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Fomites are inanimate objects that are capable of absorbing, harboring and transmitting infectious microorganisms. Paper currency, an exchangeable fomite, is constantly subjected to contamination. This study investigated bacterial contamination on the Saudi one Riyal paper note. Two hundred one Riyal bills (100 4th version and 100 5th version) were collected in the city of Jeddah for examination. The notes were examined for bacterial contamination using blood agar and MacConkey agar. Eighty-eight percent of the notes were contaminated with mixed (>or=2 types) bacterial growth. All 4th version notes had mixed bacterial growth: gram-positive bacilli (79%), coagulase-negative staphylococci (75%), Staphylococcus aureus (38%), Klebsiella spp (21%), Pseudomonas spp (19%), Escherichia coli (9%), viridans group streptococci (VGS) (8%), and non-hemolytic streptococci (4%). Seventy-six percent of the newer 5th version notes had mixed bacterial growth: gram-positive bacilli (68%), coagulase-negative staphylococci (64%), S. aureus (13%), Klebsiella spp (9%), Pseudomonas spp (5%), E. coli (2%) and VGS (2%). These results indicate the Saudi one Riyal paper note is commonly contaminated with bacteria, and may act as a vehicle for the transmission of potentially pathogenic bacteria. The more the bill had been handled the greater the contamination was.

Methanol-oxidizing bacteria may play an important role in the development and use of biological treatment systems for the removal of methanol from industrial effluents. Optimization of methanol degradation potential in such systems is contingent on availability of nutrients, such as nitrogen, in the most favorable form and concentration. To that end, this study examined the variation in growth, methanol degradation, and bacterial diversity of two mixed methylotrophic cultures that were provided nitrogen either as ammonium or nitrate and in three different concentrations. Methanol-degrading cultures were enriched from biofilms sampled at a pulp and paper mill and grown in liquid batch culture with methanol as the only carbon source and either ammonium or nitrate as the only added nitrogen source. Results indicate that growth and methanol removal of the mixed cultures increase directly with increased nitrogen, added in either form. However, methanol removal and bacterial diversity, as observed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) methods, were higher when using nitrate as the nitrogen source for enrichment and growth, rather than ammonium. Based on results described here, nitrate may potentially be a better nitrogen source when enriching or working with mixed methylotrophic cultures, and possibly more effective when used as a nutrient addition to biofilters.

There is an urgent need for cheap point-of-use methods to purify drinking water. We describe a method to deactivate pathogenic bacteria by percolation through a paper sheet containing silver nanoparticles. The silver nanoparticles are deposited by the in situ reduction of silver nitrate on the cellulose fibers of an absorbent blotting paper sheet. The aim is to achieve inactivation of bacteria during percolation through the sheet, rather than removal of bacteria from the effluent by filtration. The silver-nanoparticle containing (AgNP) papers were tested for performance in the laboratory with respect to bacteria inactivation and silver leaching as suspensions of bacteria percolated through the paper. The AgNP sheets exhibited antibacterial properties toward suspensions of Escherichia coli and Enterococcus faecalis, with log reduction values in the effluent of over log 6 and log 3, respectively. The silver loss from the AgNP sheets was minimal, with values under 0.1 ppm (the current US EPA and WHO limit for silver in drinking water). These results show promise that percolation of bacterially contaminated water through paper embedded with silver nanoparticles could be an effective emergency water treatment.

Four different bacterial strains were isolated from pulp and paper mill sludge in which one alkalotolerant isolate (LP1) having higher capability to remove color and lignin, was identified as Bacillus sp. by 16S RNA sequencing. Optimization of process parameters for decolorization was initially performed to select growth factors which were further substantiated by Taguchi approach in which seven factors, % carbon, % black liquor, duration, pH, temperature, stirring and inoculum size, at two levels, applying L-8 orthogonal array were taken. Maximum color was removed at pH 8, temperature 35°C, stirring 200 rpm, sucrose (2.5%), 48 h, 5% (w/v) inoculum size and 10% black liquor. After optimization 2-fold increase in color and lignin removal from 25-69% and 28-53%, respectively, indicated significance of Taguchi approach in decolorization and delignification of lignin in pulp and paper mill effluent. Enzymes involved in the process of decolorization of effluent were found to be xylanase (54 U/ml) and manganese peroxidase (28 U/ml). Treated effluent was also evaluated for toxicity by Comet assay using Saccharomyces cerevisiae MTCC 36 as model organism, which indicated 58% reduction after treatment by bacterium.

