Parametric and kinetic studies of activated sludge dewatering by cationic chitosan-like bioflocculant BF01314 produced from Citrobacter youngae.

Bacterial strains belonging to Citrobacter spp. were reported to produce polysaccharides consisting of N-acetylglucosamine and glucosamine like chitosan, with high flocculation activity. In this work, the flocculation dewatering performance of activated sludge conditioned by a novel cationic chitosan-like bioflocculant (BF) named BF01314, produced from Citrobacter youngae GTC 01314, was evaluated under the influences of flocculant dosage, pH, and temperature. At BF dosage as low as 0.5 kg/t DS, the sludge dewaterability was significantly enhanced in comparison to the raw (untreated) sludge, featuring well-flocculated characteristic (reduction in CST from 22.0 s to 9.4 s) and good sludge filterability with reduced resistance (reduction in SRF by one order from 7.42 × 1011 to 9.59 × 1010 m/kg) and increased compactness of sludge (increase in CSC from 15.2 to 23.2%). Besides, the BF demonstrated comparable high sludge dewatering performance within the pH range between 2 and 8, and temperature range between 25 °C and 80 °C. Comparison between the BF, the pristine chitosan and the commercial cationic copolymer MF 7861 demonstrated equivalent performance with enhanced dewaterability at the dosage between 2.0 and 3.0 kg/t DS. Besides, the BF demonstrated strong flocculation activity (>99%) when added to the sludge suspension using moderate to high flocculation speeds (100-200 rpm) with at least 3-min mixing time. The BF's reaction in sludge flocculation was best fitted with a pseudo first-order kinetic model. Electrostatic charge patching and polymer bridging mechanisms are believed to be the dominant mechanistic phenomena during the BF's sludge conditioning process (coagulation-flocculation).

Citrobacter braakii bacteremia-induced septic shock after colonoscopy preparation with polyethylene glycol in a critically ill patient: a case report.

BACKGROUND: Polyethylene glycol (PEG) is widely used for bowel cleaning in preparation for colonoscopy because of its safety. Septic shock after PEG preparation is an extremely rare complication. Herein, we describe a case of septic shock that occurred immediately after colonoscopy preparation with PEG. CASE PRESENTATION: A 75-year-old Japanese male who had previously developed diabetes after total pancreatectomy received PEG in preparation for colonoscopy. He had been admitted to the emergency intensive care unit 4 days earlier due to hematochezia presenting with shock. He ingested PEG to prepare for a colonoscopy examination, which was performed to identify the source of his bleeding over a 5-h period, but suddenly exhibited septic shock and markedly elevated procalcitonin levels. A blood culture subsequently revealed Citrobacter braakii. Immediate resuscitation and intensive care with appropriate antibiotics improved his condition. CONCLUSIONS: Clinicians should be aware of the possibility of deteriorating conditions after bowel preparation with PEG among severely ill patients with recent episodes of hemorrhagic shock.

Bacterial succession and degradative changes by biofilm on plastic medium for wastewater treatment.

Biofilms contain a diverse range of microorganisms and their varying extracellular polysaccharides. The present study has revealed biofilm succession associated with degradative effects on plastic (polypropylene) and contaminants in sludge. The wet weight of biofilm significantly (p < 0.05) increased; from 0.23 ± 0.01 to 0.44 ± 0.01 g. Similarly, the dry weight of the biofilm increased from 0.02 to 0.05 g. Significant reduction in pathogens (E. coli and feacal coliforms) by MPN technique (>80%) and in chemical parameters (decrease in COD, BOD5 of 73.32 and 69.94%) representing diminution of organic pollutants. Energy dispersive X-ray spectroscopy (EDS) of plastic revealed carbon and oxygen contents, further surface analysis of plastic by scanning electron microscopy (SEM) revealed emergence of profound bacterial growth on the surface. Fourier transform infrared (FTIR) spectroscopy conforms its biotransformation under aerobic conditions after 8 weeks. New peaks developed at the region 1050 and 969 cm(-1) indicating CO and CC bond formation. Thus plastic with 6 weeks old aerobic biofilm (free of pathogens, max. weight, and OD, efficient COD & BOD removal ability) is suggested to be maintained in fixed biofilm reactors for wastewater treatment.

Microbiological corrosion acceleration of N80 steel in shale gas field produced water containing Citrobacter amalonaticus at 60 °C.

