Coagulase-negative staphylococci species affect biofilm formation of other coagulase-negative and coagulase-positive staphylococci

Coagulase-negative staphylococci (CNS) are considered to be commensal bacteria in humans and animals, but are now also recognized as etiological agents in several infections, including bovine mastitis. Biofilm formation appears to be an important factor in CNS pathogenicity. Furthermore, some researchers have proposed that CNS colonization of the intramammary environment has a protective effect against other pathogens. The mechanisms behind the protective effect of CNS have yet to be characterized. The aim of this study was to evaluate the effect of CNS isolates with a weak-biofilm phenotype on the biofilm formation of other staphylococcal isolates. We selected 10 CNS with a weak-biofilm phenotype and 30 staphylococcal isolates with a strong-biofilm phenotype for this study. We measured biofilm production by individual isolates using a standard polystyrene microtiter plate assay and compared the findings with biofilm produced in mixed cultures. We confirmed the results using confocal microscopy and a microfluidic system with low shear force. Four of the CNS isolates with a weak-biofilm phenotype (Staphylococcus chromogenes C and E and Staphylococcus simulans F and H) significantly reduced biofilm formation in approximately 80% of the staphylococcal species tested, including coagulase-positive Staphylococcus aureus. The 4 Staph. chromogenes and Staph. simulans isolates were also able to disperse pre-established biofilms, but to a lesser extent. We also performed a deferred antagonism assay and recorded the number of colony-forming units in the mixed-biofilm assays on differential or selective agar plates. Overall, CNS with a weak-biofilm phenotype did not inhibit the growth of isolates with a strong-biofilm phenotype. These results suggest that some CNS isolates can negatively affect the ability of other staphylococcal isolates and species to form biofilms via a mechanism that does not involve growth inhibition.     

The Potential Source of B. licheniformis Contamination During Whey Protein Concentrate 80 Manufacture

The objective of this study was to determine the possible source of predominant Bacillus licheniformis contamination in a whey protein concentrate (WPC) 80 manufacturing plant. Traditionally, microbial contaminants of WPC were believed to grow on the membrane surfaces of the ultrafiltration plant as this represents the largest surface area in the plant. Changes from hot to cold ultrafiltration have reduced the growth potential for bacteria on the membrane surfaces. Our recent studies of WPCs have shown the predominant microflora B. licheniformis would not grow in the membrane plant because of the low temperature (10 °C) and must be growing elsewhere. Contamination of dairy products is mostly due to bacteria being released from biofilm in the processing plant rather from the farm itself. Three different reconstituted WPC media at 1%, 5%, and 20% were used for biofilm growth and our results showed that B. licheniformis formed the best biofilm at 1% (low solids). Further investigations were done using 3 different media; tryptic soy broth, 1% reconstituted WPC80, and 1% reconstituted WPC80 enriched with lactose and minerals to examine biofilm growth of B. licheniformis on stainless steel. Thirty‐three B. licheniformis isolates varied in their ability to form biofilm on stainless steel with stronger biofilm in the presence of minerals. The source of biofilms of thermo‐resistant bacteria such as B. licheniformis is believed to be before the ultrafiltration zone represented by the 1% WPC with lactose and minerals where the whey protein concentration is about 0.6%.     

鳀鱼抗菌肽脂质体的制备与抗单增李斯特菌生物被膜活性研究

本文采用薄膜蒸发法制备得到鳀鱼抗菌肽AAP脂质体,分析了脂质体的平均粒径、形态结构、包封率和贮存稳定性,研究了其对单增李斯特菌及其生物被膜的抑制活性。结果表明制备得到的AAP脂质体平均粒径为(131.65±1.63)nm,包封率为69.75%,AAP有效载量为4.17%,为内部呈环形层状分布的近球形。脂质体在4℃下的贮存稳定性高于其在25℃下的稳定性。脂质体型AAP和未包封AAP均能抑制单增李斯特菌的生长,但因脂质体的控释作用,两者对指示菌生长曲线的改变历程略有差异。脂质体型AAP抗单增李斯特菌生物被膜活性高于未包封AAP。结晶紫染色和银染实验结果表明AAP脂质体能抑制单增李斯特菌生物被膜的形成。扫描电镜和激光共聚焦显微镜形态观察表明AAP脂质体可引起细菌细胞结构的明显坍塌、细胞膜破损和胞内物质外溢,从而抑制生物被膜的形成。     

