Antimicrobial resistance and co-selection phenomenon in Listeria spp. recovered from food and food production environments

Antimicrobial resistance (AMR), co-selection phenomenon, and the relationship between reduced susceptibility (RSC) to ciprofloxacin (CIP) and resistance to other antimicrobials in Listeria spp. (n = 103) recovered from food processing environments (FPE) and food were investigated. Resistance of Listeria monocytogenes and other listeriae, respectively, to cefoxitin (FOX; 98% vs. 88%), CIP (7% vs. 4%), clindamycin (CLI; 33% vs. 59%) and tetracycline (6% vs. 8%) was observed, as was RSC to CIP (67% vs. 57%) and CLI (65% vs. 41%). L. monocytogenes also possessed RSC to linezolid (LZD; 6%), rifampicin (2%) and streptomycin (6%), with other listeriae displaying RSC to chloramphenicol (4%). L. monocytogenes serotype 1/2a (90%) isolates were more frequently resistant or possessed RSC to CIP compared to serotype 4b (55%) (p = 0.015). When eight strains were experimentally adapted to high concentrations of CIP, co-selection occurred as MICs to benzalkonium chloride (BAC) increased (n = 5), gentamicin MICs remained the same (n = 6) or increased 2-fold (n = 2), and led to RSC to LZD (n = 1) and resistance to CLI (n = 8). Overall, levels of resistance/RSC to CIP in food chain isolates, particularly 1/2a, are concerning. Further, reduced sensitivity to disparate antimicrobials following CIP exposure highlights the need for increased knowledge of co-selection phenomenon linked with antimicrobial agents.     

Spoilage of refrigerated (4 °C) Litopenaeus vannamei: cooperation between Shewanella species and contribution of cyclo‐(L‐Pro‐L‐Leu)‐dependent quorum sensing

The spoilage potential of Shewanella putrefaciens and S. baltica isolated from spoiled refrigerated Litopenaeus vannamei was evaluated by in vitro assays for trimethylamine oxide reduction, extracellular hydrolytic enzymes and biofilm formation, and in vivo inoculation into surface‐sterilised shrimp followed by microbial, biochemical and sensory analyses during storage for 5 days at 4 °C. S. baltica displayed higher spoilage potential than S. putrefaciens both in vitro and in vivo. Shrimp co‐inoculated with them had one‐day shorter shelf life than those mono‐inoculated, based on the results of bacterial density, volatile base nitrogen, trimethylamine, volatile organic compounds and sensory analysis, which strongly suggests cooperation of Shewanella species in shrimp spoilage. Exogenous cyclo‐(L‐Pro‐L‐Leu) boosted bacterial growth, extracellular protease and collagenase activities, and biofilm formation of S. putrefaciens and S. baltica at least before they entered the stationary phase, indicating that cyclo‐(L‐Pro‐L‐Leu)‐dependent quorum sensing, a recently suggested communication mechanism between them, contributes to the cooperation.     

Antimicrobial packaging of raw beef,pork and turkey using silver‐zeolite incorporated into the material

This study determined the efficacy of silver‐zeolite impregnated into wrapping paper to reduce the bacterial growth on raw beef, pork and turkey cuts. This was compared with that of regular butcher paper. The samples were inoculated with Pseudomonas putida (psychrotrophic spoilage bacterium) and stored on 4% silver‐zeolite and regular butcher paper for up to 4 days at 4 °C and 10 °C. Results showed that P. putida on the beef, pork and turkey samples did not increase in numbers after exposure to all paper packaging at 4 °C during the 4 days of storage. At 10 °C storage temperature, logarithmic growth patterns for the organisms were seen on all paper packaging. However, the growth rate was slower for the organisms on the silver‐zeolite paper. Storage on the silver‐zeolite paper accounted for one log cfu/sample mean reduction in viable cell count for the beef, pork and turkey samples when compared with the samples stored on the butcher paper at 3 days.     