Herein, we report the development of a novel, inexpensive, and portable filter-paper-based strip biosensor for the detection of bacterial quorum sensing signaling molecules, N-acylhomoserine lactones (AHLs). AHLs are generally employed by Gram-negative bacteria for their cell-cell communication to control expression of specialized genes, such as those involved in biofilm formation and production of virulence factors, in a population-density-dependent manner. First, a bacterial cell-based sensing system employing components of AHL-mediated QS regulatory system as recognition elements and beta-galactosidase as the reporter protein was designed and developed. The bacterial-sensing cells were then liquid-dried on strips of filter paper. beta-Galactosidase as the reporter allows for the visual monitoring of the analyte-induced signal when a colorimetric method of detection is applied. The paper strip biosensor was able to detect low AHL concentrations down to 1 x 10(-8) M. Furthermore, it was successfully applied to the detection of AHLs in physiological samples, such as saliva. The filter-paper-based sensing strips could provide reproducible results upon storage at 4 degrees C for at least 3 months. In conclusion, a filter-paper-based strip biosensor was developed that allows for visual, fast, and convenient detection of AHLs in a dose-dependent manner in a test sample. In addition, it does not require expensive equipment or trained personnel and allows ease of transportation and storage. Therefore, we envision that this biosensor will serve as a simple and economical portable field kit for on-site monitoring of AHL in a variety of clinical and environmental samples.

Formation of biofilms causes severe problems in paper machines, and hence financial costs. It would be preferable to prevent attachment of the primary-colonizing bacteria than to control the growth of secondary communities, which are sheltered by exopolysaccharide slime layers. We have therefore investigated the early succession of paper-machine biofilms by incubating stainless-steel test coupons in the process water-flow lines in two paper machines operating in slightly alkaline conditions in temperatures (45 and 49 degrees C) supporting thermophilic microbes. Microbial succession was profiled using length heterogeneity analysis of PCR-amplified 16S rRNA genes (LH-PCR) and linking the sequence data of the created 16S rRNA gene libraries to the dominant LH-PCR peaks. Although the bacterial fingerprints obtained from the attached surface communities varied slightly in different samples, the biomarker signals of the dominating primary-colonizing bacterial groups remained high over time in each paper machine. Most of the 16S rRNA gene copies in the early biofilms were assigned to the genera Rhodobacter, Tepidimonas, and Cloacibacterium. The dominance of these sequence types decreased in the developing biofilms. Finally, as phylogenetically identical primary-colonizers were detected in the two different paper mills, the machines evidently had similar environmental conditions for bacterial growth and potentially a common source of contamination.

Forty-one bacterial strains isolated from the headbox water of two machines in a Canadian paper mill were associated with the genera Asticcacaulis, Acidovorax, Bacillus, Exiguobacterium, Hydrogenophaga, Pseudomonas, Pseudoxanthomonas, Staphylococcus, Stenotrophomonas based on the sequence of their 16S rRNA genes. The metabolic profile of these strains were determined using Biolog EcoPlate, and the bacteria were divided into four metabolic groups. Metabolic profiles of the bacterial communities colonizing the headbox water of two paper machines was also determined weekly over a 1 year period. The only compound that was not reduced by the bacterial community was 2-hydroxybenzoic acid. Utilization frequency of the other carbon sources in the Biolog EcoPlate ranged from 3 to 100%. The metabolic profiles of the bacterial community did not vary considerably between the two paper machines. However, the metabolic profile varied among the sampling dates.

