Growth, survival and inactivation of Pseudomonas aeruginosa and Staphylococcus aureus strains of various origin in the presence of ethanol

The influence of ethanol on the behaviour of Pseudomonas aeruginosa and Staphylococcus aureus strains was evaluated throughout this study. Strains of different origin were used: collection, clinical and industrial strains were selected. Concentrations of ethanol from 0 to 20% (v/v) were evaluated by automated optical density measurements and by enumeration. When growth conditions were observed, predictive microbiology models were used to assess quantitatively for the ethanol effect. Primary modelling of kinetics was performed to determine growth rate values; secondary modelling was performed on these growth rates as influenced by ethanol, and minimum inhibitory concentrations of ethanol were determined for each strain. Staphylococcus aureus strains were more resistant to ethanol than P. aeruginosa strains, in growth conditions as well as in inactivation conditions. Furthermore, clinical S. aureus strains were more resistant than the collection strain. The method was promising for management of microbiological safety in cosmetics.     

低温等离子体对金黄色葡萄球菌和铜绿假单胞菌的杀菌效果及动力学特性

为研究介质阻挡放电低温等离子体对金黄色葡萄球菌和铜绿假单胞菌的杀菌作用,以菌悬液（1×108 CFU/mL）为研究对象,分析其杀菌效果,通过Linear、Weibull和Log-linear+Shoulder+Tail三种数学模型,分析低温等离子体杀菌动力学特性,以均方根误差（RMSE）、决定系数(R2)、准确度因素(Af)和偏移因素(Bf)四个参数作为评价模型拟合度优劣的指标。实验结果表明,当O2:N2=65:35,介质厚度为2 mm,处理电压为75 kV,处理时间为150 s,处理后放置0 h时,金黄色葡萄球菌和铜绿假单胞菌的杀菌率分别为90.00%和98.99%;当处理后放置时间延长至2 h时,二者杀菌率提高至90.27%和99.61%。拟合结果表明,Weibull模型和Log-linear+Shoulder+Tail模型比Linear模型更符合低温等离子体处理对金黄色葡萄球菌和铜绿假单胞菌的实际杀菌曲线,而Log-linear+Shoulder+Tail模型适用面更广,可以更好的描述不同处理后放置时间下的低温等离子体的杀菌过程。     

鹿蹄草素对金黄色葡萄球菌和绿脓杆菌的抑制效果观察

对鹿蹄草素作用下金黄色葡萄球菌和绿脓杆菌生长曲线、膜通透性和扫描电镜下形态进行研究,初步探讨了鹿蹄草素抑菌和杀菌机理.结果表明:鹿蹄草素能有效抑制金黄色葡萄球菌和绿脓杆菌的生长,抑制作用随着药物浓度增加而增加,且对金黄色葡萄球菌具有杀灭作用.鹿蹄草素作用细菌后,金黄色葡萄球菌细胞膜通透性与空白对照组相比显著提高,绿脓杆菌无明显变化.扫描电镜结果表明,鹿蹄草素作用后金黄色葡萄球菌菌体细胞结构被明显破坏,细胞壁呈溶解状,菌体细胞间相互粘结,细胞间界限变模糊;绿脓杆菌菌体细胞间界限变模糊,但并未对菌体细胞壁和细胞膜造成破坏.结果分析认为,鹿蹄草素对金黄色葡萄球菌的抑制和杀灭与菌体细胞膜和细胞壁结构被破坏直接相关,而鹿蹄草素对绿脓杆菌的抑制与菌体细胞膜和细胞壁无关.     

