表皮葡萄球菌耐药性与生物被膜的相关性

目的分析表皮葡萄球菌耐药性与生物被膜的相关性。方法采用刚果红平板法检测表皮葡萄球菌胞间多糖黏附素(PIA);半定量生物被膜形成试验检测生物被膜表型;采用二倍稀释法检测各药物对该菌的最低抑菌浓度(MIC)。结果42株表皮葡萄球菌中,9株为PIA+/生物被膜表型+,4株为PIA+/生物被膜表型-,29株为PIA-/生物被膜表型-;3组菌对苯唑西林、克林霉素、阿米卡星、左氧氟沙星和阿莫西林/克林霉素的耐药性差异有统计学意义;万古霉素和阿莫西林/克拉维酸对3组菌的MIC低于NCCLS给出的敏感值,青霉素、苯唑西林、克林霉素、红霉素和头孢西丁对3组菌的MIC则远远高于NCCLS给出的敏感值,且β-内酰胺类的3种药物对3组菌的MIC差异有统计学意义。结论表皮葡萄球菌生物被膜形成是多基因调控过程,在细菌耐药中具有重要作用。     

硝酸改性污泥基生物炭除U(Ⅵ)效果及机理分析

以城市污泥为原料制备出污泥基生物炭,并通过硝酸改性得到硝酸改性污泥基生物炭（SSB-AO）,探究了SSB-AO投加量、溶液初始pH、离子强度、吸附时间、U(Ⅵ)初始质量浓度以及吸附温度等对SSB-AO去除U(Ⅵ)的影响,通过SEM-EDS、FTIR及XPS分析SSB-AO对U(Ⅵ)的去除机理。结果表明：SSB-AO对U(Ⅵ)的吸附符合拟二级动力学模型,吸附过程以化学吸附为主;等温吸附过程符合Langmuir模型。在30 ℃、NaNO3浓度为0.01 mol/L、吸附时间300 min、初始pH=6、U(Ⅵ)初始质量浓度为10~100 mg/L及SSB-AO投加量为0.6 g/L的条件下,SSB-AO去除U(Ⅵ)的理论最大吸附量为80.34 mg/g;通过5次吸附-解吸实验,其吸附率保持在88%以上,说明SSB-AO具有良好的重复使用性;SSB-AO去除U(Ⅵ)的机理为内表面络合作用、静电作用以及离子交换。研究显示硝酸处理污泥基生物炭能有效地提高其对U(Ⅵ)的吸附能力,为含U(Ⅵ)废水处理提供借鉴。     

邻、间、对双胍基苯甲酸盐酸盐抗菌活性的研究

以邻、间、对氨基苯甲酸为起始原料,在盐酸存在下分别与双氰胺合成了中间体邻、间、对双胍基苯甲酸盐酸盐,分别研究了邻、间、对双胍基苯甲酸盐酸盐对革兰氏菌的抗菌活性。实验表明,邻、间、对双胍基苯甲酸盐酸盐对大肠杆菌、金黄色葡萄球菌都具有抑制作用,抗菌活性随浓度的增加而增强;邻双胍基苯甲酸盐酸盐对大肠杆菌、金黄色葡萄球菌最低抑菌浓度分别为0.064mg.mL-1和0.032mg.mL-1,优于间、对双胍基苯甲酸盐酸盐。     

13种杀菌剂对生姜腐霉茎基腐病菌的毒力测定

[目的]通过13种杀菌剂的室内毒力测定,筛选出对生姜茎基腐病有显著抑菌效果的杀菌剂.[结果]供试杀菌剂对生姜腐霉菌——群结腐霉(Pythium myriotylum)菌丝生长的抑制作用存在明显差异,98％(噁)霉灵可湿性粉剂和60％吡唑醚菌酯·代森联水分散性粒剂对生姜腐霉菌——群结腐霉的菌丝生长有显著抑制效果,其抑制中质量浓度EC50值分别为1.9518、2.8952mg/L.25％啶菌(噁)唑(SYP-Z048)乳油、25％吡唑醚菌酯乳油和20％溴硝醇可湿性粉剂的抑制效果也较好,其EC50值分别为4.2275、4.7926、11.6324mg/L.[结论](噁)霉灵、吡唑醚菌酯·代森联、啶菌(噁)唑抑菌效果显著,可作为防治该病的首选药剂.     