A total of 179 non-spore-forming bacteria aerobically growing on Nutrient Agar, Plate Count Agar or in specific enrichment conditions for salmonella, campylobacteria, listeria, yersinia or staphylococci, were isolated from 16 untreated paper mill pulps. After phenotypical screening the isolates were characterised by automated ribotyping and partial sequencing of the 16S rRNA gene. They could be divided into seven taxonomical classes representing 63 taxa (species): actinobacteria (11 species), bacilli (7), flavobacteria (3) alphaproteobacteria (10), betaproteobacteria (5), gammaproteobacteria (25) and sphingobacteria (2). Most of the gammaproteobacteria were enterobacteria, mainly species of the genera Enterobacter (7 species, 7 samples/3 mills) and Klebsiella (5 species, 6 samples/3 mills). Other commonly occurring bacteria were most closely related to Microbacterium barkeri (7 samples/3 mills), Cloacibacterium normanense (6 samples/2 mills), Pseudoxanthomonas taiwanensis (5 samples/2 mills) and Sphingobacterium composti (5 samples/1 mill). Sporadic isolates of Listeria innocua, L. monocytogenes, Enterococcus casseliflavus and Staphylococcus warneri were detected, from which only L. monocytogenes is considered to be a food pathogen. No isolates of the genera Campylobacter, Salmonella or Yersinia were detected. The detected bacteria may be harmful in process control, but the load of food pathogens with recycled fibres to paper machines is insignificant. Faecal contamination of the pulp samples was not indicated.

Efficient utilization of carbon inputs is critical to the economic viability of the current forest products sector. Input carbon losses occur in various locations within a pulp mill, including losses as volatile organics and wastewater. Opportunities exist to capture this carbon in the form of value-added products such as biodegradable polymers. Waste-activated sludge from a pulp mill wastewater facility was enriched for 80 days for a methanol-utilizing consortium with the goal of using this consortium to produce biopolymers from methanol-rich pulp mill waste streams. Five enrichment conditions were utilized: three high-methanol streams from the kraft mill foul condensate system, one methanol-amended stream from the mill wastewater plant, and one methanol-only enrichment. Enrichment reactors were operated aerobically in sequencing batch mode at neutral pH and 25 degrees C with a hydraulic residence time and a solids retention time of 4 days. Non-enriched waste activated sludge did not consume methanol or reduce chemical oxygen demand. With enrichment, however, the chemical oxygen demand reduction over 24-h feed/decant cycles ranged from 79 to 89%, and methanol concentrations dropped below method detection limits. Neither the non-enriched waste-activated sludge nor any of the enrichment cultures accumulated polyhydroxyalkanoates (PHAs) under conditions of nitrogen sufficiency. Similarly, the non-enriched waste activated sludge did not accumulate PHAs under nitrogen-limited conditions. By contrast, enriched cultures accumulated PHAs to nearly 14% on a dry weight basis under nitrogen-limited conditions. This indicates that selectively enriched pulp mill waste activated sludge can serve as an inoculum for PHA production from methanol-rich pulp mill effluents.

The authors assessed the potential of Nigerian currency notes to act as environmental vehicles for the transmission of pathogenic parasites and bacteria. Currency notes obtained from four major cities in Nigeria were evaluated according to standard techniques. Fifty-four (21.6 percent [95 percent CI: 16.50-26.70]) of the first batch of 250 notes, which underwent parasitological analysis, were contaminated with enteric parasites; 133 (53.2 percent [95 percent CI: 47.02-59.39]) of the second batch of 250 notes, which underwent bacteriological analysis, were found to be contaminated with bacteria. Parasites that were isolated from the notes included Ascaris lumbricoides (8.0 percent), Enterobius vermicularis (6.8 percent), Trichuris trichiura (2.8 percent), and Taenia species (4.0 percent). Bacteria that were isolated were Streptococcus species (21.6 percent), Staphylococcus species (12.8 percent), Escherichia coli (13.2 percent), and Bacillus species (5.6 percent). Among dirty/mutilated currency notes, parasite contamination and bacterial contamination were both significantly (p < .05) more pervasive (30.6 percent and 73.8 percent, respectively) than they were among clean and mint currency notes. Lower-denomination notes were more likely to be contaminated than were higher-denomination notes, although the difference was not statistically significant (p > .05). Parasite contamination and bacteria contamination were both most frequent in notes obtained from butchers and beggars. These results suggest that currency notes may be contaminated, especially with bacteria and enteric parasites, and may serve as sources of infection. Personal hygiene to reduce risk of infection is recommended.

