Efficiency of phenol biodegradation by planktonic Pseudomonas pseudoalcaligenes (a constructed wetland isolate) vs. root and gravel biofilm

In the last two decades, constructed wetland systems gained increasing interest in wastewater treatment and as such have been intensively studied around the world. While most of the studies showed excellent removal of various pollutants, the exact contribution, in kinetic terms, of its particular components (such as: root, gravel and water) combined with bacteria is almost nonexistent.In the present study, a phenol degrader bacterium identified as Pseudomonas pseudoalcaligenes was isolated from a constructed wetland, and used in an experimental set-up containing: plants and gravel. Phenol removal rate by planktonic and biofilm bacteria (on sterile Zea mays roots and gravel surfaces) was studied. Specific phenol removal rates revealed significant advantage of planktonic cells (1.04 × 10⁻⁹ mg phenol/CFU/h) compared to root and gravel biofilms: 4.59 × 10⁻¹¹-2.04 × 10⁻¹⁰ and 8.04 × 10⁻¹¹-4.39 × 10⁻¹⁰ (mg phenol/CFU/h), respectively.In batch cultures, phenol biodegradation kinetic parameters were determined by biomass growth rates and phenol removal as a function of time. Based on Haldane equation, kinetic constants such as μ_max = 1.15/h, K_s = 35.4 mg/L and K_i = 198.6 mg/L fit well phenol removal by P. pseudoalcaligenes.Although P. pseudoalcaligenes planktonic cells showed the highest phenol removal rate, in constructed wetland systems and especially in those with sub-surface flow, it is expected that surface associated microorganisms (biofilms) will provide a much higher contribution in phenol and other organics removal, due to greater bacterial biomass.Factors affecting the performance of planktonic vs. biofilm bacteria in sub-surface flow constructed wetlands are further discussed.     

Bacterial-based additives for the production of artificial snow: What are the risks to human health?

For around two decades, artificial snow has been used by numerous winter sports resorts to ensure good snow cover at low altitude areas or more generally, to lengthen the skiing season. Biological additives derived from certain bacteria are regularly used to make artificial snow. However, the use of these additives has raised doubts concerning the potential impact on human health and the environment. In this context, the French health authorities have requested the French Agency for Environmental and Occupational Health Safety (Afsset) to assess the health risks resulting from the use of such additives. The health risk assessment was based on a review of the scientific literature, supplemented by professional consultations and expertise. Biological or chemical hazards from additives derived from the ice nucleation active bacterium Pseudomonas syringae were characterised. Potential health hazards to humans were considered in terms of infectious, toxic and allergenic capacities with respect to human populations liable to be exposed and the means of possible exposure. Taking into account these data, a qualitative risk assessment was carried out, according to four exposure scenarios, involving the different populations exposed, and the conditions and routes of exposure. It was concluded that certain health risks can exist for specific categories of professional workers (mainly snowmakers during additive mixing and dilution tank cleaning steps, with risks estimated to be negligible to low if workers comply with safety precautions). P. syringae does not present any pathogenic capacity to humans and that the level of its endotoxins found in artificial snow do not represent a danger beyond that of exposure to P. syringae endotoxins naturally present in snow. However, the risk of possible allergy in some particularly sensitive individuals cannot be excluded. Another important conclusion of this study concerns use of poor microbiological water quality to make artificial snow.     

Evaluation of phosphate removal from water by immobilized phosphate-binding protein PstS

The phosphate (P(i))-binding protein PstS is a member of a family of periplasmic proteins that act as high-affinity receptors for active transport systems in bacteria. PstS protein purified from Pseudomonas aeruginosa was immobilized to N-hydroxysuccinimide-activated Sepharose, packed into a plastic column (5 x 70 mm), and examined for its potential ability to remove P(i) from water. The PstS-Sepharose column completely removed P(i) from 32P-labeled pond water containing about 0.5 microM P(i) (0.015 mg P per liter). More than 90% of 32P-P(i) that was retained in the column could be eluted by washing with low-pH water.     

烟草青枯病菌(Pseudomonas solanacearum)细菌素的生物学特性研究

本文研究了烟草青枯病菌(Pseudomonas solanacearum)无毒产细菌素菌株 A_(3-5)产生的细菌素的生物学特性,结果表明,用不同制备方法提取的 A_(3-5)的细菌素,Au 值均为8。失活温度为60℃,在30℃以下温度放置25天抑菌活性不变,35℃下放置20天,活性开始减弱,40℃放置10天,活性即减弱,25天对几乎完全失活。这说明 A_(3-5)的细菌素虽具有较强耐热能力,但高温不利于其存活。此外,0.5%的胰蛋白酶即刻使其完全失活,说明对胰蛋白酶敏感。经1万转/分钟和5万转/分钟连续离心后,细菌素仍存在于上清液中,说明为非沉淀性的。以上特性表明 A_(3-5)的细菌素可能是一种小分子物质。     

Purification, characterization, and steady-state kinetics of a meta-cleavage compound hydrolase from Pseudomonas fluorescens IPO1