Microbiologically influenced corrosion (MIC) is one of typical reasons leading to a lot of damage of pipeline steels in the shale gas environments. MIC behavior and mechanism studies are important towards to steel protection. In this work, Citrobacter amalonaticus, a corrosive bacterium, was isolated from a shale gas well of China. And the corrosion behavior of N80 steel caused by C. amalonaticus was studied in simulated shale gas produced water at 60 °C making use of weight loss, surface analysis, electrochemical impedance spectroscopy (EIS), anodic potentiostatic polarization measurements, and potentiodynamic polarization curves. Results demonstrate that C. amalonaticus could accelerate the uniform and localized corrosion rates of N80 steel at 60 °C with values of (0.221 ± 0.016) and (0.557 ± 0.062) mm/y, respectively. Experimental results suggested that the adsorption of an organic inhibitor film on steel surface caused the corrosion rates of abiotic specimens going down. However, the existence of C. amalonaticus inhibited the adsorption of organic inhibitor film. The adhesion and biofilm formation of C. amalonaticus contributed to steel corrosion acceleration. The nucleation and growth of metastable pitting were enhanced by C. amalonaticus, thus causing a severe localized corrosion.

Bacterial decolorization of black liquor in axenic and mixed condition and characterization of metabolites.

The pulping byproducts (black liquor) cause serious environmental problem due to its high pollution load. In order to search the degradability of black liquor, the potential bacterial strains Citrobacter freundii (FJ581026) and Citrobacter sp. (FJ581023) were applied in axenic and mixed condition. Results revealed that the mixed bacterial culture are more effective than axenic condition and can reduce 82% COD, 79% AOX, 79% color and 60% lignin after 144 h of incubation period. Additionally, the optimum activity of lignin degrading enzyme was noted at 96 h and characterized as manganese peroxidase (MnP) by SDS­PAGE analysis. Further, the HPLC analysis of control and bacterial degraded sample has shown the reduction as well as shifting of peaks compared to control indicating the degradation as well as transformation of compounds of black liquor. The comparative GC-MS analysis of control and degraded black liquor revealed that along with lignin fragment some chlorophenolic compounds 2,4,6-trichlorophenol, 2,3,4,5-tetrachlorophenol and pentachlorophenol were detected in black liquor degraded by axenic culture whereas these chlorophenolic compounds were completely absent in black liquor degraded by mixed bacterial culture. These chlorophenol inhibit the oxidative degradation which seems a major reason behind the low degradability of axenic degradation compared to mixed culture. The innovation of this aerobic treatment of alkaline black liquor opens additional possibilities for the better treatment of black liquor along with its metabolic product.

Toxic effect of high concentration of sonochemically synthesized polyvinylpyrrolidone-coated silver nanoparticles on Citrobacter sp. A1 and Enterococcus sp. C1.

BACKGROUND/PURPOSE: Currently, silver nanoparticles (AgNPs) have gained importance in various industrial applications. However, their impact upon release into the environment on microorganisms remains unclear. The aim of this study was to analyze the effect of polyvinylpyrrolidone-capped AgNPs synthesized in this laboratory on two bacterial strains isolated from the environment, Gram-negative Citrobacter sp. A1 and Gram-positive Enterococcus sp. C1. METHODS: Polyvinylpyrrolidone-capped AgNPs were synthesized by ultrasound-assisted chemical reduction. Characterization of the AgNPs involved UV-visible spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction, transmission electron microscopy, and energy dispersive X-ray spectroscopy. Citrobacter sp. A1 and Enterococcus sp. C1 were exposed to varying concentrations of AgNPs, and cell viability was determined. Scanning electron microscopy was performed to evaluate the morphological alteration of both species upon exposure to AgNPs at 1000 mg/L. RESULTS: The synthesized AgNPs were spherical in shape, with an average particle size of 15 nm. The AgNPs had different but prominent effects on either Citrobacter sp. A1 or Enterococcus sp. C1. At an AgNP concentration of 1000 mg/L, Citrobacter sp. A1 retained viability for 6 hours, while Enterococcus sp. C1 retained viability only for 3 hours. Citrobacter sp. A1 appeared to be more resistant to AgNPs than Enterococcus sp. C1. The cell wall of both strains was found to be morphologically altered at that concentration. CONCLUSION: Minute and spherical AgNPs significantly affected the viability of the two bacterial strains selected from the environment. Enterococcus sp. C1 was more vulnerable to AgNPs, probably due to its cell wall architecture and the absence of silver resistance-related genes.

Enzymically accelerated biomineralization of heavy metals: application to the removal of americium and plutonium from aqueous flows.

