Microbial enrichment and multiplexed microfiltration for accelerated detection of Salmonella in spinach

To attain Salmonella detection thresholds in spinach suspensions using enrichment media requires at least 24 hr. Separation and concentration of selected microorganisms via microfiltration and microfugation reduce time for sample preparation, especially when working with large volumes of vegetable suspensions. This facilitates accelerated detection of Salmonella in spinach suspensions, and may contribute to effectively monitoring this pathogen before it reaches the consumer. We report a microfiltration-based protocol for accelerated sample preparation to concentrate and recover ≤1 colony forming unit (CFU) Salmonella/g pathogen-free spinach. Store-bought samples of spinach and a spinach plant subjected to two environmental conditions (temperature and light exposure) during its production were tested. The overall procedure involves extraction with buffer, a short enrichment step, prefiltration using a nylon filter, crossflow hollow fiber microfiltration, and retentate centrifugation to bring microbial cells to detection levels. Based on 1 CFU Salmonella/g frozen spinach, and a Poisson distribution statistical analyses with 99% probability, we calculated that 3 hr of incubation, when followed by microfiltration, is sufficient to reach the 2 log concentration required for Salmonella detection within 7 hr. Longer enrichment times (5 hr or more) is needed for concentrations lower than 1 CFU Salmonella/g of ready to eat spinach. The recovered microbial cells were identified and confirmed as Salmonella using both polymerase chain reaction (PCR) and plating methods. Different environmental conditions tested during production did not affect Salmonella viability; this demonstrated the broad adaptability of Salmonella and emphasized the need for methods that enable efficient monitoring of production for the presence of this pathogen.     

Global distribution of plasmid-mediated colistin resistance mcr gene in Salmonella: A systematic review

This systematic review focuses on obtaining the most relevant information from multiple studies that detected a mobilized colistin resistance mcr gene in Salmonella for a better comprehension of its global distribution. A group of strategic and systematic keywords were combined to retrieve research data on the detection frequency of the mcr gene globally from four database platforms (Google Scholar, Science Direct, PubMed and Scielo). Forty-eight studies attended all the eligibility criteria and were selected. China was the country with the highest frequency of Salmonella strains with the mcr gene, and Europe exhibited a wide diversity of countries with positive mcr strains. In addition, animals and humans carried the highest frequency of positive strains for the mcr gene. Salmonella Typhimurium was the most frequent serovar carrying the mcr gene. Apparently, colistin overuse in animal husbandry has increased the selective pressure of antimicrobial resistance, resulting in the emergence of a plasmid-mediated colistin resistance mcr gene in China. The mcr-positive Salmonella strains are recently predominant worldwide, which is probably due to the capacity of this gene to be swiftly horizontally transmissible. The transmission ability of mcr-positive Salmonella strains to humans through the consumption of contaminated animal-based food is a public health concern.     

伤寒沙门菌非编码RNA617的分子鉴定及其对生物膜形成的影响研究

本研究旨在探讨伤寒沙门菌(Salmonella enterica serovar Typhi, S. Typhi)中非编码RNA617(non-coding RNA617,ncRNA617)的分子特性,并研究其对生物膜形成的影响及作用机制。采用Northern blot方法检测ncRNA617的表达,通过cDNA 5’末端快速扩增技术(5’-rapid amplification of cDNA end,5’RACE)和逆转录-聚合酶链式反应(reverse transcriotion-polymerase chain reaction,3’RT-PCR)实验分析ncRNA617可能的转录起始位点和终止位点;构建ncRNA617缺陷菌株、回补菌株和过表达菌株等相关菌株,通过生物膜形成实验,观察ncRNA617对伤寒沙门菌生物膜形成的影响,并用实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qPCR)分析生物膜形成相关基因表达水平的变化,综合运用生物信息学方法预测ncRNA617和差异基因的结合区域,初步分析ncRNA617发挥调控作用的机制。结果显示,伤寒沙门菌确有ncRNA617的表达,长度约300 nt,其转录起始位点位于mig-14终止密码子下游967 nt处,终止位点位于t2681起始密码子上游 2 378～2 560 nt处。与野生对照菌株相比,ncRNA617缺陷菌株生物膜形成能力增强(P<0.05),回补菌株的生物膜形成能力恢复至野生菌株水平,过表达菌株的生物膜形成能力有所下降(P<0.05)。qPCR结果表明,ncRNA617可负向调控多个生物膜形成相关基因的转录表达水平(P<0.05)。经生物信息学方法预测发现,ncRNA617与差异基因有不同的结合区域。本研究结果提示,ncRNA617在伤寒沙门菌中存在,其长度约270～452 nt。ncRNA617可能通过靶向结合生物膜形成相关基因下调基因表达,从而负向调控伤寒沙门菌生物膜的生成。