Polysaccharide differences between planktonic and biofilm-associated EPS from Pseudomonas fluorescens B52

The polysaccharides associated with free (planktonic) and surface-attached (biofilm) cells from cultures of Pseudomonas fluorescens strain B52 were compared. Variations in the attached matrix due to surface material (glass or stainless steel) were also analyzed. Two digestion methods were used to optimize the recoveries of sugars, uronic acids and acidic substituents. The yield of analyzable material after digestion reached 90% for the material associated to the biofilms, though only 20–30% for that bound to planktonic cells. The polysaccharide(s) in the biofilm had glucuronic and guluronic acids as main components, besides rhamnose, glucose and glucosamine. The proportion of glucuronic to guluronic acid was higher in the polysaccharide(s) found in biofilms formed on stainless steel than in those on glass.     

Evaluation of epifluorescence image analysis of biofilm growth on stainless steel surfaces

Biofilm growth of Bacillus subtilis, Pseudomonas fragi, Pediococcus inopinatus and Listeria monocytogenes was studied on stainless steel surfaces at room and low temperatures to evaluate the results of traditional hygiene measures. The results were compared with those of image analysis of stainless steel surfaces in an epifluorescence microscope. Statistical analyses were carried out to determine the variations between the conventional cultivation swab method, the glycocalyx amount obtained using swabbing, and the values of the areas of the biofilm, slime and cells. As a general rule, old biofilms showed total counts at approximately the same levels as the young biofilm. The results showed that temperature affected the results for all strains except B. subtilis. The strains of Pe. inopinatus and Ps. fragi showed increased attachment at 6°C and L. monocytogenes at 25°C. The biofilm slime was more easily detached than the cells. The results indicated that the traditional swab method is not reliable for the measurement of biofilm formation on surfaces.     

Self-assembly and morphological characterization of two-component functional amyloid proteins

Functional amyloid has been increasingly applied as self-assembling nanostructures to construct multifunctional biomaterials. However, little has been known how different side domains, varied fusion positions and subunits affect self-assembly and morphologies of amyloid fibrils. Here, we constructed three groups of two-component amyloid proteins based on CsgA, the major protein components of Escherichia coli biofilms, to bridge these gaps. We showed that all fusion proteins have amyloid features, as indicated by Congo red assay. Atomic force microscopy (AFM) indeed reveals that these fusion proteins are able to self-assemble into fibrils, with an average diameter of 0.5-2 nm and length of hundreds of nanometers to several micrometers. The diameter of fibrils increases with the increase of the molecular weight of fusion domains, while the dynamic assembly of recombinant proteins was delayed as a result of the introduction of fusion domains. Moreover, fusion of the same functional domains but at intermediate position seems to cause the most interference on fibril assembly compared with those fused at C or Nterminus, as mainly short and irregular fibrils were detected. This phenomenon appears more pronounced for randomly coiled mussel foot proteins (Mfps) than for rigid chitin-binding domain (CBD). Finally, increase of the molecular weight of tandem repeats in protein monomer seemed to increase the fibril diameter of the resultant fibrils, but either reduction of the tandem repeats of CsgA to one single belta-sheet loop or increase in the number of tandem repeats of CsgAs from one to four produced shorter and intermittent fibrils compared with CsgA control protein. These studies therefore provide insights into self-assembly of two-component amyloid proteins and lay the foundation for rational design of multifunctional molecular biomaterials.     

Predation, death, and survival in a biofilm: Bdellovibrio investigated by atomic force microscopy

Biofilms are complex microbial communities that are resistant to attack by bacteriophages and to removal by drugs and chemicals. Here we use atomic force microscopy (AFM) to image the attack on Escherichia coli biofilms by Bdellovibrio bacteriovorus 109J. Bdellovibrio is a small, predatory bacterium that invades and devours other Gram-negative bacteria. We demonstrate that under dilute nutrient conditions, bdellovibrios can prevent the formation of simple bacterial biofilms and destroy established biofilms; under richer conditions the prey bacteria persist and are not eradicated, but may be shifted toward solution populations. Using AFM we explore these bacterial interactions with more detail and accuracy than available by more traditional staining assays or optical microscopy. AFM also allows us to investigate the nanoscale morphological changes of the predator, especially those related to motility. This demonstration of Bdellovibrio's successful predation in a biofilm inspires us to consider ways that it might be used productively for industrial, medical, agricultural, and biodefensive purposes.     