国内外包装饮用水标准中微生物指标比较和铜绿假单胞菌指标建议

近年来,国内包装饮用水卫生状况的检测中微生物卫生指标不合格情况较为严重,而饮用水的卫生情况直接关系到民大的身体健康。根据GB 19298-2014《食品安全国家标准包装饮用水》,并参照世界卫生组织《饮用水水质指南》第四版,本文对我国的包装饮用水相应的微生物指标进行分析比较,指出存在的问题并提出修改的建议,为包装饮用水的安全生产提供参考。     

Adhesion and detachment kinetics of several strains of Staphylococcus aureus subsp. aureus under three different experimental conditions

The kinetics of adhesion of five Staphylococcus aureus subsp. aureus strains (CECT 976, 4459, 4465, 4466 and 5191) to polypropylene at 25 degrees C in the absence of nutrients (PBS medium) were initially compared. Those strains with the highest (CECT 4459) and the lowest (CECT 976) adhesion levels were selected for further studying the effects of a nutrient-rich adhesion-promoting medium (TSB plus 1% glucose-TSBG) as well as of a conditioning film consisting of dried mussel cooking juices (MCJ) on adhesion to and detachment from polypropylene surfaces. Adhesion kinetics were properly described by an empirical model in the absence of conditioning film. The maximum adhesion level was much higher in the presence of TSBG than in PBS, decreasing sharply in both cases after 10-15 h. In contrast, adhesion increased exponentially during 25 h in the presence of dried MCJ. Clear differences were thus found in different media, and it suggests that cleaning strategies should vary under different conditions. The comparison of the adhesion strengths under the different experimental conditions showed that the persistence was highest when biofilms were formed on MCJ, which indicates that cells would remain longer as a source of cross-contamination. Some biofilms were examined by electronic microscopy, and different structures were observed under the different experimental conditions. It is concluded that the study of biofilm formation by S. aureus is necessary to establish efficient control systems in the food industry.     

加压CO2对污染乳中金黄色葡萄球菌杀菌效果的研究

通过加压二氧化碳对人工污染金黄色葡萄球菌的乳进行处理,对时间,温度,压力三个单因素试验对金黄色葡萄球菌的杀菌情况进行研究。结果显示单因素试验中,伴随着因素水平的增加,杀菌率随之提高。然后选取三个因素中最适宜的条件进行正交试验,并通过方差分析得出压力因素对金黄色葡萄球菌杀菌效果最显著、温度次之,时间不显著。在7.5 MPa,45℃,60 min时,杀菌效果最好达到98%以上。     

Effect of microbial sanitizers for reducing biofilm formation of Staphylococcus aureus and Pseudomonas aeruginosa on stainless steel by cultivation with UHT milk

Food Science and Biotechnology - Biofilm is a serious issue in the dairy factory due to it increases the opportunity for microbial contamination. Staphylococcus aureus and Pseudomonas aeruginosa...     

Bactericidal and antibiofilm properties of Rumex japonicus Houtt. on multidrug-resistant Staphylococcus aureus isolated from milk

Multidrug-resistant (MDR) Staphylococcus aureus and its biofilm formation have been challenging to control in milk and dairy industries. Biofilms formed by Staph. aureus may result in the failure of antibacterial agents and disinfectants to penetrate the biofilm in an attempt to control contamination. Novel natural antibacterial agents are required to combat MDR bacteria and biofilms. In this study, we evaluated the bactericidal, antibiofilm, and antimotility effects of Rumex japonicus Houtt. (RJH) extract on MDR Staph. aureus isolated from milk. The RJH extract exhibited good antibacterial activity against MDR strains with minimum inhibitory concentrations (MIC) ranging from 0.78 to 6.25 mg/mL and minimum bactericidal concentrations ranging from 3.125 to 12.5 mg/mL. The extract showed strong inhibition of biofilm formation (81.9%) at sub-MIC value and eradication of biofilm at higher concentrations. The motility of Staph. aureus was effectively blocked by the extract. Major compounds emodin, chrysophanol, and physcion were identified in RJH extract using HPLC-linear trap quadrupole (LTQ)/Orbitrap-mass spectrometry. The extract was nontoxic to human epithelial cell lines such as Caco-2 and HT-29 cell lines at concentrations ranging from 0.1 to 0.5 mg/mL, and from 0.1 to 0.75 mg/mL, respectively. These findings suggest that RJH extract could be an alternative to synthetic preservatives in milk and dairy products.     

