Coastal mudflats as reservoirs of extracellular antibiotic resistance genes: Studies in Eastern China

Despite coastal mudflats serving as essential ecological zones interconnecting terrestrial/freshwater and marine systems, little is known about the profiles of antibiotic resistance genes (ARGs) in this area. In this study, characteristics of typical ARGs, involving both intracellular (iARGs) and extracellular ARGs (eARGs) at different physical states, were explored in over 1000 km of coastal mudflats in Eastern China. Results indicated the presence of iARGs and eARGs at states of both freely present or attached by particles. The abundance of eARGs was significantly higher than that of iARGs (87.3% vs 12.7%), and their dominance was more significant than those in other habitats (52.7%-76.3%). ARG abundance, especially for eARGs, showed an increasing trend (p < 0.05) from southern (Nantong) to northern (Lianyungang) coastal mudflats. Higher salinity facilitated the transformation from iARGs to eARGs, and smaller soil particle size was conducive to the persistence of eARGs in northern coastal mudflats. This study addresses the neglected function of coastal mudflats as eARGs reservoirs.     

Evaluation of the secondary structures of protein in the extracellular polymeric substances extracted from activated sludge by different methods

The changes of protein secondary structures in the extracellular polymeric substances(EPS) extracted from activated sludge by four different methods were studied by analyzing the amide I region(1700–1600 cm-1) of the Fourier transform infrared spectra and model protein test. The results showed the molecular weight distribution of organic matter extracted by centrifugation, heating and cation exchange resin(CER) was similar, while the EPS extracted by centrifugation(Control) and CER had similar fluorescent organic matter. The protein secondary structures of extracted EPS by the four methods were different. The similarities of protein secondary structures between the EPS extracted by CER with the Control were the highest among the four extracted EPS. Although the EPS yield extracted by formaldehyde + NaOH method were the highest, its protein secondary structures had the lowest similarity with those extracted by the Control. Additionally, the effects of centrifugation and CER extraction on the secondary structures of bovine serum albumin were also lower than that of other extraction processes. CER enables the second maximum extraction of EPS and maximum retention of the original secondary structure of proteins.     

The role of extracellular organic matter on the cyanobacteria ultrafiltration process

The membrane fouling caused by extracellular organic matter (EOM) and algal cells and organic matter removal of two typical cyanobacteria (M. aeruginosa and Pseudoanabaena sp.) during ultrafiltration (UF) process were studied in this work. The results showed that EOM had a broad molecular weight (Mw) distribution and the irreversible membrane fouling was basically caused by EOM. Moreover, humic acid and microbial metabolites were major components of EOM of two typical cyanobacteria. Since EOM could fill the voids of cake layers formed by the algal cells, EOM and algal cells played synergistic roles in membrane fouling. Fourier transform infrared spectroscopy analysis indicated that the CH₂ and CH₃ chemical bonds may play an important role in membrane fouling caused by EOM. Interestingly, the cake layer formed by the algal cells could trap the organic matter produced by algae and alleviate some irreversible membrane fouling. The results also showed that although the cake layer formed by the algal cells cause severe permeate flux decline, it could play a double interception role with UF membrane and increase organic matter removal efficiency. Therefore, when using UF to treat algae-laden water, the balance of membrane fouling and organic matter removal should be considered to meet the needs of practical applications.     

Stimulation of human amniotic fluid cell proliferation and colony formation by cell plating on a naturally produced extracellular matrix

Human amniotic fluid cells exhibit a higher cloning efficiency and rate of cell proliferation when maintained on dishes coated with a naturally produced extracellular matrix (ECM) in comparison with the regular tissue culture plastic. In 22 out of 31 amniotic fluid samples there was by plating the cells on ECM a 2–6 fold increase in number and size of colonies and in the cell density per colony as detected by actual staining and viewing of each colony. These effects yielded, in 21 of 41 additional samples, a reduction ranging from 2–8 days, in the culture time elapsing between amniocentesis and the first harvesting of cells for chromosomal analysis. An even greater effect was obtained with primary cells that failed to attach to plastic surfaces and stayed floating in the medium but did attach and proliferate when seeded on ECM. Cells that were left firmly attached to ECM after the first trypsinization and harvesting of cells for chromosomal analysis yielded colonies ready for second karyotyping in less than half the time required for cells maintained on plastic. Studies with secondary cultures of human amniotic fluid cells have demonstrated a 5–10 fold decrease in serum requirement of cells cultured on ECM as compared with plastic. Addition of fibroblast growth factor (FGF) to the cultures further potentiated the effects of ECM. The ECM induced stimulation of cell attachment and proliferation was not associated with any chromosomal anomalies, nor did it interfere with the handling procedure. ECM coated dishes may be useful to reduce the time interval between amniocentesis and diagnosis, in particular when the amniotic cells exhibit an exceedingly slow rate of proliferation on plastic or when large quantities of cells are required for enzymatic studies.     

Insights into the conversion of dissolved organic phosphorus favors algal bloom,arsenate biotransformation and microcystins release of Microcystis aeruginosa

Little information is available on influences of the conversion of dissolved organic phosphorus(DOP) to inorganic phosphorus(IP) on algal growth and subsequent behaviors of arsenate(As(V)) in Microcystis aeruginosa(M. aeruginosa). In this study, the influences factors on the conversion of three typical DOP types including adenosine-5-triphosphate disodium salt(ATP), β-glycerophosphate sodium(βP) and D-glucose-6-phosphate disodium salt(GP)were investigated under different extracellular polymeric ...     

