Hsp100 Molecular Chaperone ClpB and Its Role in Virulence of Bacterial Pathogens

This review focuses on the molecular chaperone ClpB that belongs to the Hsp100/Clp subfamily of the AAA+ ATPases and its biological function in selected bacterial pathogens, causing a variety of human infectious diseases, including zoonoses. It has been established that ClpB disaggregates and reactivates aggregated cellular proteins. It has been postulated that ClpB’s protein disaggregation activity supports the survival of pathogenic bacteria under host-induced stresses (e.g., high temperature and oxidative stress), which allows them to rapidly adapt to the human host and establish infection. Interestingly, ClpB may also perform other functions in pathogenic bacteria, which are required for their virulence. Since ClpB is not found in human cells, this chaperone emerges as an attractive target for novel antimicrobial therapies in combating bacterial infections.     

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.     

A comparison of saturated steam and superheated steam for inactivation of Escherichia coli O157:H7, Salmonella Typhimurium,and Listeria monocytogenes biofilms on polyvinyl chloride and stainless steel

This study was performed to compare the effectiveness of saturated steam (SS) and superheated steam (SHS) in the inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes biofilms on polyvinyl chloride (PVC) and stainless steel. Biofilms were formed on PVC and stainless steel coupons by using a mixture of three strains each of three foodborne pathogens at 25 °C. After biofilm development, PVC and stainless steel coupons were treated with SS at 100 °C and SHS at 125, 150, 175, and 200 °C for 5, 10, 20, and 30 s on both sides. The viable cell numbers of biofilms were significantly (p < 0.05) reduced as SHS temperature and exposure time increased. For all biofilm cells, SHS treatment resulted in an additional log reduction compared to SS treatments. After exposure to 200 °C steam for 30 s or 10 s on PVC or stainless steel, respectively, the numbers of biofilm cells were reduced to below the detection limit (1.48 log). This study demonstrated that SHS treatment effectively reduced populations of biofilm cells and reduced disinfection time compared to SS treatments and further evaluated its potential as an excellent intervention for controlling microbial biofilms and enhancing safety in the food processing industry.     

Microbial benefits and risks of raw milk cheese

Consumer preference for raw milk cheese is continually growing, owing to its more intense and varied flavor than pasteurized milk cheese. Flavor development in raw milk cheese is mainly governed by its naturally existing microbial community, which also contributes to the inhibition of food-borne pathogenic bacterial growth. Lactic acid bacteria, the dominant indigenous microorganisms of raw milk cheese, produce pathogen-inhibiting substances such as bacteriocin, organic acids, and hydrogen peroxide, and it is possible to manufacture cheese with desirable microbiological qualities. Nonetheless, outbreaks of food-borne illnesses have been linked to the consumption of raw milk cheese, and concerns have been raised regarding the microbiological safety of cheese manufactured from raw milk. Consequently, efficient and accurate methods for detecting contaminated bacterial pathogens in raw milk cheese have been promptly developed, including conventional plating, PCR-based technology, and immunoassay-integrated methods. The microbiological risk of the cheese can be reduced by proper ripening processing. However, additionally, hygiene in the environments for milk production and cheesemaking and the post-manufacturing stage needs to be constantly microbiologically monitored.     

2016—2020年郴州市毒蕈中毒事件流行病学特征分析

目的 通过对2016—2020年郴州市毒蕈中毒事件的流行病学特征进行描述性分析,为制定毒蕈中毒预防控制措施提供科学依据。方法 收集和整理2016—2020年郴州市通过“食源性疾病事件报告系统”上报的毒蕈中毒事件资料,对事件发生的时间、地区、人群特征以及毒蕈来源、种类进行描述性分析。结果 2016—2020年郴州市共报告毒蕈中毒事件131起,发病461例,死亡6例,病死率为1.3%;事件起数最多的是2020年,中毒发生时间集中在5～10月;事件报告起数和发病人数居前3位的依次是嘉禾县、北湖区和宜章县,多发生在农村地区;家庭是毒蕈中毒高发场所,毒蕈主要来源于市民自行采摘,也有部分从市场购买;已知毒蕈种类的中毒事件中的事件起数和发病例数前3位的依次为日本红菇、大青褶伞、近江粉褶菌,死亡病例由灰花纹鹅膏和亚稀褶红菇中毒引起。结论 应针对特定时间、地点和人群进行毒蕈中毒宣传教育,同时应加强对野生蕈市场销售的监管,开展毒蕈中毒的相关研究,以科学预防毒蕈中毒的发生。     

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.     

