Differential responses of cells from human skin keratinocyte and bovine mammary epithelium to attack by pore-forming Staphylococcus aureus alpha-toxin

Human skin keratinocytes HaCat attacked by Staphylococcus aureus α-toxin showed a transient drop of cellular ATP levels whereas in toxin-perforated bovine mammary epithelial cells (BMEC), the ATP levels dropped more slowly. Morphologically, during the ATP level depletion, HaCat cell developed a spacious intracellular vacuole together with the transient influx of trypan blue. WST-1 signal, which tested the function of mitochondrial enzyme in viable cells, also decreased concomitantly. On the other hand, BMEC excluded trypan blue and vacuolation was not observed throughout the experiment. We conclude that mammary epithelial cells resist the toxin better than keratinocytes. This is the first report showing that α-toxin enhances transient membrane permeability to large molecules, temporary vacuole formation and the transient defect of mitochondrial enzyme in viable cells without cell lysis.     

Genetic basis of penicillin resistance of S. aureus isolated in bovine mastitis

Background

The blaZ gene encoding penicillin resistance can be located either chromosomally or on plasmids. The aim of this study was to investigate the genetic relationships and to determine the location of the blaZ gene in S. aureus isolated in bovine mastitis in Finland and Sweden.

Methods

Seventy-eight β-lactamase positive S. aureus isolates from bovine mastitis (34 from Finland and 44 from Sweden) were included in the study. The localization of blaZ gene was determined by Southern blotting. The blaZ genes of the isolates were sequenced and the sequences were translated to beta-lactamase proteins and further grouped as different protein signatures. The isolates and, as control, 33 Swedish and 36 Finnish beta-lactamase negative isolates were typed with pulsed-field gel electrophoresis (PFGE).

Results

In 26 out of 34 Finnish isolates (76.5%) and in 25 out of 44 Swedish isolates (56.8%) the blaZ gene was localized on a plasmid. Six different protein signatures were found. One signature was found only in four Swedish isolates, but all other signatures were found both in Finnish and Swedish isolates. The PFGE results revealed a diversity of S. aureus clones. The protein signatures were not clearly associated with certain pulsotypes.

Conclusions

The plasmid location of the blaZ gene was not statistically significantly more common in Finland than in Sweden, and hence does not explain the higher proportion of penicillin-resistant isolates of S. aureus causing bovine mastitis in Finland compared to Sweden.     

猪胴体淋巴结中金黄色葡萄球菌的分离与鉴定

随机采取西宁市某生猪屠宰场猪胴体淋巴结60份,进行细菌常规分离培养,结果分离出金黄色葡萄球菌菌株40株,表明该批猪胴体中的金黄色葡萄球菌带菌率可能较高,应引起有关部门的重视。     

甘草总黄酮对金黄色葡萄球菌的作用研究

为了研究甘草总黄酮对金葡菌的抗菌活性以及亚抑菌浓度的甘草总黄酮对金黄色葡萄球菌α-溶血素分泌的影响。首先,采用微量肉汤稀释法测定了甘草总黄酮对6株金葡菌标准菌株的最小抑菌浓度;其次,采用溶血试验、蛋白免疫印迹试验以及荧光定量PCR试验分别在表型功能、蛋白表达和基因转录3个层次确证了其对α-溶血素表达的影响。结果表明：甘草总黄酮具有良好的抗金葡菌作用,其最小抑菌浓度范围为4~16 μg/mL,而亚抑菌浓度的甘草总黄酮能浓度依赖性地降低金葡菌α-溶血素的分泌。     

奥利亚罗非鱼在光场中的行为反应研究

为探索了解奥利亚罗非鱼在光场中的趋避行为规律。采用水平光梯度法,研究了4组不同生长阶段的奥利亚罗非鱼：Ⅰ组：20.0~50.0 mm;Ⅱ组：50.0~80.0 mm;Ⅲ组：80.0~110.0 mm;Ⅳ组：110.0~140.0 mm,在250、500、1000、1500、2000 lx白光光强梯度下的行为反应。同时对Ⅰ组奥利亚罗非鱼在不同波长单色光下（红、黄、绿、蓝）下的行为分布开展研究。结果表明：在白光下,4组奥利亚罗非鱼均表现出正趋光性,在250~2000 lx间趋光率随光照强度的增强呈现出先上升后下降的趋势,在1000 lx时趋光率达到峰值,其中近光区趋光率分别为67.5%、55.0%、55.0%、42.0%,总趋光率分别为96.0%、94.0%、96.5%、95.0%;奥利亚罗非鱼的白光适宜照度范围为：500~1500 lx;随着奥利亚罗非鱼的生长发育,近光区趋光率峰值Ⅰ>Ⅱ>Ⅲ>Ⅳ。此外,Ⅰ组奥利亚罗非鱼在不同单色光同一照度下的趋光率差异显著(P<0.05),在蓝光、绿光下的趋光率高于红光、黄光,对不同单色光的趋集行为具有选择性。研究表明：奥利亚罗非鱼在不同光色、不同光强、不同生长阶段都具有选择性,研究结果在开发高效诱捕灯、提高工厂化养殖效益等方面具有积极意义。     