补加氨水对薛氏丙酸杆菌分批发酵及代谢物抑菌活性的影响

研究了在分批发酵过程中补加氨水对1株薛氏丙酸杆菌(Propionibacterium shermanii)发酵及其代谢物对恶臭假单胞菌(Pseudomonas pudia)抑菌活性的影响。结果表明,薛氏丙酸杆菌代谢物在初始pH7.0时抑菌活性最高,为7.59AU/mL,显著高于其他实验组(p<0.05);每隔24h补加10%氨水溶液调节发酵液pH6.0时,薛氏丙酸杆菌的生物量和代谢物的抑菌活性都最大,分别为9.62mg/mL和16.42AU/mL;研究分别从第2、3或4d补加10%氨水开始调节发酵液pH6.0时发现,从发酵第2d开始调节pH效果最好,生物量和代谢物的抑菌活性分别达到9.55mg/mL和16.47AU/mL。这些结果表明,在分批发酵过程中补加氨水调节发酵液的pH可以显著提高丙酸杆菌代谢物的抑菌活性。      

食源性金黄色葡萄球菌肠毒素基因型分布研究

目的 采用PCR法扩增食源性金黄色葡萄球菌中肠毒素基因以了解该菌肠毒素基因携带情况,比较食物中毒和食品监测来源菌株中肠毒素基因检出率差异.方法 合成sea、seb、sec、sed和see五种肠毒素基因特异性引物,用常规PCR方法扩增食物中毒和食品监测来源菌株中各自肠毒素基因,同时采用mini-VIDAS检测食物中毒来源菌株中肠毒素.结果 110株菌株中有30株检出肠毒素基因,检出率为27.3％,肠毒素基因阳性菌株均只检出1种肠毒素基因.其中来自2起食物中毒的14株菌株均检出seb型肠毒素和相关基因,检出率为100％.来源于食品监测样本的96株菌株中有16株检出肠毒素基因,检出率为16.7％,包括sea型4株、seb型2株、sec型4株、sed型6株.结论 在宁波市食品监测中所分离的金黄色葡萄球菌所携带的肠毒素基因主要有sea、seb、sec和sed四型,而seb型肠毒素是引起金黄色葡萄球菌肠毒素所致食物中毒的主要因素.     

In vitro deposition of lysozyme on etafilcon A and balafilcon A hydrogel contact lenses: effects on adhesion and survival of Pseudomonas aeruginosa and Staphylococcus aureus.

PURPOSE: To compare lysozyme adsorption and absorption and bacterial adhesion interactions on conventional (etafilcon A) and silicone (balafilcon A) hydrogel contact lenses. METHOD: Lysozyme concentrations and activities associated with the lenses were determined after solvent extraction (trifluoroacetic acid/acetonitrile) and directly on the lenses without extraction with micrococcal- and micro-bicinchoninic acid (BCA) assays. Cells of bacteria with radiolabeled leucine and a cell recovery procedure were used in determinations of bacterial adhesion to lenses. RESULTS: Lysozyme was adsorbed and absorbed to the conventional etafilcon A lens at about a 10-fold greater concentration than to the balafilcon A silicone hydrogel lens. Enzyme activities on the surfaces of both lenses were similar but replenished after saline extraction only with the etafilcon A lens. Lysozyme on the lens surface showed significant lysis of Micrococcus luteus but had a negligible effect on the adhesion and survival of Staphylococcus aureus. Lysozyme did not appear to affect the survival of Pseudomonas aeruginosa on lenses. CONCLUSION: In vitro experiments show that concentrations of active lysozyme on the surface of the etafilcon A lens, unlike the balafilcon A lens which showed negligible absorption, may be sustained from the lens matrix. Lysozyme deposited on hydrogel lenses had marked activity against M. luteus but relatively minor effects on the primary adhesion of P. aeruginosa and S. aureus.     