Paper mill sludge is a solid waste material composed of pulp residues and ash generated from pulping and paper making process. The carbohydrate portion of the sludges from Kraft/Recycle paper mill has chemical and physical characteristics similar to those of commercial wood pulp. Because of its high carbohydrate content and well-dispersed structure, the sludge can be biologically converted to value-added products without pretreatment. In bioconversion of solid feedstock such as paper mill sludge, a certain amount of water must be present to attain fluidity. In this study, hemicellulose pre-hydrolysate, in place of water, was added to the sludge to increase the concentration of the final product. Pre-hydrolysate was obtained by hot-water treatment of pine wood in which the total sugar concentration reached 4 wt.%. The mixture was processed by simultaneous saccharification and fermentation (SSF) using enzymes (cellulase and pectinase) and Lactobacillus rhamnosus (ATCC-10863). Pectinase was added to hydrolyze mannose oligomers in the pre-hydrolysate to monomers. During the SSF of the mixture, calcium carbonate in the paper sludge acted as a buffer, yielding calcium lactate as the final product. External pH control was unnecessary due to the buffer action of calcium carbonate that maintained the pH near optimum for the SSF. The lactic acid yield in the range of 80-90 % of the theoretical maximum was obtained. Use of the mixed feed of pre-hydrolysate and pulp mill sludges in the SSF raised the product concentration to 60 g of lactate/L.

This work comprises the use of a multi-analytical approach combined with microbiological studies to characterize six paper samples, containing foxing stains, from the 20th century, regarding their cellulose matrix, fillers, and sizing materials, and to evaluate possible paper degradation that might have occurred during the foxing stains. Photography under different illuminations and optical microscopy were used for morphological characterization of the paper samples and foxing stains. Scanning electron microscopy coupled energy dispersive spectroscopy (SEM-EDS) was of particular importance for defining the presence of fiber disorder and disruption on the surface of some of the stains, and localized accumulations of mineral-like particles on the surface of others. SEM-EDS, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FT-IR), and energy dispersive X-ray fluorescence (EDXRF) were used for the identification of mineral fillers, whereas sizing agents were analyzed using ATR-FT-IR. EDXRF results showed that no differences, within the standard deviation, were found in iron and copper contents between the foxed and unfoxed areas. Fungi belonging to the genus Penicillium spp. were found in all the paper samples. Unfoxed areas presented lower contamination than the foxed areas.

Aims: To isolate aerobic mesophilic bacilli and thermophilic bacteria from different paper mill samples and to evaluate their potential harmfulness., Methods and Results: A total of 109 mesophilic and 68 thermophilic isolates were purified and characterized by automated ribotyping and partial 16S rDNA sequencing. The mesophilic isolates belonged to the genera Bacillus (13 taxa), Brevibacillus (three taxa) and Paenibacillus (five taxa). The thermophilic bacteria represented seven taxa of Bacillus, Geobacillus or Paenibacillus, four of proteobacteria and one of actinobacteria. The most frequently occurring bacteria were Bacillus cereus, B. licheniformis, Pseudoxanthomonas taiwanensis and bacteria closely related to Paenibacillus stellifer, P. turicensis or Leptothrix sp. One mill was contaminated throughout with bacteria of a novel mesophilic genus most closely related to Brevibacillus centrosporus and another with bacteria of a novel thermophilic genus most closely related to Hydrogenophilus thermoluteolus. One B. cereus isolate producing haemolytic diarrhoeal enterotoxin was detected and all the tested B. licheniformis isolates produced a metabolite toxic to boar sperm cells., Conclusions: The bacilli and thermophilic bacteria isolated represent species which should not present occupational hazards in paper mill environments. The most harmful bacterium detected was B. licheniformis and potentially also B. cereus., Significance and Impact of the Study: Knowledge of the microbial diversity in a paper mill provides a rational basis for development of an effective controlling programme. A database constructed from the fingerprints generated using automated ribotyping helps to identify and trace the contamination routes of bacteria occurring in paper mills.