Chemical Signals of Synthetic Disaccharide Derivatives Dominate Rhamnolipids at Controlling Multiple Bacterial Activities

Microbes secrete molecules that modify their environment. Here, we demonstrate a class of synthetic disaccharide derivatives (DSDs) that mimics and dominates the activity of naturally secreted rhamnolipids by Pseudomonas aeruginosa. The DSDs exhibit the dual function of activating and inhibiting the swarming motility through a concentration‐dependent activity reversal that is characteristic of signaling molecules. Whereas DSDs tethered with a saturated farnesyl group exhibit inhibition of both biofilm formation and swarming motility, with higher activities than rhamnolipids, a saturated farnesyl tethered with a sulfonate group only inhibits swarming motility but promote biofilm formation. These results identified important structural elements for controlling swarming motility, biofilm formation, and bacterial adhesion and suggest an effective chemical approach to control intertwined signaling processes that are important for biofilm formation and motilities.     

海洋环境中假单胞菌对聚氨酯清漆涂层分解的影响

目的 研究在海洋环境中假单胞菌对聚氨酯清漆涂层分解作用的影响。方法 将聚氨酯清漆涂层样品分别浸泡于无菌海水与含假单胞菌的海水中,对浸泡环境不同的样品利用电化学阻抗谱（EIS）评价涂层防腐性能,使用扫描电子显微镜（SEM）和傅里叶变换红外光谱（FTIR）,观察对比涂层的表面形貌,并表征其分子结构变化。结果 1 h~35 d时,在无菌海水及假单胞菌海水中,样品Nyquist图的容抗弧直径以及Bode图的低频端阻抗模量均减小。与此同时,假单胞菌海水中浸泡样品的Nyquist图容抗弧直径与Bode图低频端阻抗模量的减小量明显大于无菌海水中的减小量。无菌海水浸泡的等效电路在1~48 h时为1个时间常数,5~35 d时增加为2个时间常数;假单胞菌海水浸泡的等效电路在1~48 h时只有1个时间常数,5~29 d时为2个时间常数,35 d时增加为3个时间常数。浸泡36 h时,无菌海水中浸泡样品的涂层电阻值为8.23× 107 Ω?cm2,而假单胞菌海水中样品的涂层电阻为5.14×107 Ω?cm2;至35 d时,无菌海水中样品的涂层电阻降为5.61×106 Ω?cm2,假单胞菌海水中样品的涂层电阻降至7.03×105 Ω?cm2。假单胞菌海水中样品的涂层电阻在36 h~35 d的减小量明显大于无菌海水中的减小量。由SEM结果可以观察到,浸泡30 d后,无菌海水中浸泡样品的表面光滑完整,而在含假单胞菌海水中的样品表面附着了大量细胞及其代谢产生的生物膜,并出现了大量微孔与粉化痕迹。通过FTIR结果可以发现,浸泡在假单胞菌海水中的样品的N—H和醚基中的C—O含量明显比无菌海水中浸泡的样品要低。结论 电化学结果表明,假单胞菌显著降低了涂层的腐蚀抗性,并导致了涂层的分解。SEM和FTIR的分析结果证明,假单胞菌通过破坏聚氨酯分子中的N—H和醚基中的C—O,造成了涂层的分解。     

一项微生物能力验证的多种方法检测结果分析

为确保微生物能力验证结果的准确报出,本实验室采用了多种方法进行CNAS T 0710的铜绿假单胞菌(Pseudomonas aeruginosa)和金黄色葡萄球菌(Staphylococcus aureus)的能力验证比对检验,结果表明:以上多种检测方法的测试结果相吻合,均显示5份样品中有2份检出铜绿假单胞菌,2份检出金黄色葡萄球菌。最终5份样品测试均取得满意结果,在分析比较中也总结出各种检验方法的优缺点。     

不同贮藏温度下稻花鸡肉优势腐败菌变化及Arrhenius 货架期预测模型的建立

为获得稻花鸡肉腐败菌的Arrhenius货架期预测模型,采用培养基初步筛选与16S rDNA全基因序列鉴定优势腐败菌,研究不同贮藏温度（25、4、0℃）下优势腐败菌和菌落总数的生长变化,通过化学反应动力学方程构建菌落总数、假单胞菌和沙雷氏菌货架期预测模型并进行验证。结果表明,稻花鸡肉在贮藏过程中逐渐占主导地位的优势腐败菌是假单胞菌属莓实假单胞菌和肠杆菌科沙雷氏菌属液化沙雷氏菌。稻花鸡肉25℃常温贮藏下货架期不超过0.5 d,腐败中后期沙雷氏菌占主导地位,4℃冷藏保鲜货架期不超过4 d,假单胞菌和沙雷氏菌均随贮藏时间的延长呈增长趋势,0℃冰温贮藏货架期不超过10 d,贮藏后期假单胞菌和沙雷氏菌差异性不显著。利用菌落总数、假单胞菌、沙雷氏菌3个指标建立货架期预测模型,3种货架期预测模型预测值与实测值对比,平均相对误差均在允许范围内,预测效果最佳的是假单胞菌货架期预测模型。菌落总数、假单胞菌和沙雷氏菌货架期预测模型均能对稻花鸡肉的货架期进行真实预测。     