Comparing predicting models for heat inactivation of Listeria monocytogenes and Pseudomonas aeruginosa at different pH

Under the same experimental conditions it has been demonstrated that whereas survival curves of Listeria monocytogenes in the range of temperatures from 54 to 62 °C followed a first-order kinetic, those of Pseudomonas aeruginosa in the range of temperatures from 50 to 56 °C were not linear showing a shoulder followed by a linear region. The first order kinetic model did not describe survival curves of P. aeruginosa. A model based on the Weibull distribution (Log₁₀(N_t/N₀)=(1/−2.303)*(t/b)ⁿ)) accurately described the inactivation kinetics of both microorganisms at the three pHs of 4, 5.5, 7.4 investigated. For both microorganisms, the b value depended on the treatment temperature and the pH of the treatment medium. Whereas for L. monocytogenes the n value was independent of the treatment conditions, for P. aeruginosa the n value depended on the pH of the treatment medium.

The model based on the Weibull distribution was capable of accurately predicting the treatment time to inactivate five Log₁₀ cycles of both microorganisms at the three pHs investigated.     

异养硝化-好氧反硝化菌株的分离、鉴定及其氨氮降解过程初探

该研究采用选择性培养基从汾河底泥中分离筛选异养硝化-好氧反硝化菌,通过形态观察、生理生化试验及分子生物学技术对其进行菌种鉴定,并对其氨氮（NH4+-N）降解过程进行动态监测,测定NH4+-N降解过程中关键酶的基因及酶活。结果表明,分离筛选得到一株异养硝化-好氧反硝化菌,编号为ZH-2,其被鉴定为醋酸钙不动杆菌（Acinetobacter calcoaceticus）。该菌在NH4+-N降解过程中存在NO3--N和NO2--N两种中心代谢产物,其基因组中含有3种关键酶（氨单加氧酶（Amo）、硝酸盐还原酶（Nap）和亚硝酸盐还原酶（Nir））基因,在NH4+-N降解过程,这3种关键酶的比酶活分别为45.69 mU/mg、3.54 mU/mg和6.91 mU/mg。综上,初步确定A. calcoaceticus ZH-2的NH4+-N降解过程是NH4+-N→NH2OH→NO2--N→NO3--N→NO2--N→NO→N2O→N2的经典降解途径,为该菌的进一步推广应用提供了理论依据。     

Correction to: Effect of microbial sanitizers for reducing biofilm formation of Staphylococcus aureus and Pseudomonas aeruginosa on stainless steel by cultivation with UHT milk

Food Science and Biotechnology - The article “Effect of microbial sanitizers for reducing biofilm formation of Staphylococcus aureus and Pseudomonas aeruginosa on stainless steel by...     

金黄色葡萄球菌耐药性与生物被膜能力的鉴定

本文选择常见的典型食源性微生物金黄色葡萄球菌,从食品微生物安全角度出发,对127株葡萄球菌的菌株特性、耐药性与生物被膜生长能力进行研究,包括菌株生化鉴定;PCR扩增葡萄球菌特异16S rRNA与金葡菌特异femA基因,通过对耐药基因mecA和orfX检测确定菌株的耐药特性;最后,运用结晶紫染色法进行生物被膜能力检测。127株菌株包括119株金葡菌(均携带16S rRNA与femA)与8株凝固酶阴性葡萄球菌8株(仅携带16S rRNA)。119株金葡菌中,107株携带mecA基因与orfX基因,为耐甲氧西林金葡球菌(MRSA);8株凝固酶阴性葡萄球菌中,均携带mecA基因。所有菌株均能生成生物被膜,其中能形成强、中等与弱粘附生物被膜能力菌株分别有5(3.9%)、47(37.0%)和75(59.1%)株。金葡菌中耐药性与生物被膜较为普遍,由于食源性微生物形成生物被膜后,具有逃逸常规消毒和杀菌手段的能力,成为食品安全中的潜在隐患。     

Efficacy of neutral electrolyzed water to inactivate Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus on plastic and wooden kitchen cutting boards