Formation of microbial biofilms and slimes is a general and serious problem in the operation of paper machines. Studies of microbial populations in paper machine-derived biofilms have been conducted using standard microbiological procedures; however, the bacterial genera present in this type of samples as well as their diversity are quite poorly known. Here, the bacterial diversity of 38 process water and 22 biofilm samples from four different Finnish paper machines were analyzed by length heterogeneity analysis of PCR-amplified 16S ribosomal DNA (LH-PCR). In addition, sequencing of the amplified 16S rRNA gene from 69 clones was conducted for characterization of the bacterial genera present in biofilm and slime samples. The LH-PCR profiles of both the free-living (process waters) and immobilized (biofilms) bacteria were diverse at all stages of the papermaking process. Out of the 69 sequenced clones, 44 belonged to alpha-Proteobacteria, most of which were close to the nitrogen-fixing root nodule genera Sinorhizobium, Rhizobium and Azorhizobium. Other clones were assigned to beta- and gamma-Proteobacteria and the phylum Bacteroidetes. In addition, eight of the clones were assigned to a yet uncultivated phylum, TM7. Finally, epifluorescence microscopy revealed that Gram-negative bacteria were predominant in both the biofilm (65%) and process water (54%) samples and a small coccoid cell morphology was most common in all samples. Together, our results show that the analysis of microbial samples from paper machines using modern molecular biology techniques adds valuable information and should, therefore, be useful as a more specific and sensitive microbiological method for the paper industry. This information could further be applied, e.g., in the development of more specific and environmental friendly antimicrobial agents for paper mills.

In the present study sequential anaerobic and aerobic treatment in two steps bioreactor was performed for removal of colour in the pulp and paper mill effluent. In anaerobic treatment, colour (70%), lignin (25%), COD (42%), AOX (15%) and phenol (39%) were reduced in 15 days. The anaerobically treated effluent was separately applied in bioreactor in presence of fungal strain, Paecilomyces sp., and bacterial strain, Microbrevis luteum. Data of study indicated reduction in colour (95%), AOX (67%), lignin (86%), COD (88%) and phenol (63%) by Paecilomyces sp. where as M. luteum showed removal in colour (76%), lignin (69%), COD (75%) AOX (82%) and phenol (93%) by day third when 7 days anaerobically treated effluent was further treated by aerobic microorganisms. Change in pH of the effluent, and increase in biomass of microorganisms substantiated results of the study, which was concomitant to the treatment method.

Over 100 bacteria from pulp and slime samples in a Canadian paper mill were identified by partial sequencing of their 16S rDNAs. Seventy-one percent of the isolates could be assigned to a bacterial genus with a high level of confidence. Another 12% exhibited at least 95% similarity within their 16S rDNA sequence with unidentified organisms that originate from warm or wet environments. Pseudomonas, Bacillus, and Pseudoxanthomonas isolates were represented at a relatively high proportion in both pulp and slime samples. This is the first time that Pseudoxanthomonas strains have been isolated from pulp and slime samples on a paper machine.

We investigated the in vitro influence of humic substances (HS) extracted from de-inking paper sludge compost on the inhibition of Pythium ultimum by two compost bacteria, Rhizobium radiobacter (Agrobacterium radiobacter) and Pseudomonas aeruginosa. When low concentrations (5 or 50 mg l(-1)) of HS were added to the culture medium, fungal inhibition by R. radiobacter significantly increased (P<0.01) by 2-3%. In contrast, these low levels of HS had no effect on P. ultimum inhibition by P. aeruginosa. The Fe, chelated by HS, was in part responsible for the decrease of P. ultimum inhibition by the bacteria when increasing amounts of HS were added in the culture medium. The addition of 500 mg l(-1) of humic acids isolated from de-inking paper sludge compost or from fossil origin completely eliminated the inhibition of P. ultimum by R. radiobacter. This Fe effect also stimulated growth of R. radiobacter and reduced its siderophore production in a minimal medium supplemented with HS as sole source of Fe. The results showed that HS influence microbial antagonism when added to a culture medium. However, this effect varies with different factors such as the type of bacteria, concentration of HS, molecular weight and Fe content.