A biological process for the removal of heavy metals from the aqueous flows is described. Metals are precipitated on the surface of immobilized cells of a Citrobacter sp. as cell-bound metal phosphates. This uses phosphate liberated by the activity of a cell-bound phosphatase. Some radionuclides (e.g. 241americium) form metal phosphates readily; efficient removal of 241Am on a continuous basis is demonstrated. At low phosphatase activities, the efficiency of uranium removal correlates with enzyme activity. High phosphatase activities are not realised as an increase in metal removal, suggesting that chemical events become rate-limiting. Studies have suggested that maximal metal uptake occurs only after nucleation and the formation of precipitation foci. A model is presented to illustrate how nucleation and crystallization processes could enhance the removal of plutonium and neptunium from dilute solutions.

Enhanced biohydrogen production from corn stover by the combination of Clostridium cellulolyticum and hydrogen fermentation bacteria.

Hydrogen was produced from steam-exploded corn stover by using a combination of the cellulolytic bacterium Clostridium cellulolyticum and non-cellulolytic hydrogen-producing bacteria. The highest hydrogen yield of the co-culture system with C. cellulolyticum and Citrobacter amalonaticus reached 51.9 L H2/kg total solid (TS). The metabolites from the co-culture system were significantly different from those of the mono-culture systems. Formate, which inhibits the growth of C. cellulolyticum, could be consumed by the hydrogen-evolving bacteria, and transformed into hydrogen. Glucose and xylose were released from corn stover via hydrolysis by C. cellulolyticum and were quickly utilized in dark fermentation with the co-cultured hydrogen-producing bacteria. Because the hydrolysis of corn stover by C. cellulolyticum was much slower than the utilization of glucose and xylose by the hydrogen-evolving bacteria, the sugar concentrations were always maintained at low levels, which favored a high hydrogen molar yield.

Quorum sensing activity of Citrobacter amalonaticus L8A, a bacterium isolated from dental plaque.

Cell-cell communication is also known as quorum sensing (QS) that happens in the bacterial cells with the aim to regulate their genes expression in response to increased cell density. In this study, a bacterium (L8A) isolated from dental plaque biofilm was identified as Citrobacter amalonaticus by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). Its N-acylhomoserine-lactone (AHL) production was screened by using two types of AHL biosensors namely Chromobacterium violaceum CV026 and Escherichia coli [pSB401]. Citrobacter amalonaticus strain L8A was identified and confirmed producing numerous types of AHL namely N-butyryl-L-homoserine lactone (C4-HSL), N-hexanoyl-L-homoserine lactone (C6-HSL), N-octanoyl-L-homoserine lactone (C8-HSL) and N-hexadecanoyl-L-homoserine lactone (C16-HSL). We performed the whole genome sequence analysis of this oral isolate where its genome sequence reveals the presence of QS signal synthase gene and our work will pave the ways to study the function of the related QS genes in this bacterium.

Biochemical characterization of a bifunctional acetaldehyde-alcohol dehydrogenase purified from a facultative anaerobic bacterium Citrobacter sp. S-77.

Acetaldehyde-alcohol dehydrogenase (ADHE) is a bifunctional enzyme consisting of two domains of an N-terminal acetaldehyde dehydrogenase (ALDH) and a C-terminal alcohol dehydrogenase (ADH). The enzyme is known to be important in the cellular alcohol metabolism. However, the role of coenzyme A-acylating ADHE responsible for ethanol production from acetyl-CoA remains uncertain. Here, we present the purification and biochemical characterization of an ADHE from Citrobacter sp. S-77 (ADHE(S77)). Interestingly, the ADHE(S77) was unable to be solubilized from membrane with detergents either 1% Triton X-100 or 1% Sulfobetaine 3-12. However, the enzyme was easily dissociated from membrane by high-salt buffers containing either 1.0 M NaCl or (NH(4))(2)SO(4) without detergents. The molecular weight of a native protein was estimated as approximately 400 kDa, consisting of four identical subunits of 96.3 kDa. Based on the specific activity and kinetic analysis, the ADHES77 tended to have catalytic reaction towards acetaldehyde elimination rather than acetaldehyde formation. Our experimental observation suggests that the ADHES77 may play a pivotal role in modulating intracellular acetaldehyde concentration.

Crystallization and crystal data on tyrosine phenol-lyase.

Crystals of the apoenzyme of tyrosine phenol-lyase (EC 4.1.99.2), a pyridoxal 5'-phosphate-dependent enzyme from Citrobacter intermedius, have been grown by vapor diffusion of an ammonium sulfate solution to a protein solution. The crystals belong to space group P2(1)2(1)2, with dimensions of a = 75.5 A, b = 138.4 A and c = 94.1 A and diffract up to 2.7 A resolution. The asymmetric unit contains one half of the enzyme tetrameric molecule. Two heavy-atom derivatives of the crystals have been obtained.