This study evaluated the efficacy of neutral electrolyzed water (NEW; 64.1 mg/liter of active chlorine) to reduce populations of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Listeria monocytogenes on plastic and wooden kitchen cutting boards. Its effectiveness was compared with that of a sodium hypochlorite solution (NaClO; 62.3 mg/liter of active chlorine). Inoculated portions of cutting boards were rinsed in either NEW or NaClO solutions, or deionized water (control). Plastic boards were rinsed for 1 min and wooden boards for 1 and 5 min. After each treatment, the surviving population of each strain was determined on the surface and in the soaking water. No significant difference (P > or = 0.05) was found between the final populations of each strain with regard to the treatment solutions (NEW or NaClO). However, a significant difference (P < or = 0.05) was revealed between surface materials after 1 min of washing. Whereas in plastic boards the initial bacterial populations were reduced by 5 log CFU/50 cm2, in wooden cutting boards they underwent a reduction of <3 log CFU/50 cm2. A 5-min exposure time yielded reductions of about 4 log CFU/50 cm2. The surviving populations of all bacteria in NEW and NaCIO washing solutions were <1 log CFU/ml after soaking both surfaces. This study revealed that NEW treatment is an effective method for reducing microbial contamination on plastic and wooden cutting boards. NEW efficacy was comparable to that of NaCIO, with the advantage of having a larger storage time.     

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。     

安石榴苷的纯化及对金黄色葡萄球菌抑菌作用的研究

采用超声波辅助法从石榴皮中提取单宁类物质,并用Amberlite XAD-16大孔吸附树脂进行柱层析,以甲醇梯度洗脱法进行纯化,经高效液相色谱对其组分进行分析后,以安石榴苷含量最高的组分对金黄色葡萄球菌进行抑菌实验,分别测得抑菌圈直径和最小抑菌浓度,并检测了安石榴苷对金黄色葡萄球菌生长曲线的影响。液相结果表明,随着洗脱剂浓度的升高,安石榴苷的含量呈现先升高后降低的趋势,最高含量达79.6%;抑菌结果表明,安石榴苷纯化物对金黄色葡萄球菌有明显的抑菌效果,最小抑菌浓度为0.1mg/m L,安石榴苷纯化物以剂量依赖的形式抑制金黄色葡萄球菌的生长。      

1874株金黄色葡萄球菌临床分布及耐药性结果分析

目的了解四川省某医院甲氧西林敏感金黄色葡萄球菌(methicillin sensitive Staphylococcus aureus,MSSA)与耐甲氧西林金黄色葡萄球菌(methicillin resistant Staphylococcus aureus,MRSA)的临床科室分布和耐药性情况。方法回顾性分析2014年12月-2017年12月四川省某医院门诊及住院患者,包括8名因金黄色葡萄球菌引起的食物中毒患者,送检各类标本分离出SA(Staphylococcus aureus)可疑菌落1874株,对其进行鉴定和药敏检测。结果 MSSA和MRSA标本主要来源于痰液(51.31%,74.33%)、伤口分泌物(21.52%,8.96%)和脓液(10.07%,7.92%)。725株MSSA临床分离率依次为门诊(27.17%)、呼吸科(15.45%)、ICU(10.21%)、神经科(8.55%)和口腔科(5.52%),1149株MRSA依次为ICU(33.16%)、神经科(11.66%)、呼吸科(9.75%)、儿科(7.57%)和妇产科(6.27%)。耐药性结果表明,MSSA对苯唑西林、万古霉素、利奈唑胺、替考拉林敏感,对青霉素G、红霉素和阿莫西林/克拉维酸耐药率依次为89.38%、56.69%、21.51%外,对其他抗菌药耐药率均18.00%; MRSA对万古霉素利、奈唑胺、替考拉林敏感,对苯唑西林、莫西沙星、克林霉素、红霉素、青霉素G、四环素、环丙沙星、阿莫西林/克拉维酸和氨苄西林/舒巴坦的耐药率均显著高于MSSA。结论四川省某医院2014年12月-2017年12月期间临床分离的SA以MRSA为主, MSSA和MRSA在标本来源、科室分布等方面存在较大差异, MRSA耐药情况更为严重,明显高于MSSA。若要评估金黄色葡萄球菌感染风险以及病人的治疗方案,应分别考虑MRSA和MSSA的感染来源和耐药性。