Effect of 2-butenal manufacture wastewater to methanogenic activity and microbial community

The inhibition ratio sharply increased with the increasing COD. The absorbance of UV-vis at 420 nm showed a linear correlation with the SMA. The molecular structure of EPS has changed when COD was 9585 mg/L. Illumina Miseq sequencing was employed to reveal the microbial composition. The synthesis of 2-butenal, which is a vital raw material for the production of sorbic acid as a food preservative, generates some toxic by-products, so it is urgent to seek better detoxification strategies for the treatment of 2-butenal manufacture wastewater. In this study, batch experiments were carried out to investigate the inhibition effect of wastewater on the methanogenic activity. To understand the wastewater toxicity to anaerobic granular sludge, variations of the specific methanogenic activity (SMA) and extracellular polymeric substance (EPS) constituents at various wastewater CODs were investigated. Ultraviolet-visible (UV-vis) spectra and Fourier transform infrared (FT-IR) spectra were employed to analyze the structure of the EPS. The results showed that the inhibitory ratio of 2-butenal manufacture wastewater was less than 8.4% on the anaerobic granular sludge when the CODs were less than 959 mg/L. However, the inhibitory ratio increased from 36.4% to 93.6% when CODs increased from 2396 mg/L to 9585 mg/L, with the SMA decreasing from 39.1 mL CH₄/(gVSS·d) to 3.2 mL CH₄/(gVSS·d). The diversity of the microbial community under various CODs was researched by Illumina 16S rRNA Miseq sequencing and the results demonstrated that Proteiniphilum、Petrimonas and Syntrophobacter were the dominant bacteria genera in all sample. Regarding archaea, Methanobacterium was the most dominated archaea genera, followed by the Methanosaeta group in all samples. Moreover, the bacterial communities had changed obviously with increasing CODs, which indicated high CODs played a negative impact on the richness and diversity of bacterial community in the sludge samples.     

Coarsening of extracellularly biosynthesized cadmium crystal particles induced by thioacetamide in solution

A novel coarsening route for extracellularly biosynthesized cadmium nanocrystals was investigated for the first time. In this process, the white rot fungus Coriolus versicolor was employed to take up cadmium ions and synthesize extracellular cadmium crystal particles. The coarsening of the particles was induced by thioacetamide under certain conditions. Scanning electron microscopy showed that the formed cadmium crystal particles were coarsened from about 100 nm to 2-3 μm. The corresponding energy-dispersive X-ray spectra confirmed the presence of proteins in the particles. The maximum removal efficiency of Cd(II) increased from 17% to 87%, and the corresponding sorption capacity of biomass increased from 4 to 24 mg g⁻¹ with the completion of the coarsening process. The properties of the coarsened particles were also examined using X-ray diffraction (XRD) and transmission electron microscopy (TEM). XRD analysis of fungal mycelial pellets embedded with the coarsened particles confirmed the formation of cubic crystalline cadmium sulfide particles. The TEM results suggest that the coarsened particles were composed of clusters of several smaller particles. The changes in the functional groups on the biomass surface were studied through Fourier transform infrared spectroscopy. Based on the results above, a possible mechanism for the formation and coarsening of cadmium crystal particle is also discussed.     

活性污泥胞外聚合物中蛋白质的探针分析方法

应用微相吸附-光谱修正技术(Microsurface adsorption-spectral correction,简称MSASC),利用刚果红(Congo Red,简称CR)探针测定了活性污泥胞外聚合物(Extracellular polymeric substances,简称EPS)中蛋白质(Protein)的含量,与常用的Folin酚法比较,结果一致,蛋白加标回收率为94.5%～107.1%,相对标准偏差(RSD)＜4.5%.对反应机理的分析和产物结合常数的计算结果表明,在pH 2.21时CR-Protein作用符合Langmuir单分子层吸附理论,产物的结合比79∶1,结合常数K=2.17×106,聚集形式为Protein·CR79,摩尔吸收系数ε664r=8.50×105L·mol-1·cm-1.     

Characterisation of an extracellular polysaccharide produced by Bacillus mucilaginosus MY6-2 and its application in metal biosorption

A novel extracellular bioflocculant from Bacillus mucilaginosus MY6-2 was obtained by alcohol precipitation, dialysis and lyophilisation. The chemical analysis indicated that the bioflocculant was mainly composed of extracellular polysaccharide (named EPS MY6-2). Monosacchride composition result analysed by Gas chromatography indicated that EPS MY6-2 was mainly composed of Man, Glc, Gal, GlcA and GalA. The result of flocculating characteristics indicated that pH, EPS dosage and cation supplement had evident effect on flocculating activity of EPS MY6-2. The biosorption of iron ions from aqueous solution was tested with respect to pH, initial metal ion concentration and EPS dosage. The optimum biosorption conditions were as follows: pH 5.0, initial ion concentration from 200 to 800 mg/L and EPS dosage of 60 mg/L ion solution. The adsorption rates are maintained above 95% under these conditions. The adsorption data provided an excellent fit to both Langmuir and Freundlich isotherms, implying that the binding of the iron ions took place as a monolayer on the surface of the EPS.     

Influence of extracellular matrix on the proliferation of human amniotic fluid cells in vitro

Factors which influence the proliferation of human amniotic fluid cells in vitro have potential importance in reducing the time for prenatal diagnosis. Fibroblast growth factor (FGF) has been shown to be mitogenic for human amniotic fluid cells. The observation that cells which respond to FGF in vitro produce their own extracellular matrix (ECM), led to the use of an ECM as a substrate to assess proliferation. Pooled amniotic fluid cells maintained on an ECM prepared from bovine corneal endothelial cells demonstrated a significant increase in proliferation when compared with cells maintained on plastic substrate in the presence or absence of FGF. If FGF was added to cultures of amniotic fluid cells maintained on ECM, further increases in proliferation were noted compared with cells maintained on ECM in the absence of FGF. These results indicate that the substrate upon which amniotic fluid cells are maintained can have a profound influence on their proliferation.