To get insight into the microbial community of an Upflow Anaerobic Sludge Blanket reactor treating paper mill wastewater, conventional microbiological methods were combined with 16S rRNA gene analyses. Particular attention was paid to microorganisms able to degrade propionate or butyrate in the presence or absence of sulphate. Serial enrichment dilutions allowed estimating the number of microorganisms per ml sludge that could use butyrate with or without sulphate (10(5)), propionate without sulphate (10(6)), or propionate and sulphate (10(8)). Quantitative RNA dot-blot hybridisation indicated that Archaea were two-times more abundant in the microbial community of anaerobic sludge than Bacteria. The microbial community composition was further characterised by 16S rRNA-gene-targeted Denaturing Gradient Gel Electrophoresis (DGGE) fingerprinting, and via cloning and sequencing of dominant amplicons from the bacterial and archaeal patterns. Most of the nearly full length (approximately 1.45 kb) bacterial 16S rRNA gene sequences showed less than 97% similarity to sequences present in public databases, in contrast to the archaeal clones (approximately. 1.3 kb) that were highly similar to known sequences. While Methanosaeta was found as the most abundant genus, also Crenarchaeote-relatives were identified. The microbial community was relatively stable over a period of 3 years (samples taken in July 1999, May 2001, March 2002 and June 2002) as indicated by the high similarity index calculated from DGGE profiles (81.9+/-2.7% for Bacteria and 75.1+/-3.1% for Archaea). 16S rRNA gene sequence analysis indicated the presence of unknown and yet uncultured microorganisms, but also showed that known sulphate-reducing bacteria and syntrophic fatty acid-oxidising microorganisms dominated the enrichments.

Development of novel enzymatic methods for slime deposit control in paper mills requires knowledge of polysaccharide-producing organisms and the polysaccharide structures present in deposits. In this work, 27 polysaccharide-producing bacteria were isolated from slime samples collected from different parts of a paper machine. Most of the isolates produced polysaccharides in liquid culture and nine of them were selected for production of polysaccharides for characterisation. The selected isolates belonged to seven different genera: Bacillus, Brevundimonas, Cytophaga, Enterobacter, Klebsiella, Paenibacillus and Starkeya. Using ribotyping, partial 16S rDNA sequencing, physiological tests and fatty acid analysis, four of the nine isolates: Bacillus cereus, Brevundimonas vesicularis, K. pneumoniae and P. stellifer were identified to the species level. Production of polysaccharides by the selected isolates varied between 0.07 and 1.20 g L(-1), the highest amount being produced by B. vesicularis. The polysaccharides were heteropolysaccharides with varying proportions of galactose, glucose mannose, rhamnose fucose and uronic acids.

In this White Paper, we discuss the current state of microbial cancer therapy. This paper resulted from a meeting ('Microbial Based Cancer Therapy') at the US National Cancer Institute in the summer of 2017. Here, we define 'Microbial Therapy' to include both oncolytic viral therapy and bacterial anticancer therapy. Both of these fields exploit tumor-specific infectious microbes to treat cancer, have similar mechanisms of action, and are facing similar challenges to commercialization. We designed this paper to nucleate this growing field of microbial therapeutics and increase interactions between researchers in it and related fields. The authors of this paper include many primary researchers in this field. In this paper, we discuss the potential, status and opportunities for microbial therapy as well as strategies attempted to date and important questions that need to be addressed. The main areas that we think will have the greatest impact are immune stimulation, control of efficacy, control of delivery, and safety. There is much excitement about the potential of this field to treat currently intractable cancer. Much of the potential exists because these therapies utilize unique mechanisms of action, difficult to achieve with other biological or small molecule drugs. By better understanding and controlling these mechanisms, we will create new therapies that will become integral components of cancer care.

Thermophilic treatment systems are gaining more and more interest due to increasing system closure and higher process water temperatures. The use of activated sludge as a suitable post treatment system for anaerobically pretreated paper process was studied at 30 and 55 degrees C. Two lab-scale plug flow activated sludge reactors were run in parallel for 6 months at 30 and 55 degrees C. Both reactors were operated simultaneously at a 20, 15 and 10 days SRT. The effects of temperature and SRT on sludge settleability and COD removal efficiencies of different fractions were studied. Total COD removal percentages over the whole experimental period were 58 +/- 5% at 30 degrees C and 48 +/- 10% at 55 degrees C. The effect of the SRT on the total COD removal was negligible. Differences in total COD removal between both systems were due to a lesser removal of soluble and colloidal COD at 55 degrees C compared to the reference system. Two major characteristics of the thermophilic reactor were: 1. a lesser removal of soluble COD compared to the mesophilic reactor and 2. a turbid effluent compared to the mesophilic reactor. Thermophilic effluent turbidity was caused by a combination of influent colloidal particles that were not effectively retained in the sludge flocs and an erosion of thermophilic activated sludge itself, as shown by DGGE profiles. DGGE patterns of mesophilic sludge differed from the thermophilic sludge indicating that different microbial communities were present in both reactor systems.

