Microbiological safety of kinema, a fermented soya bean food

Kinema is a fermented soya bean food of Nepal and the hilly regions of North-eastern States of India. Generally, the fermentation is dominated by Bacillus spp. that often cause alkalinity and desirable stickiness in the product. The present study was undertaken in a limited number of commercial (market) kinema samples to test for the presence of foodborne pathogens and their properties. Bacillus cereus was present in numbers exceeding 10⁴ cfu/g product in five of the tested 15 market samples. Enterobacteriaceae and coliform bacteria exceeded 10⁵ cfu/g in 10 of the 15 samples. Escherichia coli exceeding 10⁵ cfu/g was found in two samples. Staphylococcus aureus was not detected in any of the tested samples. Of 31 isolated typical and atypical strains of B. cereus, 18 representative strains were tested qualitatively for the ability to produce diarrhoeal type enterotoxin (BCET) using an Oxoid BCET-RPLA test kit. Overall, BCET was formed by 12 strains in BHIG (brain heart infusion broth +1% glucose), by seven strains on sterilized cooked rice, and by five strains on sterilized cooked soya beans. Semi-quantitative tests on BCET revealed that levels exceeding 256 ng/g soya beans, produced by single pure culture inoculation with the isolated B. cereus strains, were reduced to ≤ 8 ng/g by frying kinema in oil, a common procedure when making kinema curry. It was also shown in a mixed pure culture experiment that a kinema strain B. Subtilis DK-W1, is able to suppress growth and BCET formation by a selected toxin producing strain (BC7-5) of B. cereus. It is concluded that the traditional way of making kinema and its culinary use in curries is safe. However, for novel applications of kinema, safety precautions are advisable.     

The Co-Culture of Staphylococcal Biofilm and Fibroblast Cell Line: The Correlation of Biological Phenomena with Metabolic NMR1 Footprint

Staphylococcus aureus is one of the most prevalent pathogens associated with several types of biofilm-based infections, including infections of chronic wounds. Mature staphylococcal biofilm is extremely hard to eradicate from a wound and displays a high tendency to induce recurring infections. Therefore, in the present study, we aimed to investigate in vitro the interaction between S. aureus biofilm and fibroblast cells searching for metabolites that could be considered as potential biomarkers of critical colonization and infection. Utilizing advanced microscopy and microbiological methods to examine biofilm formation and the staphylococcal infection process, we were able to distinguish 4 phases of biofilm development. The analysis of staphylococcal biofilm influence on the viability of fibroblasts allowed us to pinpoint the moment of critical colonization—12 h post contamination. Based on the obtained model we performed a metabolomics analysis by ¹H NMR spectroscopy to provide new insights into the pathophysiology of infection. We identified a set of metabolites related to the switch to anaerobic metabolism that was characteristic for staphylococcal biofilm co-cultured with fibroblast cells. The data presented in this study may be thus considered a noteworthy but preliminary step in the direction of developing a new, NMR-based tool for rapid diagnosing of infection in a chronic wound.     

Investigation of the Anti-Methicillin-Resistant Staphylococcus aureus Activity of (+)-Tanikolide- and (+)-Malyngolide-Based Analogues Prepared by Asymmetric Synthesis

Herein, we report antibacterial and antifungal evaluation of a series of previously prepared (+)-tanikolide analogues. One analogue, (4S,6S)-4-methyltanikolide, displayed promising anti-methicillin-resistant Staphylococcus aureus activity with a MIC of 12.5 µg/mL. Based on the antimicrobial properties of the structurally related (−)-malyngolide, two further analogues (4S,6S)-4-methylmalyngolide and (4R,6S)-4-methylmalyngolide bearing a shortened n-nonyl alkyl side chain were prepared in the present study using a ZrCl₄-catalysed deprotection/cyclisation as the key step in their asymmetric synthesis. When these were tested for activity against anti-methicillin-resistant Staphylococcus aureus, the MIC increased to 50 µg/mL.     