表皮葡萄球菌与血管内皮细胞的黏附作用及其机制

目的探讨表皮葡萄球菌与人脐静脉血管内皮细胞ECV304的黏附作用及其机制。方法将表皮葡萄球菌临床分离株通过刚果红琼脂检测胞间多糖黏附素(PIA);半定量生物被膜形成试验检测生物被膜表型;并在体外与ECV304细胞共同作用,在倒置显微镜下观察表皮葡萄球菌对ECV304细胞的黏附数。结果表皮葡萄球菌黏附ECV304细胞的数量随作用时间的延长而增加;PIA+/生物被膜表型+菌株与ECV304细胞的黏附数多于PIA+/生物被膜表型-菌株和PIA-/生物被膜表型-菌株,且差异有统计学意义,而PIA+/生物被膜表型-菌株和PIA-/生物被膜表型-菌株与细胞的黏附数差异无统计学意义。结论表皮葡萄球菌与血管内皮细胞的黏附有时间依赖性,能形成生物被膜的表皮葡萄球菌更易与血管内皮细胞黏附。     

固定化微生物对近海河道黑臭水体的净化研究

考察固定化复合菌对近海河道黑臭水体中COD_(Cr)、NH3-N、TN、TP的去除效果。结果表明:常温条件下,在黑臭水体pH值为7.01,溶解氧的质量浓度为2~3 mg/L,固定化复合菌投加量为6~9 g/L,反应时间为72 h时,净化效果较好,COD_(Cr)的质量浓度由初始100.00降至44.00 mg/L,去除率为56%;NH3-N的质量浓度由初始0.87降至0.03 mg/L,去除率为97%;TN的质量浓度由初始2.50降至1.00 mg/L,去除率为60%;TP的质量浓度由初始2.42降至1.01 mg/L,去除率为58%。相同试验条件下,对比了固定化复合菌、粉状菌、液态菌的处理效果,结果表明:3种菌剂去除效果按照由好变差的排序依次为固定化复合菌、液态菌、粉状菌,其中固定化复合菌的整体成本最低。     

煤矸石基多孔材料对废水中Pb2+的吸附研究

以煤矸石为主要原料,采用发泡法制备煤矸石基多孔材料,并将其作为吸附剂用于对废水中Pb2+的吸附,研究了多孔材料的煅烧温度,多孔材料的添加量、吸附时间、Pb2+初始浓度等条件对Pb2+吸附效果的影响,研究结果表明:在反应温度为25℃,多孔材料投加量0.8 g,Pb2+初始浓度为100 mg/L,吸附时间为10 min时,煤矸石基多孔材料对Pb2+的最佳吸附率为99.92％,吸附量为12.48 mg/L.     

双二茂铁基希夫碱的合成及抗菌活性

利用二茂铁甲醛分别与1,2-乙二胺、1,3-丙二胺和1,4-丁二胺缩合,合成了3种双二茂铁基希夫碱,进行了生物活性实验。结果表明,3种双二茂铁基希夫碱对大肠埃希氏菌和金黄色葡萄球菌均具有较好的抗菌活性,1,3-丙二胺双缩二茂铁甲醛对金黄色葡萄球菌的抑菌能力最强,生物效价为210 U/mg（与四环素盐酸盐相比较）;1,4-丁二胺双缩二茂铁甲醛对大肠埃希氏菌的抑菌能力最好,生物效价为1 058 U/mg（与四环素盐酸盐相比较）。3种双二茂铁基希夫碱对大肠埃希氏菌的抑菌能力比对金黄色葡萄球菌的抑菌能力强。     

偕胺肟基乙烷桥键介孔二氧化硅对铀的吸附研究

采用水热法合成了氰基化乙烷桥键的介孔材料,经胺肟功能化后制备出偕胺肟基介孔SiO_2材料(AO-PMOs)。利用AO-PMOs对铀溶液进行吸附实验,探讨了铀溶液初始质量浓度、pH值以及吸附时间对AO-PMOs吸附铀的影响,并采用FTIR,SEM,EDS技术进行表征分析。结果表明:在溶液初始pH值为5,铀质量浓度为10 mg/L,吸附剂投加量为20 mg,反应温度为303 K的条件下,吸附效果较佳,吸附率最大为99.04%,吸附量可达25 mg/g。该吸附过程与准二级动力学模型(R~20.99),Langmuir和Freundlich等温吸附模型(R~20.98)能较好地拟合。偕胺肟基在AO-PMOs吸附铀过程起主要作用。     