乳制品中金黄色葡萄球菌PCR快速检测方法的建立

为快速检测乳制品中的金黄色葡萄球菌,本研究将纳米氢氧化钛和纳米氢氧化锆固定在滤纸片上以去除PCR反应抑制因子,并以Nuc为靶基因,建立一种无需样品前增菌的PCR快速检测方法,同时确定该方法的特异性、灵敏度和实用性,并评估该方法所用试剂在冷冻保存条件下的稳定性与实用性。结果表明,该PCR快速检测方法在检测97株目标菌及83株非目标菌后未出现假阳性或假阴性结果;该方法无需增菌处理,可在4 h内完成检测,检测限为10¹ CFU·25 g^-1或10¹ CFU·25 mL^-1;该方法与传统检测方法在实际乳制品样品中金黄色葡萄球菌的检出率及活菌检测率均一致(符合率100%);此外,该检测方法所用试剂在-20℃冷冻条件下可稳定保存至少12个月。综上所述,本研究建立的PCR快速检测方法特异性强、灵敏度高,且实用性良好,能有效去除样品基质中的PCR反应抑制因子,为乳制品中金黄色葡萄球菌的快速检测提供了参考依据。     

淡腌青鱼中木糖葡萄球菌碳源代谢能力研究

为探究淡腌青鱼的腐败与其微生物代谢能力的关系,利用Biolog GENⅢ微孔板对淡腌青鱼货架期终点的特定腐败菌(木糖葡萄球菌)在5、15、25、33℃4种温度下的碳源利用情况进行研究,采用修正的Gompertz模型对代谢曲线进行拟合,并以平均颜色变化率(AWCD)为指标研究其代谢速率,分析木糖葡萄球菌利用碳源能力的动力学特征。结果表明:木糖葡萄球菌能利用糖、氨基酸和羧酸类等碳源;相同温度下木糖葡萄球菌对碳源的代谢能力由大到小依次为糖、羧酸、氨基酸,其中单糖(葡萄糖、甘露糖)、双糖(蔗糖、海藻糖)、多糖及其糖类衍生物(N-乙酰-β-D-甘露糖胺、β-甲酰-D-葡糖)、氨基酸(谷氨酸、丝氨酸)和羧酸(L-乳酸)的代谢较强。木糖葡萄球菌在25℃下的延滞期最短,进入指数期最快。通过对木糖葡萄球菌在不同温度下各种碳源代谢能力的分析,为优化产品配方有效抑制微生物的活动、延长产品货架期提供理论依据。     

Activity of blue green microalgae extracts against in vitro generated Staphylococcus aureus with reduced susceptibility to vancomycin

Blue green microalgae have been identified as one of the promising groups of organism from which biochemically active natural products have been isolated. Aqueous and organic extracts of nine blue green microalgae strains were screened against in vitro generated vancomycin intermediate resistant Staphylococcus aureus (VISA) strains. Aqueous extracts of all the blue green microalgae cultures were found to be inactive, while all the organic (hexane, chloroform and methanolic) extracts of Anabaena virabilis and Anabaena sp. showed activity against VISA strains with MIC of 32-64 mug/ml.     

黑麦草-耳葡萄球菌对水体中Cu的联合修复效应研究

采用水培实验方法模拟不同程度的Cu污染状况,研究了黑麦草与耳葡萄球菌联合修复作用对Cu的去除效果。结果表明,联合修复作用对Cu的去除效果明显优于单一的黑麦草修复,Cu初始浓度在5-30mg.L-1时,联合修复对Cu的去除率均达85%以上。耳葡萄球菌对Cu表现出较强的耐受性和吸附性,Cu浓度为100mg.L-1时其干重积累量达到40.6mg.g-1。耳葡萄球菌的存在降低了黑麦草根系及茎叶中Cu的积累量,联合修复组的黑麦草的生物量和叶绿素含量明显高于单一黑麦草修复组。该菌在一定程度上减轻了黑麦草所受Cu的毒害性,提高了水中Cu的联合修复效果。     

缺硼对绿豆叶片光合特性和碳水化合物含量的影响

本试验以绿豆为指示植物,采用溶液培养法研究了缺硼对叶片光合功能和碳水化合物含量的影响,以明确缺硼是否影响光合作用进而影响作物生长。在绿豆第二片真叶出现后的第二天,一半植株供给 50 mol/L 硼（高硼）,另一半植株供给0.2 mol/L 硼（低硼）,在叶片生长过程中动态监测叶绿素含量、 气体交换和碳水化合物含量。结果表明,缺硼对叶片叶绿素含量没有影响,缺硼降低了光合速率（Pn）和气孔导度（Gs）,但对胞间二氧化碳浓度（Ci）没有影响; 缺硼提高了气孔限制率（Ls）。虽缺硼降低了Pn,但提高了叶片中可溶性糖,特别是葡萄糖和淀粉的含量。本研究结果表明缺硼对植物生长的影响并非是由于碳水化合物缺乏的缘故,而是因降低库活力导致了Pn的降低。