液体消毒洗涤剂的研制

研制了具有消毒杀菌、洗涤去污双重功效的液体消毒洗涤剂 ,测定了其对金黄色葡萄球菌的杀灭效果。结果表明 ,消毒洗涤剂的杀菌效果随其浓度的增加 ,作用时间的延长而提高 ;戊二醛浓度为 2 0 0× 10 -6时 ,与金黄色葡萄球菌作用 5min ,杀灭率可达 99.99%。将消毒洗涤剂在室温下放置半年 ,戊二醛分解率为 17.3 1%     

Extranasal Staphylococcus aureus colonization predisposes to bloodstream infections in patients on hemodialysis with noncuffed internal jugular vein catheters

Introduction: Staphylococcal infection of endogenous origin is an important cause of morbidity and mortality in patients who receive hemodialysis (HD). The risk of such infections in nasal carriers of the organism is well defined. Extranasal carriage of the organism at extranasal sites may pose similar risks. Methods: A total of 70 patients about to undergo internal jugular vein catheterization for HD were enrolled in this prospective observational study. Swab cultures were obtained from anterior nares, posterior pharynx, axillae, toe web spaces, and vascular access sites at baseline and 1 week later. A patient was defined as a persistent carrier when the same organism was grown in both samples. Staphylococcus aureus bloodstream infections were assessed by blood and catheter tip cultures over a 90‐day period. Findings: The mean age of the patients was 43.71 ± 16.2 years. Persistent S. aureus carriage at anterior nares, throat, axilla, toe web spaces, vascular access site, and all sites was documented in 27.9%, 11.4%, 40%, 32.9%, 4.3%, and 64.2% of patients, respectively. Fifteen patients developed S. aureus infections. Catheter related S. aureus infections (CRI) were more likely in persistent carriers than nonpersistent carriers with odds ratios (95% CI) of 10.2 (2.8–37.1), 8.6 (1.7–42.2), 17.3 (3.4–86.0), 3.0 (0.9–9.8), and 1.9 (0.2–22.4) for anterior nares, throat, axilla, toe web spaces, and vascular access site carriers, respectively. The probability of developing CRI in persistent S. aureus carriers was 55% compared to none in noncarriers at 90 days (P = 0.04). Discussion: Extranasal S. aureus carriage is as significant a risk factor as nasal carriage for staphylococcal infections in patients on HD through catheters. The study is limited by lack of molecular phenotyping.     

不同混纺比甲壳素／长绒棉混纺织物抗菌性的测试与分析

为了找出抗菌性好又经济实用的甲壳素／长绒棉混纺织物的混纺比,采用琼脂平皿扩散法（定性）和吸收法（定量）对5种不同混纺比的甲壳素／长绒棉混纺织物进行抗菌性测试,突出抗菌性能测试方法的改进——表面活性剂的选择、接种菌液量以及涂板培养时间．实验结果表明,混纺织物对金黄色葡萄球菌的抑菌率大于大肠杆菌,而且只要甲壳素的混纺比例达到20％,对金黄色葡萄球菌的抑菌率能达到99．88％,对大肠杆菌的抑菌率能达到91．2％,再提高甲壳素的比例则无法取得更好的性价比．     

不同制备条件对SpA分子印迹聚合微球聚合效果的影响

本文用金黄色葡萄球菌蛋白A(SpA)作为模板,以丙烯酰胺为功能单体,利用反相悬浮聚合法合成了SpA分子印迹聚合微球.通过扫描电镜观测不同条件下聚合微球的聚合效果,研究不同制备条件对SpA分子印迹聚合微球聚合效果的影响,从而筛选制备聚合微球最佳工艺;并检测了制备的分子印迹聚合物对SpA的吸附性能.结果表明,反相悬浮聚合法合成SpA分子印迹微球的最佳工艺条件为:分散剂乙基纤维素(EC)加入量0.20 g/100 mL(甲苯);甲苯与水的比例10:3;交联度10%;引发剂加入方式为缓慢滴加;反应温度50℃.反相悬浮聚合法制备的SpA分子印迹聚合微球对SpA的吸附量:6.956×10-3μmol/g.     

