Phytoplankton primary production, chlorophyll-a and nutrient concentrations in the water column of mountainous Lake Phewa, Nepal

The seasonal patterns of phytoplankton primary production, chlorophyll‐a concentration, cell number and several other limnological variables in Lake Phewa, located in the active monsoon zone in Central Himalaya, Nepal, were studied for a year beginning in April 2001. During the study period, the gross primary production and chlorophyll‐a concentrations were relatively low during the monsoon season. The phytoplankton cell number, represented by 24 genera, also fluctuated seasonally, but tended to increase in the pre‐ and post‐monsoon period. These results suggest that the monsoon plays a crucial role in the primary production and phytoplankton dynamics for Lake Phewa. Among the phytoplankton species, Microcystis aeruginosa, a representative species for eutrophic lakes, was the dominant phytoplankton. At the same time, however, it is clear that the lake is not yet heavily eutrophic. The present study suggests that the exchange of lake water during the monsoon season contributes to maintaining the health of the lake against further degradation. Nevertheless, the silt carried in the monsoon rain run‐off from the lake's catchment area suggests increasingly serious degradation problems for this small mountainous lake.     

Microstructured Lipid Carriers (MLC) Based on N-Acetylcysteine and Chitosan Preventing Pseudomonas aeruginosa Biofilm

The aim of this work was the development of microstructured lipid carriers (MLC) based on chitosan (CH) and containing N-acetylcysteine (NAC), a mucolytic and antioxidant agent, to inhibit the formation of Pseudomonas aeruginosa biofilm. MLC were prepared using the high shear homogenization technique. The MLC were characterized for morphology, particle size, Z potential, encapsulation efficiency and drug release. The antioxidant properties of NAC-loaded microstructured carriers were evaluated through an in vitro spectrophotometer assay. Finally, the activity of NAC-CH-MLC on biofilm production by Pseudomonas aeruginosa was also evaluated. Results obtained from this study highlighted that the use of chitosan into the inner aqueous phase permitted to obtain microstructured particles with a narrow size range and with good encapsulation efficiency. NAC-loaded MLC showed higher antioxidant activity than the free molecule, demonstrating how encapsulation increases the antioxidant effect of the molecule. Furthermore, the reduction of biofilm growth resulted extremely high with MLC being 64.74% ± 6.2% and 83.74% ± 9.95%, respectively, at 0.5 mg/mL and 2 mg/mL. In conclusion, this work represents a favorable technological strategy against diseases in which bacterial biofilm is relevant, such as cystic fibrosis.     

The Esterase PfeE,the Achilles’ Heel in the Battle for Iron between Pseudomonas aeruginosa and Escherichia coli

Bacteria access iron, a key nutrient, by producing siderophores or using siderophores produced by other microorganisms. The pathogen Pseudomonas aeruginosa produces two siderophores but is also able to pirate enterobactin (ENT), the siderophore produced by Escherichia coli. ENT-Fe complexes are imported across the outer membrane of P. aeruginosa by the two outer membrane transporters PfeA and PirA. Iron is released from ENT in the P. aeruginosa periplasm by hydrolysis of ENT by the esterase PfeE. We show here that pfeE gene deletion renders P. aeruginosa unable to grow in the presence of ENT because it is unable to access iron via this siderophore. Two-species co-cultures under iron-restricted conditions show that P. aeruginosa strongly represses the growth of E. coli as long it is able to produce its own siderophores. Both strains are present in similar proportions in the culture as long as the siderophore-deficient P. aeruginosa strain is able to use ENT produced by E. coli to access iron. If pfeE is deleted, E. coli has the upper hand in the culture and P. aeruginosa growth is repressed. Overall, these data show that PfeE is the Achilles’ heel of P. aeruginosa in communities with bacteria producing ENT.     

重组铜绿假单胞菌外毒素A工程菌发酵及表达产物的纯化

目的建立稳定、高效表达重组铜绿假单胞菌外毒素A(rEPA)的工程菌发酵及表达产物纯化工艺。方法规模化发酵表达rEPA的重组大肠杆菌rPE553D,离心收集菌体,渗透压调解使菌体周质间隙蛋白释放后,高速离心收集蛋白溶液。经DEAESepharoseFF、PhenylSepharose6FF疏水层析和SOURCE30Q强离子交换层析,超滤浓缩纯化rEPA。用HPLC、SDS-PAGE和Westernblot等方法检定生化和免疫学特性,并用小鼠和Vero细胞检定毒性。结果每55L培养基中rEPA产量超过4g,纯度在95%以上,细胞毒性降低了至少32000倍。其余各项指标均符合《中国生物制品规程》要求。结论已建立了收率高、纯度好、稳定、适合规模化生产rEPA的工艺。     