Immobilization of lipase onto metal–organic frameworks for enantioselective hydrolysis and transesterification

Enzyme immobilization enhances the catalytic activity and stability of the enzyme, and also improves reusability. Metal–organic frameworks (MOFs), which possess diversified structures and porosity, have been used as excellent carriers for enzyme immobilization. Pseudomonas fluorescens lipase (PFL) has been successfully immobilized onto MOFs by covalent cross-linking to obtain a series of immobilized lipase (PFL@MOFs). PFL@MOFs are used for catalytic enantioselective hydrolysis of 2-(4-hydroxyphenyl) propionic acid ethyl ester enantiomers (2-HPPAEE) in aqueous medium and transesterification of 4-methoxymandelic acid enantiomers (4-MMA) in organic medium. The experimental results indicated that PFL@Uio-66(Zr) exhibits excellent enzymatic catalysis performances and high enantioselectives. In addition, to improve catalytic activity and reusability, PFL is modified by the polyethylene glycol (PEG) to prepare PEG-modified lipase (PFL-PEG), then PFL-PEG is immobilized onto Uio-66(Zr) to prepare PFL-PEG@Uio-66(Zr), demonstrating better reusability and catalytic activity compared with PFL@Uio-66(Zr).     

Benzyl Isothiocyanate Attenuates Inflammasome Activation in Pseudomonas aeruginosa LPS-Stimulated THP-1 Cells and Exerts Regulation through the MAPKs/NF-κB Pathway

Inflammasomes are a group of intracellular multiprotein platforms that play important roles in immune systems. Benzyl isothiocyanate (BITC) is a constituent of cruciferous plants and has been confirmed to exhibit various biological activities. The modulatory effects of BITC on inflammasome-mediated interleukin (IL)-1β expression and its regulatory mechanisms in Pseudomonas aeruginosa (P. aeruginosa) LPS/ATP-stimulated THP-1 cells was investigated. Monocytic THP-1 cells were treated with phorbol myristate acetate (PMA) to induce differentiation into macrophages. Enzyme-linked immunosorbent assays (ELISA) were performed to measure the levels of IL-1β produced in P. aeruginosa LPS/ATP-exposed THP-1 cells. Western blotting was performed to examine the BITC modulatory mechanisms in inflammasome-mediated signaling pathways. BITC inhibited IL-1β production in P. aeruginosa LPS/ATP-induced THP-1 cells. BITC also inhibited activation of leucine-rich repeat protein-3 (NLRP3) and caspase-1 in P. aeruginosa LPS/ATP-induced THP-1 cells. Furthermore, we show that mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) activation in P. aeruginosa LPS was attenuated by BITC. These BITC-mediated modulatory effects on IL-1β production may have therapeutic potential for inflammasome-mediated disorders such as a nasal polyp.     

Modeling of Rhamnolipid Biosurfactant Production: Estimation of Kinetic Parameters by Genetic Algorithm

Studies about kinetics and modeling of production parameters for biosurfactants are essential to the development of efficient processes from an economic point of view. In this sense, this work evaluated the performance of four nonstructured models to explain the experimental data for biomass growth, substrate consumption, and rhamnolipid production using glycerol as carbon source and a Pseudomonas aeruginosa strain. The kinetic parameters of each model were estimated using a global search method known as genetic algorithm and numerical discretization of differential equations by the Runge–Kutta 4th order method. The main result of this study showed that the Monod model best represented the experimental data, with μ_max values of 0.06 h⁻¹, K_S of 50.8 g L⁻¹, Y_X/S of 0.43 g g⁻¹, and Y_P/X equal to 0.017 g g⁻¹.