Leptosphaeria maculans-Brassica napus Battle: A Comparison of Incompatible vs. Compatible Interactions Using Dual RNASeq

Leptosphaeria maculans causes blackleg disease, which is one of the most destructive diseases of canola (Brassica napus L.). Due to the erosion of the current resistance in B. napus, it is pivotal to introduce new resistant genotypes to the growers. This study evaluated the potential of Rlm7 gene as resistance to its corresponding avirulence AvrLm7 gene is abundant. The Rlm7 line was inoculated with L. maculans isolate with AvrLm7; UMAvr7; and the CRISPR/Cas9 knockout AvrLm7 mutant, umavr7, of the same isolate to cause incompatible and compatible interactions, respectively. Dual RNA-seq showed differential gene expressions in both interactions. High expressions of virulence-related pathogen genes-CAZymes, merops, and effector proteins after 7-dpi in compatible interactions but not in incompatible interaction—confirmed that the pathogen was actively virulent only in compatible interactions. Salicyclic and jasmonic acid biosynthesis and signaling-related genes, defense-related PR1 gene (GSBRNA2T00150001001), and GSBRNA2T00068522001 in the NLR gene family were upregulated starting as early as 1- and 3-dpi in the incompatible interaction and the high upregulation of those genes after 7-dpi in compatible interactions confirmed the early recognition of the pathogen by the host and control it by early activation of host defense mechanisms in the incompatible interaction.     

Short communication: Microbiological quality of raw cow milk and its association with herd management practices in Northern China

Contamination of raw milk with bacterial pathogens is potentially hazardous to human health. The aim of this study was to evaluate the total bacteria count (TBC) and presence of pathogens in raw milk in Northern China along with the associated herd management practices. A total of 160 raw milk samples were collected from 80 dairy herds in Northern China. All raw milk samples were analyzed for TBC and pathogens by culturing. The results showed that the number of raw milk samples with TBC <2 × 10⁶ cfu/mL and <1 × 10⁵ cfu/mL was 146 (91.25%) and 70 (43.75%), respectively. A total of 84 (52.50%) raw milk samples were Staphylococcus aureus positive, 72 (45.00%) were Escherichia coli positive, 2 (1.25%) were Salmonella positive, 2 (1.25%) were Listeria monocytogenes positive, and 3 (1.88%) were Campylobacter positive. The prevalence of S. aureus was influenced by season, herd size, milking frequency, disinfection frequency, and use of a Dairy Herd Improvement program. The TBC was influenced by season and milk frequency. The correlation between TBC and prevalence of S. aureus or E. coli is significant. The effect size statistical analysis showed that season and herd (but not Dairy Herd Improvement, herd size, milking frequency, disinfection frequency, and area) were the most important factors affecting TBC in raw milk. In conclusion, the presence of bacteria in raw milk was associated with season and herd management practices, and further comprehensive study will be powerful for effectively characterizing various factors affecting milk microbial quality in bulk tanks in China.     

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。     

活的非可培养状态副溶血性弧菌实时荧光逆转录PCR检测方法的建立及其毒力研究

目的 建立检测活的非可培养(VBNC)状态副溶血性弧菌的荧光逆转录PCR方法并研究其毒力基因表达.方法 将副溶血性弧菌AS079菌株添加到陈化海水中,4℃冰箱内培养,使其进入VBNC状态,针对其管家基因、鉴定基因和毒力基因分别设计实时荧光逆转录PCR引物,通过不同的PCR反应程序和引物浓度组合试验,摸索最佳反应体系条件,用于VBNC状态副溶血性弧菌的检测和毒力研究.结果 用所建立的检测VBNC状态副溶血性弧菌的实时荧光逆转录PCR方法,对接种于陈化海水培养的不同时期的AS079副溶血性弧菌进行荧光定量逆转录PCR扩增,结果显示,随着培养时间的延长,毒力基因tdh2和鉴定基因toxR表达水平持续下降,但即使细菌进入VBNC状态,这两个基因也能够得到很好的扩增,说明以toxR基因和tdh2基因对进入VBNC状态的副溶血性弧菌进行检测和毒力研究方法可行.灵敏度试验表明,鉴定基因toxR的最低检测限可达到48 cfu/ml,研究毒力基因tdh2的表达需要细菌浓度至少为4.8×102cfu/ml,同时试验证明此方法与其他相近食源性致病菌无交叉反应.结论 该实时荧光逆转录PCR方法检测快速、特异性强、敏感度高,适用于VBNC状态副溶血性弧菌的检测和毒力分析.     

两性霉素B在治疗眼部真菌感染的应用

目的概述两性霉素B治疗眼科真菌感染的作用和价值。方法参考近年来的文献,论述两性霉素B的作用机制及在眼科中应用。结果两性霉素B是抗菌谱广、抗菌作用强的抗真菌药物。结论两性霉素B是临床上不可缺少的抗真菌药物,两性霉素B脂质体在治疗眼部真菌感染具有良好的应用前景。     

Enhanced antimicrobial activity of nisin-loaded liposomal nanoparticles against foodborne pathogens

This study was designed to evaluate the prolonged antimicrobial stability of nisin-loaded liposome (LipoN) nanoparticles against Listeria monocytogenes and Staphylococcus aureus. The sizes of bare liposomes and LipoN were uniformly distributed between 114 and 125 nm. The nanoparticles were homogeneously dispersed in water with less than 0.2 of polydispersity index. The zeta potential value of LipoN was +17.1 mV due to the positive charged nisin, attaining 70% of loading efficiency. The minimum inhibitory concentration of LipoN against L. monocytogenes and S. aureus was 320 international unit/mL. The LipoN significantly enhanced the antimicrobial stability in brain heart infusion agar compared to free nisin. The numbers of L. monocytogenes and S. aureus exposed to LipoN were effectively reduced by more than 6 log colony-forming unit/mL after 48 and 72 h of incubation, respectively. These results provide useful information for the development of antimicrobial delivery system to improve food safety.     