沼泽红假单胞菌对Zn^2＋的吸附性能研究

采用沼泽红假单胞菌为生物吸附剂,研究了锌离子初始浓度、pH值、吸附剂用量、菌龄、吸附时间等因素对沼泽红假单胞菌生物吸附模拟废水中锌的影响,探讨了吸附动力学特征。结果表明,沼泽红假单胞菌在Zn2＋初始质量浓度为200 mg/L、吸附剂用量0.5 g/L、菌龄32 h、pH=7、吸附时间80 min的条件下,对Zn2＋的吸附量为88.23 mg/g;其吸附行为可以用Langmuir和Freundlich吸附等温方程描述,但更符合Freundlich吸附等温方程。对Zn2＋的吸附过程符合二级动力学特征。     

Low Concentration of the Neutrophil Proteases Cathepsin G,Cathepsin B,Proteinase-3 and Metalloproteinase-9 Induce Biofilm Formation in Non-Biofilm-Forming Staphylococcus epidermidis Isolates

Neutrophils play a crucial role in eliminating bacteria that invade the human body; however, cathepsin G can induce biofilm formation in a non-biofilm-forming Staphylococcus epidermidis 1457 strain, suggesting that neutrophil proteases may be involved in biofilm formation. Cathepsin G, cathepsin B, proteinase-3, and metalloproteinase-9 (MMP-9) from neutrophils were tested on the biofilm induction in commensal (skin isolated) and clinical non-biofilm-forming S. epidermidis isolates. From 81 isolates, 53 (74%) were aap⁺, icaA⁻, icaD⁻ genotype, and without the capacity of biofilm formation under conditions of 1% glucose, 4% ethanol or 4% NaCl, but these 53 non-biofilm-forming isolates induced biofilm by the use of different neutrophil proteases. Of these, 62.3% induced biofilm with proteinase-3, 15% with cathepsin G, 10% with cathepsin B and 5% with MMP -9, where most of the protease-induced biofilm isolates were commensal strains (skin). In the biofilm formation kinetics analysis, the addition of phenylmethylsulfonyl fluoride (PMSF; a proteinase-3 inhibitor) showed that proteinase-3 participates in the cell aggregation stage of biofilm formation. A biofilm induced with proteinase-3 and DNAse-treated significantly reduced biofilm formation at an early time (initial adhesion stage of biofilm formation) compared to untreated proteinase-3-induced biofilm (p < 0.05). A catheter inoculated with a commensal (skin) non-biofilm-forming S. epidermidis isolate treated with proteinase-3 and another one without the enzyme were inserted into the back of a mouse. After 7 days of incubation period, the catheters were recovered and the number of grown bacteria was quantified, finding a higher amount of adhered proteinase-3-treated bacteria in the catheter than non-proteinase-3-treated bacteria (p < 0.05). Commensal non-biofilm-forming S. epidermidis in the presence of neutrophil cells significantly induced the biofilm formation when multiplicity of infection (MOI) 1:0.01 (neutrophil:bacteria) was used, but the addition of a cocktail of protease inhibitors impeded biofilm formation. A neutrophil:bacteria assay did not induce neutrophil extracellular traps (NETs). Our results suggest that neutrophils, in the presence of commensal non-biofilm-forming S. epidermidis, do not generate NETs formation. The effect of neutrophils is the production of proteases, and proteinase-3 releases bacterial DNA at the initial adhesion, favoring cell aggregation and subsequently leading to biofilm formation.     

The Effect of a Silver Nanoparticle Polysaccharide System on Streptococcal and Saliva-Derived Biofilms