The main objective of this study was to develop a system for the production of "renewable" hydrogen. Paper sludge is a solid industrial waste yielding mainly cellulose, which can be used, after hydrolysis, as a feedstock in anaerobic fermentation by (hyper)thermophilic organisms, such as Thermotoga elfii and Caldicellulosiruptor saccharolyticus. Tests on different medium compositions showed that both bacteria were able to produce hydrogen from paper sludge hydrolysate, but the amount of produced hydrogen and the requirement for other components differed. Hydrogen production by T. elfii strongly depended on the presence of yeast extract and salts. By contrast, C. saccharolyticus was less dependent on medium components but seemed to be inhibited by a component present in the sludge hydrolysate. Utilization of xylose was preferred over glucose by C. saccharolyticus.

One-dollar bills were collected from the general community in western Ohio to survey for bacterial contamination. Pathogenic or potentially pathogenic organisms were isolated from 94% of the bills. These results suggest a high rate of bacterial contamination of one-dollar bills.

Mechanisms for the undesired persistence of Bacillus species in paper machine slimes were investigated. Biofilm formation was measured for industrial Bacillus isolates under paper machine wet-end-simulating conditions (white water, pH 7, agitated at 45 degrees C for 1-2 days). None of the 40 tested strains of seven Bacillus species formed biofilm on polished stainless steel or on polystyrene surfaces as a monoculture. Under the same conditions, Deinococcus geothermalis E50051 covered all test surfaces as a patchy thick biofilm. The paper machine bacilli, however, formed mixed biofilms with D. geothermalis E50051 as revealed by confocal microscopy. Biofilm interactions between the bacilli and the deinococci varied from synergism to antagonism. Synergism in biofilm formation of D. geothermalis E50051 was strongest with Bacillus coagulans D50192, and with the type strains of B. coagulans, B. amyloliquefaciens or B. pumilus. Two B. licheniformis, one B. amyloliquefaciens, one B. pumilus and four B. cereus strains antagonized biofilm production by D. geothermalis. B. licheniformis D50141 and the type strain of B. licheniformis were the strongest antagonists. These bacteria inhibited deinococcal growth by emitting heat-stable, methanol-soluble metabolite(s). We conclude that the persistence of Bacillus species in paper machine slimes relates to their ability to conquer biofilms formed by primary colonizers, such as D. geothermalis.

Biofilm-degrading enzymes are potential agents for slime control in paper machines. In this work, extracellular polysaccharides were produced by bacteria isolated from paper machines and the isolated polysaccharides were used as substrates for the screening of polysaccharide-degrading microbes. Polysaccharide yields of 1.5-3.5 g/l were obtained by ethanol precipitation from cultures of strains of Klebsiella pneumoniae, Bacillus licheniformis and Pseudomonas fluorescens on sucrose medium. Two K. pneumoniae strains apparently produced an identical heteropolysaccharide containing galacturonic acid. Fructose-containing polysaccharides were the main products of B. licheniformis and P. fluorescens. Bacteria capable of hydrolyzing the fructose-containing polymers (levans) appeared to be relatively common among the strains selected for screening. None of the bacteria or mixed cultures screened were able to utilize the Klebsiella heteropolysaccharides.

The degradation of dissolved and colloidal substances from thermomechanical pulp (TMP) by bacteria isolated from a paper mill was studied in a laboratory slide culture system. Burkholderia cepacia strains hydrolysed triglycerides to free fatty acids, and the liberated unsaturated fatty acids were then degraded to some extent. Saturated fatty acids were not notably degraded. However, the branched anteiso-heptadecanoic fatty acid was degraded almost like the unsaturated fatty acids. About 30% of the steryl esters were degraded during 11 days, increasing the concentrations of free sterols. Approximately 25% of the dehydroabietic, and 45% of the abietic and isopimaric resin acids were degraded during 11 days. The degree of unsaturation seemed to be of greater importance for the degradation of fatty acids than the molar mass. No degradation of dissolved hemicelluloses could be observed with any of the nine bacterial strains studied. Burkholderia cepacia strains and one Bacillus coagulans strain degraded monomeric fructose and glucose in winter TMP water, but in summer TMP water, with much lower sugar concentrations, also other Bacillus strains degraded monomeric sugars.

