracts at 1, 2, and 4% to whole, low-fat, and skim milk. The cell numbers of B. cereus, Staph. aureus, and L. monocytogenes cultures treated with acid-hydrolyzed extract for 12 h at 35°C were reduced from about 8 log cfu/mL to <1 log cfu/mL. Bacillus cereus was more sensitive to acid-hydrolyzed Citrus unshiu peel extract than were the other bacteria. The addition of 4% acid-hydrolyzed Citrus unshiu extracts to all types of milk inhibited the growth of L. monocytogenes within 1 d of storage at 4°C. The results indicated that Citrus unshiu peel extracts, after acid hydrolysis, effectively inhibited the growth of pathogenic bacteria. These findings indicate that acid hydrolysis of Citrus unshiu peel facilitates its use as a natural antimicrobial agent for food products.     

常见食源性致病菌胞外代谢轮廓分析

寻找常见食源性致病菌间特征性代谢产物,是研发食品安全快速高效监控技术的基础。本文直接利用常见食源性致病菌发酵液冻干,经硅烷化试剂衍生,并采用气相色谱-质谱技术(GC-MS)对代谢产物进行分析,同时进行NIST11谱图库检索并分类,并对数据进行热图和主成分(PCA)分析。研究发现,发酵液中有大量有机酸、醇类和胺类等物质产生,且菌种间各代谢产物种类和相对含量差异显著;同时各菌间含有丰富的特有代谢产物,其中部分特有代谢产物在其他菌株中未见报道。通过热图和PCA分析各菌株胞外代谢轮廓可知,24 h时各菌株间能够明显区分,同时去除糖类和氨基酸后,PCA区分鉴定效果明显提高,尤其在24 h时最好。研究表明,胞外代谢轮廓分析可以用于寻找常见食源性致病菌生物标志物和菌种区分鉴定,同时部分菌种间特有的代谢产物有望成为常见食源性致病菌潜在生物标志物。     

DNA微阵列在食品营养与安全中的应用与展望

DNA微阵列是一种高通量基因检测和分析技术,主要概括介绍DNA微阵列在营养基因组学、食源性病原体检测、转基因食品和饮食补充剂安全性评价中的应用。     

2013年山东省食源性疾病暴发事件 流行病学特征分析

目的分析2013年山东省食源性疾病暴发事件的流行病学特征,为制定食源性疾病预防控制措施提供依据。方法对2013年通过国家食源性疾病暴发报告系统上报的30起食源性疾病暴发事件进行整理分析。结果 2013年共发生食源性疾病暴发事件30起,发病人数654人,死亡2人。食源性疾病暴发事件的主要发病季节是第三季度,全年食源性疾病暴发主要发生在4~10月份,其中以8月份最高;16~60岁人群是暴发事件发生的主要人群;饮食服务单位是发生食源性疾病暴发事件的主要场所,其次是集体食堂;加工不当与交叉污染是引起食源性疾病暴发事件的主要原因:微生物是引起食源性疾病暴发事件发生的主要致病因素,不同致病因素之间导致的罹患率不同(P0.05);引起食源性疾病暴发事件的主要原因食品是混合食品。结论加强对高发季节、高发因素、高发污染环节的监控;加强食源性疾病暴发事件的调查处置;加大防控食源性疾病暴发事件的宣传力度等,是预防和控制食源性疾病暴发事件的有效措施。     

多重PCR法检测鲜切哈密瓜中3种食源性致病菌

建立可同时检测鲜切哈密瓜中的单增李斯特菌、鼠伤寒沙门氏菌和大肠杆菌O157:H7的多重聚合酶链式反应(polymerase chain reaction,PCR)检测方法。根据单增李斯特菌inl A基因、鼠伤寒沙门氏菌inv A基因、大肠杆菌O157:H7的wzy基因设计3对特异性引物。对多重PCR反应体系中的引物浓度、退火温度、Mg2+浓度、d NTP浓度进行了优化,并确定适宜的多重PCR反应体系及反应条件。结果表明:25μL反应体系,10×PCR buffer为2.5μL,Mg Cl2(25 mmol/L)为3.5μL,d NTPs(2.5 mmol/L)为2μL,inl A基因上下游引物(5μmol/L)为1μL,inv A基因上下游引物(5μmol/L)为1μL,wzy基因上下游引物(5μmol/L)为2μL,单增李斯特菌DNA模板为1μL,鼠伤寒沙门氏菌DNA模板为1μL,大肠杆菌O157:H7 DNA模板为1μL,ex Taq DNA聚合酶(5 U/L)为0.3μL,加dd H2O补足25μL。反应条件为95℃预变性3 min;94℃变性30 s,53.9℃退火30 s,72℃延伸30 s,32个循环;72℃延伸10 min。鲜切哈密瓜中人工接种的目标菌的灵敏度为单增李斯特菌2.7×104 CFU/g,大肠杆菌O157:H7 3.3×104 CFU/g以及鼠伤寒沙门氏菌3.8×104 CFU/g。该技术可为快速检测鲜切哈密瓜中病原菌污染度及其控制提供参考依据。     

射频杀菌条件下黑胡椒颗粒中沙门氏菌的替代菌研究

为探究热杀菌过程中屎肠球菌NRRL B-2354作为沙门氏菌ATCC 14028替代菌的可行性,通过等温实验测定黑胡椒颗粒中屎肠球菌NRRL B-2354和沙门氏菌ATCC 14028的耐热性参数,并对黑胡椒颗粒（A_w=0.65）开展射频杀菌实验验证。等温实验结果表明,黑胡椒颗粒中屎肠球菌的耐热性高于沙门氏菌。射频杀菌实验中,将2 g接种微生物的黑胡椒颗粒真空包装后置于45 g黑胡椒中心位置,在40.68 MHz射频场中加热15 min,黑胡椒颗粒中沙门氏菌的减少量相较于屎肠球菌更多,证明在射频加热中屎肠球菌的耐热性高于沙门氏菌,可作为黑胡椒颗粒杀菌过程中沙门氏菌的合适替代菌。