不同密度人工水草对氮、磷去除效果研究

污水处理厂出水作为城市公园景观用水,已成为我国北方缺水城市的通遍做法。由于污水处理厂出水中氮、磷含量相对较高,景观水体极易发生富营养化。本实验利用碳纤维人工水草作为生态基形成生物膜,对景观水体进行脱氮除磷处理效果进行研究。试验共布设2株/m~2、4株/m~2、10株/m~2、16株/m~2、20株/m~2 5种水草密度,通过SPSS进行方差分析,结果显示不同密度人工水草对氮,磷去除效果存在差异,明4株/m~2、10株/m~2,20株/m~2对总氮、氨氮、溶解性磷酸盐具有显著去除效果,对总磷去除效果不明显。叶绿素a浓度与氮、磷去除率呈正相关,表明在流动水体中藻类优势群落的形成对微生物膜快速形成以及污染物的去除有重要影响。     

Preparation of azidoaryl- and azidoalkyloxazoles for click chemistry

A series of azidoaryl- and azidoalkyl(diphenyl)oxazole scaffolds were warranted for biofilm inhibition studies. Cyclization of azidoaryl- or azidoalkyl esters of benzoin with ammonium acetate in acetic acid gives 2-azidoaryl- or 2-azidoalkyl-4,5-diphenyloxazoles. The azidoaryl esters are prepared from the corresponding azidocarboxylic acids/acid chlorides while the azidoalkyl esters are prepared from the corresponding haloalkyl esters.     

Polysaccharide differences between planktonic and biofilm-associated EPS from Pseudomonas fluorescens B52

The polysaccharides associated with free (planktonic) and surface-attached (biofilm) cells from cultures of Pseudomonas fluorescens strain B52 were compared. Variations in the attached matrix due to surface material (glass or stainless steel) were also analyzed. Two digestion methods were used to optimize the recoveries of sugars, uronic acids and acidic substituents. The yield of analyzable material after digestion reached 90% for the material associated to the biofilms, though only 20–30% for that bound to planktonic cells. The polysaccharide(s) in the biofilm had glucuronic and guluronic acids as main components, besides rhamnose, glucose and glucosamine. The proportion of glucuronic to guluronic acid was higher in the polysaccharide(s) found in biofilms formed on stainless steel than in those on glass.     

Detection of algae and bacterial biofilms formed on titanium surfaces using micro-Raman analysis

Biofouling is one of the major impediments in the use of titanium in sea-water cooled condensers of power plants, which is otherwise an excellent material with respect to corrosion resistance. Raman microscopic experiments were carried out on biofilms grown on titanium surfaces to find out the chemical composition of complex extracellular polymeric substances (EPS) in the biofilm. Though the spectral resolution of normal Raman experiments on these systems was very poor, it was improved when micro-SERS experiments were carried out using mono and bimetallic Ag and Cu colloids. It was observed that spatial distribution of polysaccharides was higher than that of proteins in algae biofilms formed on titanium matrix. Similar experiments were performed on laboratory cultured bacterial films of Pseudomonas aeruginosa. It was evidenced that algal and bacterial biofilms on titanium can be clearly distinguished with the help of Raman mapping coupled with SERS technique using bimetallic Ag/Cu colloids.     

Holographic microrheology of polysaccharides from <Emphasis Type="Italic">Streptococcus mutans</Emphasis> biofilms

We use three-dimensional holographic particle tracking to perform microrheological measurements of model gelled media, including the polysaccharide pellicle of dental biofilms created by the common cariogenic oral pathogen Streptococcus mutans. Nanometer-resolution video-rate holographic tracking of embedded colloidal spheres provides accurate measurements of the gels’ complex viscoelastic moduli, including insights into these properties’ heterogeneity. When applied to polysaccharides of S. mutans biofilms, these techniques promise quantitative microscopic assays for candidate therapeutic agents against cariogenic dental biofilms.     

铅和镉在实验室水环境模拟体系中的分布特征研究

选取长春市南湖作为采样地点,通过建立实验室水环境模拟体系(包括人工培养生物膜、沉积物和水)模拟自然条件下铅和镉的分布特征。研究结果显示:重金属进入到水环境之后,其浓度持续下降,Langmu ir动力学方程可以很好地描述其在水相中的变化规律。重金属在沉积物和生物膜中富集。生物膜对外界金属浓度变化的灵敏度高于沉积物。当水相中的重金属浓度降低到一个较小值时,生物膜中富集的重金属会被部分释放出来,通过水相,最终转移到沉积物中。沉积物对重金属的富集作用持续时间较长,富集速率较慢。水相、生物膜、沉积物三者共同作用,影响着重金属在自然水体中的分布规律。不同金属的分布过程变化趋势相似,但也存在一定差异。