In this work, we studied the antimicrobial properties of a nanocomposite system based on a lactose-substituted chitosan and silver nanoparticles: Chitlac-nAg. Twofold serial dilutions of the colloidal Chitlac-nAg solution were both tested on Streptococcus mitis, Streptococcus mutans, and Streptococcus oralis planktonic phase and biofilm growth mode as well as on saliva samples. The minimum inhibitory and bactericidal concentrations of Chitlac-nAg were evaluated together with its effect on sessile cell viability, as well as both on biofilm formation and on preformed biofilm. In respect to the planktonic bacteria, Chitlac-nAg showed an inhibitory/bactericidal effect against all streptococcal strains at 0.1% (v/v), except for S. mitis ATCC 6249 that was inhibited at one step less. On preformed biofilm, Chitlac-nAg at a value of 0.2%, was able to inhibit the bacterial growth on the supernatant phase as well as on the mature biofilm. For S. mitis ATCC 6249, the biofilm inhibitory concentration of Chitlac-nAg was 0.1%. At sub-inhibitory concentrations, the Streptococcal strains adhesion capability on a polystyrene surface showed a general reduction following a concentration-dependent-way; a similar effect was obtained for the metabolic biofilm activity. From these results, Chitlac-nAg seems to be a promising antibacterial and antibiofilm agent able to hinder plaque formation.     

One Step Forward with Dry Surface Biofilm (DSB) of Staphylococcus aureus: TMT-Based Quantitative Proteomic Analysis Reveals Proteomic Shifts between DSB and Hydrated Biofilm

The Gram-positive bacterium Staphylococcus aureus is responsible for serious acute and chronic infections worldwide and is well-known for its biofilm formation ability. Recent findings of biofilms on dry hospital surfaces emphasise the failures in current cleaning practices and disinfection and the difficulty in removing these dry surface biofilms (DSBs). Many aspects of the formation of complex DSB biology on environmental surfaces in healthcare settings remains limited. In the present study, we aimed to determine how the protein component varied between DSBs and traditional hydrated biofilm. To do this, biofilms were grown in tryptic soy broth (TSB) on removable polycarbonate coupons in the CDC biofilm reactor over 12 days. Hydrated biofilm (50% TSB for 48 h, the media was then changed every 48 h with 20% TSB, at 37 °C with 130 rpm). DSB biofilm was produced in 5% TSB for 48 h at 35 °C followed by extended periods of dehydration (48, 66, 42 and 66 h at room temperature) interspersed with 6 h of 5% TSB at 35 °C. Then, we constructed a comprehensive reference map of 12-day DSB and 12-day hydrated biofilm associated proteins of S. aureus using a high-throughput tandem mass tag (TMT)-based mass spectrometry. Further pathway analysis of significantly differentially expressed identified proteins revealed that proteins significantly upregulated in 12-day DSB include PTS glucose transporter subunit IIBC (PtaA), UDP-N-acetylmuramate-L-alanine ligase (MurC) and UDP-N-acetylenolpyruvoylglucosamine (MurB) compared to 12-day hydrated biofilm. These three proteins are all linked with peptidoglycan biosynthesis pathway and are responsible for cell-wall formation and thicker EPS matrix deposition. Increased cell-wall formation may contribute to the persistence of DSB on dry surfaces. In contrast, proteins associated with energy metabolisms such as phosphoribosyl transferase (PyrR), glucosamine--fructose-6-phosphate aminotransferase (GlmS), galactose-6-phosphate isomerase (LacA), and argininosuccinate synthase (ArgG) were significantly upregulated whereas ribosomal and ABC transporters were significantly downregulated in the 12-day hydrated biofilm compared to DSB. However, validation by qPCR analysis showed that the levels of gene expression identified were only partially in line with our TMT-MS quantitation analysis. For the first time, a TMT-based proteomics study with DSB has shed novel insights and provided a basis for the identification and study of significant pathways vital for biofilm biology in this reference microorganism.     

季铵盐改性蒙脱土的抗菌活性及抗菌机理

采用离子交换法将不同季钱盐插层到钠基蒙脱土中制备了改性蒙脱土.改性蒙脱土对革兰氏阳性菌(S.aereus)和革兰氏阴性菌(E coli)均有很强的抗菌作用,并且随着季铵盐在蒙脱土中的质量分数增加其抗菌活性增强.不同季铵盐改性蒙脱土的抗菌活性不同,其中双季铵盐改性蒙脱土的抗菌活性最好,对S.aereus和E coli的最小抑菌浓度分别为6.25 mg/L和12.5 mg/L.用扫描电镜对与细菌接触不同时间后的改性蒙脱土进行观察,结果表明:细菌先吸附到改性蒙脱土的表面,然后慢慢的变形死亡.同时,对在0.9%的生理盐水中浸泡不同时间后的改性蒙脱土进行X射线衍射和热重分析.结果表明:随着浸泡时间的增加,蒙脱土中的有机物质量分数及层间距均逐渐减少,说明季铵盐能从蒙脱土的层间解吸出来,并进入溶液中直接杀死细菌.因而,改性蒙脱土的抗菌活性是吸附与释放到溶液中的季铵盐离子协同作用的结果.     