源自螺旋藻渣枯草芽孢杆菌发酵抗菌肽SP-AP-1和伊枯草菌素对金黄色葡萄球菌抑菌机制对比研究

目的:研究枯草芽孢杆菌发酵螺旋藻渣的发酵液中分离出的抗菌肽SP-AP-1和伊枯草菌素对金黄色葡萄球菌的抑菌机理.方法:采用紫外分光光度法、邻硝基苯-β-D-吡喃半乳糖苷法和原子吸收光谱法研究抗菌肽对金黄色葡萄球菌细胞膜通透性的影响;采用考马斯亮蓝法、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和琼脂糖凝胶电泳考察抗菌肽对金黄色葡...     

Bioconjugated Thymol-Zinc Oxide Nanocomposite as a Selective and Biocompatible Antibacterial Agent against Staphylococcus Species

Owing to the rapid spread of antibiotic resistance among Staphylococcus species, effective and low-risk alternatives to antibiotics are being actively searched. Thymol (THO), the most abundant component of the oil extracted from thyme, can be considered as a natural antibacterial alternative. However, the low antibacterial activity and non-selectivity of THO limit its usage as a universal anti-Staphylococcus agent. Herein, we report the bioconjugation of THO with ZnO nanoparticle (ZO), which resulted in the TZ nanocomposite (NC), as a potent and selective antibacterial agent against Staphylococcus species, particularly S. epidermidis. The cell-free supernatant (CFS) of ATCC 25923 cultures was employed for the production of TZ NC. Successful production of TZ NC was confirmed via X-ray diffraction (XRD), Fourier-transform infrared (FT-IR) spectroscopy, and ultraviolet–visible (UV–Vis) studies. TZ NC had selective efficacy against Staphylococcus species, with MIC values 2–32-fold lower than THO. The antibacterial mechanisms of TZ NC are proposed to involve membrane rupture, suppression of biofilm formation, and modulation of new cell wall and protein-synthesis-associated cellular pathways. Its biocompatibility against HCT116 cells was also checked. Our findings suggest that the TZ nanocomposite could improve the selectivity and bactericidal activity of THO against target species.     

Evaluation of the Effects of Ag,Cu, ZnO and TiO2 Nanoparticles on the Expression Level of Oxidative Stress-Related Genes and the Activity of Antioxidant Enzymes in Escherichia coli,Bacillus cereus and Staphylococcus epidermidis

Although the molecular response of bacteria exposed to metal nanoparticles (NPs) is intensively studied, many phenomena related to their survival, metal uptake, gene expression and protein production are not fully understood. Therefore, this work aimed to study Ag-NPs, Cu-NPs, ZnO-NPs and TiO₂-NPs-induced alterations in the expression level of selected oxidative stress-related genes in connection with the activity of antioxidant enzymes: catalase (CAT), peroxidase (PER) and superoxide dismutase (SOD) in Escherichia coli, Bacillus cereus and Staphylococcus epidermidis. The methodology used included: the extraction of total RNA and cDNA synthesis, the preparation of primers for selected housekeeping and oxidative stress genes, RT-qPCR reaction and the measurements of CAT, PER and SOD activities. It was established that the treatment of E. coli and S. epidermidis with NPs resulted mainly in the down-regulation of targeted genes, whilst the up-regulation of genes was confirmed in B. cereus. The greatest differences in the relative expression levels of tested genes occurred in B. cereus and S. epidermidis treated with TiO₂-NPs, while in E. coli, they were observed under ZnO-NPs exposure. The changes found were mostly related to the expression of genes encoding proteins with PER and CAT-like activity. Among NPs, ZnO-NPs and Cu-NPs increased the activity of antioxidants in E. coli and B. cereus. In turn, TiO₂-NPs had a major effect on enzymes activity in S. epidermidis. Considering all of the collected results for tested bacteria, it can be emphasised that the impact of NPs on the antioxidant system functioning was dependent on their type and concentration.