氨曲南联用阿奇霉素对产膜铜绿假单胞菌作用的实验研究

目的:探讨氨曲南联用阿奇霉素对铜绿假单胞菌ATCC 27853及临床分离的15株铜绿假单胞菌的体外作用效果。方法: 分别采用肉汤稀释法、棋盘法测定氨曲南、阿奇霉素单用及两者联用对铜绿假单胞菌ATCC 27853、临床分离的15株铜绿假单胞菌的作用效果。刚果红平板实验鉴定临床分离的15株铜绿假单胞菌能否形成生物膜。结晶紫实验比较铜绿假单胞菌形成生物膜能力的强弱。连续稀释法绘制铜绿假单胞菌生物膜生长曲线及活菌计数。银染法观察氨曲南联用阿奇霉素对铜绿假单胞菌生物膜的情况。结果: 体外实验结果表明,氨曲南联用阿奇霉素,对铜绿假单胞菌ATCC 27853表现为协同作用;在临床分离的15株铜绿假单胞菌中,有6株表现为协同作用、有6株表现为相加作用、有3株表现为无关作用。刚果红平板实验表明,临床分离的15株铜绿假单胞菌中,有12株形成生物膜,3株未形成生物膜,成膜率为80%。结晶紫实验表明,编号16091217的菌株,与其他菌株相比,成膜能力最强。连续稀释法行活菌计数实验表明,阿奇霉素组、氨曲南组活菌数量与阿奇霉素+氨曲南组相比,有统计学差异。结论:氨曲南联用阿奇霉素对铜绿假单胞菌的体外作用以协同和相加作用为主,对能形成生物膜的铜绿假单胞菌,有协同杀菌作用。     

碳材料在抗海洋生物污损领域中应用的研究进展

海洋生物污损一直是困扰全球海洋经济发展的重要问题之一。任何材料进入海洋环境,其表面不可避免地会遭受蛋白、多糖、细菌、海藻、软体动物等海洋生物贴附形成生物污损,不仅影响海上装备的正常运转和增加能源消耗,同时极易引起微生物腐蚀。因此,海洋抗污损新材料的开发显得极为迫切。目前碳纳米材料因具有优异的理化特性,而在海洋防腐、耐磨等工程上大量使用,其抑制海洋污损物贴附和生长的性能决定了碳材料在抗海洋生物污损领域具有广阔的应用前景。综述了石墨烯、类金刚石、金刚石、碳纳米管、富勒烯及其衍生物等典型碳纳米材料,在抗蛋白质、细菌、海藻等生物污损领域的应用现状以及抗污损机理。碳材料在海洋环境中优异的综合性能,预示其将给抗海洋生物污损新材料的开发和应用带来更多的机遇和可能。     

4种防腐剂对副溶血弧菌生物膜形成的抑制作用

在确定4种防腐剂壳聚糖、山梨酸钾、脱氢乙酸钠和ε-聚赖氨酸对副溶血弧菌(Vibro parahaemolyticus)的最小抑菌浓度(minimum inhibitory concentration,MIC)的基础上,研究其对副溶血弧菌生物膜的抑制作用。采用结晶紫染色法测生物膜形成量,XTT法测生物膜代谢活性,硫酸-苯酚法测定生物膜中胞外多糖的分泌量。结果表明,壳聚糖的MIC最小,为1.25 mg/m L。4种防腐剂在MIC以及亚抑菌浓度条件下对副溶血弧菌生物膜均有明显的抑制作用,不仅抑制生物膜的形成,而且能显著降低细菌的代谢活性,减少胞外多糖的分泌,其中壳聚糖对副溶血弧菌生物膜的抑制作用最强。     

不同培养条件及群体感应信号分子对蜂房哈夫尼亚菌生物被膜的影响

为研究蜂房哈夫尼亚菌(Hafnia alvei)Ha-01生物被膜形成的过程及不同培养条件(碳源、p H值、Na Cl质量分数、黏附材料)对其生物被膜形成的影响,采用超声波平板法及扫描电镜法研究不同培养条件下菌株Ha-01生物被膜活菌数及生长情况,并通过添加外源标准群体感应信号分子(N-酰基高丝氨酸内酯(N-acyl-homoserine lactones,AHLs))中的C6-HSL研究了AHLs与其生物被膜形成的关系。结果显示,菌株Ha-01生物被膜的形成与培养时间密切相关;在不同碳源的培养条件下形成生物被膜能力不同,其中在以木糖为培养基时形成量最大,达到7.51(lg(CFU/cm~2)),与在LB培养基中相比增加10.28%;在中性培养条件下更利于其生物被膜的形成,活菌数为7.77(lg(CFU/cm~2));在Na Cl质量分数为2%时,其生物被膜产生量最大,活菌数为7.18(lg(CFU/cm~2));在不同黏附材料上生物被膜形成能力从大到小依次为铝片、锌片、玻璃片,活菌数分别为7.22、6.48、6.11(lg(CFU/cm~2));且添加C6-HSL量越多,其生物被膜产生能力越强。研究表明,培养条件能够影响菌株Ha-01生物被膜形成,且AHLs可以调控其生物被膜形成。     

培养条件对金黄色葡萄球菌-大肠杆菌混合生物被膜的影响

以常见食源性致病菌金黄色葡萄球菌和大肠杆菌为研究对象,采用微孔板法对混合生物被膜形成过程中的不同影响因素进行了研究。结果表明：在25 ℃或30 ℃条件下,采用质量分数0.8%的胰酶大豆肉汤（TSB）或0.8%的脑心浸出液（BHI）培养基培养时,生物被膜形成量较高;添加适量的葡萄糖、乳糖、麦芽糖和蔗糖可提高混合生物被膜形成能力;培养基中NaCl质量分数＞8%时,混合生物被膜的生长明显受到抑制。