Anti-Bacterial Effect of Cannabidiol against the Cariogenic Streptococcus mutans Bacterium: An In Vitro Study

Dental caries is caused by biofilm-forming acidogenic bacteria, especially Streptococcus mutans, and is still one of the most prevalent human bacterial diseases. The potential use of cannabidiol (CBD) in anti-bacterial therapies has recently emerged. Here we have studied the anti-bacterial and anti-biofilm activity of CBD against S. mutans. We measured minimum inhibitory concentration (MIC) and minimum biofilm inhibitory concentration (MBIC). The bacterial growth and changes in pH values were measured in a kinetic study. The biofilm biomass was assessed by Crystal Violet staining and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) metabolic assay. Spinning Disk Confocal Microscopy (SDCM) was used to assess biofilm structure, bacterial viability and extracellular polysaccharide (EPS) production. CBD inhibited S. mutans planktonic growth and biofilm formation in a dose-dependent manner, with similar MIC and MBIC values (5 µg/mL). CBD prevented the bacteria-mediated reduction in pH values that correlated with bacterial growth inhibition. SDCM showed a decrease of 50-fold in live bacteria and EPS production. CBD significantly reduced the viability of preformed biofilms at 7.5 µg/mL with an 80 ± 3.1% reduction of metabolic activity. At concentrations above 20 µg/mL, there was almost no bacterial recovery in the CBD-treated preformed biofilms even 48 h after drug withdrawal. Notably, precoating of the culture plate surfaces with CBD prior to incubation with bacteria inhibited biofilm development. Additionally, CBD was found to induce membrane hyperpolarization in S. mutans. Thus, CBD affects multiple processes in S. mutans including its cariogenic properties. In conclusion, we show that CBD has a strong inhibitory effect against cariogenic bacteria, suggesting that it is a potential drug adjuvant for reducing oral pathogenic bacterial load as well as protecting against dental caries.     

固化菌藻系统处理养殖废水中氨氮的研究

采用海藻酸钠-氯化钙固定法固化硝化细菌和硝化细菌与小球藻的混合物,来处理养殖废水中的氨氮污染,考察了处理时间、温度、p H和氨氮/固化小球用量比对氨氮去除效果的影响。实验结果显示,固化硝化细菌小球和固化菌藻小球均能有效去除废水中的氨氮,但固化菌藻小球的去除效果更佳。在28℃、p H=8、氨氮起始质量浓度为50 mg/L、氨氮/固化菌藻小球用量比为1∶40的实验条件下,24 h内能去除废水中96.51%的氨氮。实验结果证实,硝化细菌和小球藻具有一定的共生关系,在去除氨氮时有协同效应,固化菌藻小球在养殖废水脱氮中具有一定的应用前景。     

活性炭的改性条件及其对硫化氢吸附性能的影响

以工业活性炭为载体制备改性活性炭,对比研究了未改性活性炭,NaOH、Na2CO3、Fe(NO3)3、Cu(Ac)2改性活性炭及挂膜硫氧化细菌后活性炭在相同条件下对硫化氢穿透时间及吸附容量的影响。结果表明：在相同控制条件下,NaOH改性活性炭明显优于其它改性剂;不同梯度改性剂条件下,20% NaOH改性活性炭对硫化氢的吸附效果最好,吸附穿透容量为78.25 mg/g,穿透时间可以达到2000 min以上;不同改性剂挂膜硫氧化细菌后对硫化氢均有一定的处理效果,其中对已达到饱和吸附的NaOH改性活性炭挂膜后的再生效果可以达到100%以上,说明挂膜硫氧化细菌活性炭对硫化氢的处理具有很好的效果。     

1株高效聚磷菌的分离鉴定及固定化应用

为修复富营养化水体提供理论和实验依托,探讨了聚磷菌及固定化后对磷去除的影响,从氧化沟曝气池中筛选出1株高效聚磷菌W1,并经16SrDNA鉴定和系统发育分析,同时研究该菌株的除磷特性和固定化应用.结果表明,菌株W1确定为不动杆菌属(Acinetobacter sp.).W1生长较为迟缓,20 h进入对数期,40 h后进入...