Biosorption of lead, cadmium, and zinc by Citrobacter strain MCM B-181: characterization studies.

The biosorption process for removal of lead, cadmium, and zinc by Citrobacter strain MCM B-181, a laboratory isolate, was characterized. Effects of environmental factors and growth conditions on metal uptake capacity were studied. Pretreatment of biomass with chemical agents increased cadmium sorption efficiency; however, there was no significant enhancement in lead and zinc sorption capacity. Metal sorption by Citrobacter strain MCM B-181 was found to be influenced by the pH of the solution, initial metal concentration, biomass concentration, and type of growth medium. The metal sorption process was not affected by the age of the culture or change in temperature. Equilibrium metal sorption was found to fit the Langmuir adsorption model. Kinetic studies showed that metal uptake by Citrobacter strain MCM B-181 was a fast process, requiring <20 min to achieve >90% adsorption efficiency. The presence of cations reduced lead, zinc, and cadmium sorption to the extent of 11. 8%, 84.3%, and 33.4%, respectively. When biomass was exposed to multimetal solutions, metals were adsorbed in the order Co2+ < Ni2+ < Cd2+ < Cu2+ < Zn2+ < Pb2+. Among various anions tested, only phosphate and citrate were found to hamper metal sorption capacity of cells. Biosorbent beads prepared by immobilizing the Citrobacter biomass in polysulfone matrix exhibited high metal loading capacities. A new mathematical model used for batch kinetic studies was found to be highly useful in prediction of experimentally obtained metal concentration profiles as a function of time. Metal desorption studies indicated that Citrobacter beads could, in principle, be regenerated and reused in adsorption-desorption cycles. In an expanded scale trial, biosorbent beads were found to be useful in removal/recovery of metals such as lead from industrial wastewaters.

Contamination of drinking water by coliforms from biofilms grown on rubber-coated valves.

In water samples from drinking water distribution systems, coliform bacteria (predominantly Citrobacter species) were repeatedly detected. Disinfection and flushing of the systems did not erase the problem. The pattern of the coliform occurrences indicated contamination originating from biofilms. After inspection of internal surfaces of the systems, no significant biofilm growth was observed on pipe surfaces, but in a number of cases, visible biofilms were detected on rubber-coated valves which harboured the same coliform species as those found in the drinking water samples. In these cases, the rubber-coated valves seemed to act as point sources for the contamination of water.

Assessment of macroporous polystyrene-based polymers for the immobilization of Citrobacter freundii.

POLYHIPE polymers represent a novel immobilization support possessing many desirable characteristics. These materials have been used to immobilize Citrobacter freundii in order to develop a continuous bioconversion system for the anaerobic production of trimethylene glycol from glycerol. Several immobilization methods were employed, of which the most successful involved culturing the organism with POLYHIPE in shake-flasks, transferring the contents into a column, and growing the cells in situ within the column. Cell activity was determined by the amount of TMG produced from a buffered glycerol solution. The latter was passed through the columns by either a recirculation system or the superior single-pass method. The optimal system was scaled up and showed potential for further large-scale investigations. Cell lifetime in the column could be improved by supplying the cells with diluted growth medium in place of the buffer.

[Enzymatic synthesis of 3,4-dihydroxyphenyl-L-alanine by free and immobilized Citrobacter freundii cells]. / Fermentativnyi sintez 3,4-dioksifenil-L-alanina svobodnymi i immobilizovannymi kletkami Citrobacter freundii.

The Citrobacter freundii 62 cells immobilized in PAAG and possessing the tyrosine-phenol-lyase (TPL) activity catalyse the synthesis of 3,4-dihydroxyphenyl-L-alanine (DOPA) from pyrocatechol and ammonium pyruvate. The synthesis of DOPA was studied using both free and immobilized bacterial cells. When the concentration of pyrocatechol is over 0.1 M the TPL activity of the cells is inhibited. The concentration of pyrocatechol can be increased up to 0.3 M by using an equimolar mixture of pyrocatechol and boric acid. The addition of ascorbic acid as an antioxidant results in a lower TPL activity of both free and immobilized bacterial cells.

[Immobilization of Citrobacter L-asparaginase in polyacrylamide gel]. / Immobilizatsiia L-asparaginazy iz Citrobacter v poliakrilamidnom gele.