The Periplasmic Oxidoreductase DsbA Is Required for Virulence of the Phytopathogen Dickeya solani

In bacteria, the DsbA oxidoreductase is a crucial factor responsible for the introduction of disulfide bonds to extracytoplasmic proteins, which include important virulence factors. A lack of proper disulfide bonds frequently leads to instability and/or loss of protein function; therefore, improper disulfide bonding may lead to avirulent phenotypes. The importance of the DsbA function in phytopathogens has not been extensively studied yet. Dickeya solani is a bacterium from the Soft Rot Pectobacteriaceae family which is responsible for very high economic losses mainly in potato. In this work, we constructed a D. solani dsbA mutant and demonstrated that a lack of DsbA caused a loss of virulence. The mutant bacteria showed lower activities of secreted virulence determinants and were unable to develop disease symptoms in a potato plant. The SWATH-MS-based proteomic analysis revealed that the dsbA mutation led to multifaceted effects in the D. solani cells, including not only lower levels of secreted virulence factors, but also the induction of stress responses. Finally, the outer membrane barrier seemed to be disturbed by the mutation. Our results clearly demonstrate that the function played by the DsbA oxidoreductase is crucial for D. solani virulence, and a lack of DsbA significantly disturbs cellular physiology.     

复合侵染的蚕豆黄花叶病病原诊断

应用透射电镜、ELISA和RT-PCR技术对浙江丽水的蚕豆黄化花叶病株进行了病原诊断,电镜负染色发现病株汁液中存在线状和球状两种病毒粒子,线状粒子长度在600nm～800nm的占73%,球状粒子直径约26nm;超薄切片观察到细胞质中存在Ⅱ型风轮状内含体及大量电子致密无定型体,还有线粒体增生聚集和膜结构的增生,两类细胞病变现象出现在相邻细胞中;用BBWV 2的单克隆抗体对病汁液ELISA检测结果为阳性反应;用Potyvirus通用引物进行RT-PCR检测有1.7kb特异性扩增片段产生.根据以上结果诊断病原为蚕豆萎焉病毒2（BBWV 2）和菜豆黄花叶病毒（BYMV）,二者为复合侵染.     

Benchmarking Long-Read Assemblers for Genomic Analyses of Bacterial Pathogens Using Oxford Nanopore Sequencing

Oxford Nanopore sequencing can be used to achieve complete bacterial genomes. However, the error rates of Oxford Nanopore long reads are greater compared to Illumina short reads. Long-read assemblers using a variety of assembly algorithms have been developed to overcome this deficiency, which have not been benchmarked for genomic analyses of bacterial pathogens using Oxford Nanopore long reads. In this study, long-read assemblers, namely Canu, Flye, Miniasm/Racon, Raven, Redbean, and Shasta, were thus benchmarked using Oxford Nanopore long reads of bacterial pathogens. Ten species were tested for mediocre- and low-quality simulated reads, and 10 species were tested for real reads. Raven was the most robust assembler, obtaining complete and accurate genomes. All Miniasm/Racon and Raven assemblies of mediocre-quality reads provided accurate antimicrobial resistance (AMR) profiles, while the Raven assembly of Klebsiella variicola with low-quality reads was the only assembly with an accurate AMR profile among all assemblers and species. All assemblers functioned well for predicting virulence genes using mediocre-quality and real reads, whereas only the Raven assemblies of low-quality reads had accurate numbers of virulence genes. Regarding multilocus sequence typing (MLST), Miniasm/Racon was the most effective assembler for mediocre-quality reads, while only the Raven assemblies of Escherichia coli O157:H7 and K. variicola with low-quality reads showed positive MLST results. Miniasm/Racon and Raven were the best performers for MLST using real reads. The Miniasm/Racon and Raven assemblies showed accurate phylogenetic inference. For the pan-genome analyses, Raven was the strongest assembler for simulated reads, whereas Miniasm/Racon and Raven performed the best for real reads. Overall, the most robust and accurate assembler was Raven, closely followed by Miniasm/Racon.     

食源性微生物MRSA及其检测方法在食品安全中的研究进展

食品安全在世界卫生组织的定义是指所有食品中有毒、有害物质对人体健康影响的公共卫生问题。2008年"三聚氰胺"事件后,在近几年的全国两会上,食品安全问题连续成为焦点话题;而同期全国人大常委会通过的《食品安全法》,更显示了食品安全的严重与令人担忧,其监测与控制显得迫在眉睫。作为典型的食源性微生物,金葡菌尤其是耐甲氧西林金黄色葡萄球菌(Methicillin-resistant Staphylococcus aureus,MRSA)其新型的耐药特点显示了细菌耐药性的发展和进化可能是由于大量抗生素在畜牧业中的滥用而造成的。由于抗菌药物的研制周期跟不上耐药性出现的速度,使得细菌治疗面临越来越严峻的考验,甚至出现无药可用的情况,严重威胁人类健康。本文结合食品安全抗生素滥用问题与最近出现的超级细菌耐药问题,根据在相关领域多年的研究数据,对常见的食源性微生物MRSA及其检测方法在食品安全中的研究进展进行综述。