绿脓杆菌外膜蛋白对感染的保护作用

目的 寻找一种防治绿脓杆菌感染的有效方法。方法 用绿脓杆菌外膜蛋白免疫的兔抗血清对4种不同血清型的菌株和1株临床绿脓杆菌进行体内外交叉保护性试验。结果 家兔免疫1周后即产生抗体,并逐渐升高,至第6周抗体滴度达到1:163 840,并维持高水平;抗血清能与4种不同血清型菌株和1株临床菌株产生直接凝集反应。第8周的血清(1:8以上)与其外胰蛋白的ELISA抗体效价相近,小鼠的半保护剂量均在0.05～0.11ml之间。结论 外膜蛋白疫苗具有较强的抗原性,而且具有良好的交叉保护作用。     

高效产表面活性剂菌株（Lz2～1）的筛选及其特性研究

从24份含油土壤和水等样品中,经富集培养、摇瓶培养和排油活性测定等方法筛选出一株能以原油为碳源产生表面活性剂的菌株Lz2—1;该菌株可以将水的表面张力由72mN／m降到28mN／m,且发酵液具有较好的乳化活性;经生理生化16SrDNA及生理生化实验确定该菌株为铜绿假单胞菌;提取其代谢产物经薄层层析及红外色谱分析显示,主要的表面活性剂类物质为鼠李糖脂类,其临界胶束质量浓度为0．63047g／L。结果表明,表面活性物质是Lz2—1菌株在微生物采油过程中发挥作用的主要因素。     

掺硼金刚石薄膜电极电化学氧化对铜绿微囊藻的生长抑制

阳极材料采用掺硼金刚石薄膜板状电极,研究了电化学氧化中电流密度、电解时间、pH、氯离子浓度、硫酸根离子浓度对铜绿微囊藻生长抑制的影响,以及电解前后藻细胞形态的变化。结果表明,4个影响因素对铜绿微囊藻生长抑制效果显著。抑藻效果随电流密度、电解时间的增加而增加,电流密度为17 mA/cm²时藻细胞出现破裂、细胞内物质流出的现象,抑藻效果较好;当电解时间为20 min时,可完全抑制藻细胞生长;再增大电解时间,对抑藻效果无明显促进作用,初始pH在中性及酸性条件下可完全抑制藻细胞生长。抑藻效果与溶液中氯离子、硫酸根离子浓度成正相关,当溶液中氯离子浓度为6 mg/L时,可完全抑制藻细胞生长;无氯离子时,藻细胞在4 d后出现继续增长现象。     

一株铜绿微囊藻溶藻菌的溶藻机理研究

以铜绿微囊藻为溶解对象,从巢湖中分离到1株溶藻菌H2。探讨了菌株H2对铜绿微囊藻的溶藻方式,菌株H2胞外活性物质的溶藻特性。结果表明,菌株H2是间接溶藻。溶藻菌起溶藻作用的胞外分泌液中活性物质主要为分子量小于3.5k D的物质,并且活性物质是混合物质;菌株胞外分泌液经乙醇沉淀后,上清液部分的溶藻率大于沉淀部分,为52.50%;胞外分泌液经不同温度和酸碱度处理后,在25℃、p H=12.0时,活性较强,溶藻率分别为98.0%和97.1%;胞外分泌液中活性物质具有一定的抗氧化性。     

不同预氧化剂对铜绿微囊藻细胞的灭活

为比较不同预氧化剂对藻细胞的处理效果及其机理,在高锰酸钾灭活藻细胞研究的基础上,采用次氯酸钠与臭氧对铜绿微囊藻进行灭活实验.结果表明:次氯酸钠和臭氧对藻类的灭活反应均符合二级动力学模型;次氯酸钠较易引起铜绿微囊藻细胞萎缩破裂,从而导致细胞内代谢有机质的释放,对藻细胞的灭活动力学常数为(220±3)L·mol-1·s-1,不同初始次氯酸钠浓度对动力学常数无明显影响;臭氧对于铜绿微囊藻细胞有极强的氧化作用,细胞灭活速率常数达(2 655±15)L·mol-1·s-1,能迅速引起藻细胞破裂及胞内代谢有机质的释放,不同初始臭氧浓度对动力学常数无明显影响.3种氧化剂对铜绿微囊藻细胞的灭活速率由大到小依次为:臭氧次氯酸钠高锰酸钾.电镜扫描结果显示,臭氧与次氯酸钠在氧化过程中比高锰酸钾更易引起藻细胞破裂及IOM的释放.     

NY3菌固定化及生物膜处理含油废水的研究

利用嗜油菌NY3菌的固定化生物膜处理含油废水.用静态曝气法确定固定化时间、载体量、固定化液初始pH值.确定生物膜处理运行的HRT,并评估生物膜循环使用的可能性.结果表明,载体10 g/L,pH7.5~9.0,固定51 h,膜生物量达488.32 mg/g.其处理含油废水最佳HRT为6 h,出水油量在1.38~3.2 mg/L.废水处理后,将载体置于营养液中培养15 d,膜上NY3菌能将所吸附的油降解,使其恢复活性,并可循环利用.