Bacterial L-asparaginase, immobilized on polyacrylamide gel, exhibited higher stability to denaturation and to the effect of a proteolytic enzyme. The immobilized enzyme exhibited the pH optimum of activity displaced by one pH unit to the acid side as compared with the free enzyme. The apparent Km value was approximately 200-fold higher as compared with the free L-asparaginase. The immobilized asparaginase hydrolyzed both L- and D-asparagine isomers but the free enzyme was highly stereospecific.

Selective oxidation of glycerol to 1,3-dihydroxyacetone by covalently immobilized glycerol dehydrogenases with higher stability and lower product inhibition.

Glycerol dehydrogenase (GlyDH) catalyzes the regioselective oxidation of glycerol to yield 1,3-dihydroxyacetone (DHA); an important building block in chemical industry. Three recombinant GlyDHs from Geobacillus stearothermophilus, from Citrobacter braakii and from Cellulomonas sp. were stabilized by covalent immobilization. The highest activity recoveries (40-50%) of the insoluble preparations were obtained by immobilizing these enzymes in presence of polyethylene glycol (PEG). Noteworthy, these immobilized preparations were more stable and less inhibited by DHA than their soluble counterparts. In particular, GlyDH from G.stearothermophilus immobilized on agarose activated with both amine and glyoxyl groups and crosslinked with dextran aldehyde was 3.7-fold less inhibited by DHA than its soluble form and retained 100% of its initial activity after 18h of incubation at 65°C and pH 7. This is one of the few examples where the same immobilization protocol has minimized enzyme product inhibition and maximized thermal stability.

Surfactant-modified fatty acid composition of Citrobacter sp. SA01 and its effect on phenanthrene transmembrane transport.

The effects of the surfactants, Tween 80 and sodium dodecyl benzene sulfonate (SDBS) on a membrane's fatty acid composition and the transmembrane transport of phenanthrene were investigated. The results indicated that both surfactants could modify the composition of fatty acids of Citrobacter sp. Strain SA01 cells, 50 mg L(-1) of both surfactants changed the composition of the fatty acids the most, increasing the amount of unsaturated fatty acids. The comparison of fatty acid profiles with diphenylhexatriene fluorescence anisotropy, a probe for plasma membrane fluidity, suggested that an increased amount of unsaturated fatty acids corresponded to greater membrane fluidity. In addition, increased unsaturated fatty acids promoted phenanthrene to partition from the extracellular matrix to cell debris, which increased reverse partitioning from the cell debris to the cytochylema. The results of this study were expected in that the addition of a surfactant is a simple and effective method for accelerating the rate-limiting step of transmembrane transport of hydrophobic organic compounds (HOCs) in bioremediation.

The microbial flora associated with oral carcinomas.

OBJECTIVE: Changes in the microbial flora on the oral mucosa after cancerous alteration may lead to both local and systemic infections. In this study, we assessed the microbial flora associated with the surfaces of oral squamous cell carcinoma. A comparative evaluation of these microbial contents was made with that of the contralateral healthy mucosa and control (healthy) mucosa. We also assessed the microbial flora from the saliva culture in subjects with oral squamous cell carcinoma and healthy controls. METHOD AND MATERIALS: The case control study was made up of 30 subjects with oral squamous cell carcinoma as the study group; 30 healthy age-, sex-, habit-, and dentition-matched subjects served as the control group. In the study group, microbial samples were collected from the carcinoma site, contralateral healthy mucosa, and saliva, whereas in the control group, samples were collected from the healthy mucosa and saliva. These samples were stored on ice and subsequently transported to the laboratory in 2 mL of thioglycollate transport media, where the microbial cultures were carried out. RESULTS: Oral squamous cell carcinoma sites harbor significantly more microbial flora (bacteria and yeasts) compared to those of healthy mucosa (control group). The microbial flora predominantly isolated from the carcinoma site were Streptococcus species, Staphylococcus species, Moraxella species, Enterococcus feacalis, Aerobic spore bearers, Klebsiella species, Citrobacter species, Proteus species, Pseudomonas species, and Candida albicans. The median number of colony forming units (CFU)/mL at carcinoma sites (3.85 x 105 CFU/mL) was significantly higher than that of the healthy mucosa (0.571 x 105 CFU/mL; P = .0000, Wilcoxon nonparametric test). Similarly, in saliva of carcinoma subjects, the median number of CFU/mL (2.408 x 105 CFU/mL) was significantly higher than that of saliva in control subjects (0.78 x 105 CFU/mL; P = .0000, Wilcoxon nonparametric test). CONCLUSION: The present study clearly indicates that the subjects with oral squamous cell carcinoma harbor significantly more microbial flora. Emphasis has to be given to preventing microbial flora in the oral cavity and treating these patients with appropriate antimicrobial agents, thus reducing their morbidity.