2-Hydroxy-6-oxo-7-methylocta-2,4-dienoate (6-isopropyl-HODA) hydrolase (CumD), an enzyme of the cumene biodegradation pathway encoded by the cumD gene of Pseudomonas fluorescens IP01, was purified to homogeneity from an overexpressing Escherichia coli strain. SDS-polyacrylamide gel electrophoresis and gel filtration demonstrated that it is a dimeric enzyme with a subunit molecular mass of 32 kDa. The pH optima for activity and stability were 8.0 and 7.0-9.0, respectively. The enzyme exhibited a biphasic Arrhenius plot of catalysis with two characteristic energies of activation with a break point at 20 degrees C. The enzyme has a K(m) of 7.3 microM and a k(cat) of 21 s(-1) for 6-isopropyl-HODA (150 mM phosphate, pH 7.5, 25 degrees C), and its substrate specificity covers larger C6 substituents compared with another monoalkylbenzene hydrolase, TodR Unlike TodF, CumD could slightly hydrolyze 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate (6-phenyl-HODA). A mutant enzyme as to a putative active site residue, S103A, had 10(5)-fold lower activity than that of the wild-type enzyme.     

荧光假单胞菌和热杀索丝菌对低温贮藏期间猪肉品质变化的影响

为研究荧光假单胞菌和热杀索丝菌与猪肉品质变化的相关性,测定了在4 ℃下贮藏猪肉的微生物菌落数、pH值、色泽（L*、a*、b*值）、质构特性、总糖含量、总挥发性盐基氮（total volatile basic nitrogen,TVB-N）值和硫代巴比妥酸反应物（thiobarbituric acid reactive substances,TBARS）值以及通过扫描电子显微镜观察猪肉肌纤维微观结构的变化。结果表明,在冷藏期间,微生物菌落数、pH值、TVB-N值、TBARS值等随着猪肉冷藏时间延长而上升,而总糖含量、L*值、a*值、硬度、咀嚼性均呈降低趋势。同时,研究发现猪肉冷藏过程中微生物的生长繁殖使其肌纤维结构发生明显变化。猪肉理化品质变化与微生物的种类和生长速率相关,热杀索丝菌的致腐败能力高于假单胞菌。相关性分析结果表明,菌落数、pH值、总糖含量、TVB-N值、TBARS值与冷藏时间密切相关,其中微生物菌落数与冷藏时间的相关性最高,可作为冷藏猪肉品质评价和货架期预测的监测指标。     

不同细菌胞外产物对腐败希瓦氏菌低温下生长代谢的影响

为研究荧光假单胞菌(Pseudomonas fluorescens,P.fluorescens)、嗜水气单胞菌(Aeromonas hydrophila,A.hydrophila)、温和气单胞菌(Aeromonas sobria,A.sobria)的胞外产物对腐败希瓦氏菌(Shewanella putrefaciens,S.putrefaciens)低温下生长的影响,提取这3株菌株的胞外产物与无菌脑心浸肉汤(Brain Heart Infusion,BHI)培养基混合,再将S.putrefaciens分别接种至混合培养液中,于(4±1)℃下培养12d,测定培养液中S.putrefaciens的菌落总数及胞外蛋白酶、生物膜、挥发性盐基氮(TVB-N)、总氨基酸和腐胺的变化。结果表明:P.fluorescens、A.hydrophila和A.sobria的胞外产物对S.putrefaciens的延滞期和对数期几乎没有影响,但是A.sobria的胞外产物可以抑制S.putrefaciens稳定期的生长,而且随着贮藏时间的延长,相比于P.fluorescens、A.hydrophila,A.sobria的胞外产物对S.putrefaciens的生物膜、胞外蛋白酶形成以及致腐败能力的抑制作用更强。     

Detection and quantification of Escherichia coli and Pseudomonas aeruginosa in cow milk by near‐infrared spectroscopy

This work investigated the potential of NIR technology to be applied in the dairy industry for the detection of micro‐organisms. To this end, two types of cow milk samples were studied, one in which only bacterial biomass was considered and the other in which bacteria were cultured and grown in milk for 24 h. The study was carried out using two micro‐organisms Escherichia coli and Pseudomonas aeruginosa. Both types of samples with different counts of both micro‐organisms were analysed by a NIR analyser in the range 10 000–4000 cm^?1 based on transmittance measurements. Multivariate models indicated that a better discrimination between micro‐organisms was attained in those milk samples in which micro‐organisms have been grown.     

Synthesis of Mixed Refrigerant Cascade Cycles

This paper considers the synthesis of refrigeration systems with refrigerant mixtures as working fluids. The employed refrigeration topology encompasses features from industrial liquefaction of natural gas (LNG) systems. This configuration consists of a combination of horizontal and vertical cascades generalizing the vertical cascade of pure refrigerant systems. The key features of mixtures exploited here are their ability to evaporate/condense over a temperature range and their potential to generate streams with different compositions through partial condensation. The synthesis problem is formulated and solved as a nonlinear program. The proposed methodology is illustrated using an example of cooling a methane-rich stream.