基因芯片法与常规检测法在食源性疾病中的应用对比分析

目的对比分析基因芯片法与常规检测法在食源性疾病中的应用效果。方法收集2016年4月到2018年8月收治的诊断为食源性疾病的患者98例为研究对象,收集患者的新鲜粪便,根据检测方法不同分为对照组与观察组,观察组应用基因芯片法进行病原菌的检测,对照组则应用常规培养法进行病原菌的检测。,观察比较两组的病原菌检测阳性率和检测时间等相关指标。结果在98例患者中,观察组采用基因芯片法检测出病原菌58例(副溶血弧菌9例,痢疾杆菌32例,沙门菌10例,大肠埃希菌2例,金黄色葡萄球菌5例),病原菌检出率为58.2%。对照组采用常规培养法检测出病原菌32例(副溶血弧菌5例,痢疾杆菌19例,沙门菌4例,金黄色葡萄球菌4例),病原菌检出率为32.7%。基因芯片法和培养法对病原菌、痢疾杆菌的检出率差异存在统计学意义(P0.05)。观察组采用基因芯片法用于食源性疾病患者的病原菌检测时间明显短于常规培养法,差异有统计学意义(P0.05)。结论基因芯片技术能够快速检出食源性疾病病原菌,可以替代常规培养法对病原菌进行检测,在食源性疾病的早期诊断和治疗中具有重要的临床价值。     

Categorizing food-related illness: Have we got it right?

Since the 1950s food safety hazards have been categorized simply as (micro) biological, chemical or physical hazards with no clear differentiation between those that cause acute and chronic harm. Indeed international risk assessment methods, including hazard analysis critical control point (HACCP) use these criteria. However, the spectrum of food related illness continues to grow now encompassing food allergy and intolerance, obesity, type 2 diabetes, stroke, heart disease, cancer as well as food poisoning, foodborne illness and food contamination. Therefore over a half-century later is this the time to redefine the spectrum of what constitutes food related illness? This paper considers whether such “redefinition” of food related intoxicating and infectious agents would provide more targeted policy instruments and lead to better risk assessment and thus mitigation of such risk within the food supply chain.     

The mitochondrial genome from the thermal dimorphic fungus Paracoccidioides brasiliensis

We present here the sequence of the mitochondrial DNA of the pathogenic thermodimorphic fungus Paracoccidioides brasiliensis, agent of an endemic disease in most South American countries. The sequenced genome has 71 334 bp and is organized as a circular molecule with two gaps of unknown size flanking the middle exon of the nad5 gene. We located genes coding for the three subunits of the ATP synthase (atp6, atp8 and atp9), the apocytochrome b (cob), three subunits of the cytochrome c oxidase enzyme complex (cox1, cox2 and cox3), seven subunits of the reduced nicotinamide adenine dinucleotide ubiquinone oxidoreductase (nad1, nad2, nad3, nad4, nad5, nad6 and nad4L) and the large (rnl) and small (rns) subunits of ribosomal RNA. Two maturases and a ribosomal protein (rms5) are located inside introns. Twenty-five tRNAs were identified with acceptors for all 20 amino acids. Seven polypurine/polypyrimidine tracts (140-240 bp) have been found in this genome. All genes are in the same orientation over the genome, while their order is closest to the mitochondrial genomes from Penicillium marneffei and Aspergillus nidulans.     

广州市白云口岸航空食品中金黄色葡萄球菌凝固酶基因分型及肠毒素基因研究

目的对2013—2015年从广州市白云口岸航空食品中分离的金黄色葡萄球菌进行基因分型研究,为食源性金黄色葡萄球菌分子溯源提供基础数据。方法以血浆凝固酶和肠毒素为目标基因,采用聚合酶链式反应(PCR)方法对9株金黄色葡萄球菌进行基因分型,其中6株为航空食品分离株,1株为配餐车间大门手拭分离株,2株为标准菌株。肠毒素基因检测包括5种传统肠毒素基因(sea、seb、sec、sed、see)和6种新型肠毒素基因(ser、seg、seh、sei、sej、sep)。结果 6株航空食品分离株的血浆凝固酶基因扩增分型结果为2个PCR型,酶切后得3种亚型;肠毒素基因检测结果显示有2株航空食品分离株含有肠毒素基因,检出率为33.3%(2/6),检出的基因为2种传统肠毒素基因(sec、sed)和4种新型肠毒素基因(ser、seg、sei、sej),均同时携带2种以上肠毒素基因。结论血浆凝固酶基因扩增分型结果显示,不同时间、不同采集地点存在相同的基因型,提示金黄色葡萄球菌存在交叉污染的可能性;航空食品分离株共检出6种肠毒素基因,提示金黄色葡萄球菌基因型多样性,应加强其他新型肠毒素基因检测。