The effect of pH, nutrient and aeration was studied on the removal of colour and reduction of BOD, COD and heavy metals with addition of readily available source of nitrogen and phosphorus in concentration of 1.0 g/L. Recalcitrant compound was effectively degraded by active microbial consortia. The isolated bacteria were identified as Pseudomonas putida (S1), Citrobacter sp. (S4) and Enterobacter sp. (S5). These organisms not only decolourised effluent upto 97% but reduced BOD, COD, phenolics and sulfide upto 96.63, 96.80, 96.92 and 96.67% respectively within 24 hrs of aeration and the heavy metals were removed upto 82-99.80%. The TSS and TDS were sharply reduced due to degradation. The absorption maxima was also decreased to 90%. However, in control without the microbial consortium no noticeable change was produced.

Due to the presence of antibiotics in environmental water and their potential influence on the occurrence of antibiotic-resistant bacteria, development of a detection method suitable for the screening of environmental water for antibiotics is required. In this study, we developed a simple colorimetric paper-based biosensor based on a novel principle for the detection of antibiotics inhibiting bacterial protein synthesis, including aminoglycosides, tetracycline, chloramphenicol, and macrolides. This biosensor is based on the detection of a color change induced by β-galactosidase, which is synthesized on freeze-dried paper discs containing an in vitro transcription/translation system. When a water sample without antibiotics is applied to the paper discs, β-galactosidase can be synthesized, and it hydrolyzes a colorimetric substrate, resulting in a color change from yellow to purple. By contrast, in the presence of antibiotics, the color change can be hampered due to an inhibition of β-galactosidase synthesis. We investigated the effect of the incubation temperature and pH of water samples and confirmed that the paper discs showed the color change to purple in the ranges of 15-37°C and pH 6-10. We observed concentration-dependent color variations of the paper discs by the naked eye and further estimated detection limits to be 0.5, 2.1, 0.8, and 6.1 μg/mL for paromomycin, tetracycline, chloramphenicol, and erythromycin, respectively, using digitized pictures. The paper-based biosensor proved to detect 0.5 μg/mL paromomycin, spiked in real environmental water samples, by the naked eye., (Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

The microbial content of printing paper machines, running at a temperature of 45-50 degrees C and at pH 4.5-5, was studied. Bacteria were prevalent colonizers of the machine wet end and the raw materials. A total of 390 strains of aerobic bacteria were isolated and 86% of these were identified to genus and species by biochemical, chemotaxonomic and phylogenetic methods. The most common bacteria found at the machine wet end were Bacillus coagulans and other Bacillus species, Burkholderia cepacia, Ralstonia pickettii, and in pink slimes, accumulating in the wire area and press section, species of Deinococcus, aureobacterium and Brevibacterium. Paper-making chemicals also contained species of Aureobacterium, B. cereus, B. licheniformis, B. sphaericus, Bordetella, Hydrogenophaga, Klebsiella pneumoniae, Pantoea agglomerans, Pseudomonas stutzeri, Staphylococcus and sometimes other enteric bacteria, but these did not colonize the process water. Yeasts and moulds were not present in significant numbers. A total of 131 strains were tested for their potential to degrade paper-making raw materials; 91 strains were found to have degradative activity, mainly species of Burkholderia and Ralstonia, Sphingomonas and Bacillus, and enterobacteria produced enzymes which degraded paper-making chemicals. Stainless steel adhering strains occurred in slimes and wire water and were identified as Burkholderia cepacia, B. coagulans and Deinococcus geothermalis. Coloured slimes were formed on the machine by species of Deinococcus, Acinetobacter and Methylobacterium (pink), Aureobacterium, Pantoea and Ralstonia (yellowish) and Microbulbifer-related strains (brown). The impact of the strains and species found in the printing paper machine community on the technical quality of paper, machine operation, and as a potential biohazard (Hazard Group 2 bacteria), is discussed.

Background: Antibiotic resistance (ABR) is one of the major health emergencies for global society. Little is known about the ABR of environmental bacteria and therefore it is important to understand ABR reservoirs in the environment and their potential impact on health., Method/design: Quantitative and qualitative data will be collected during a 3-year follow-up study of a river associated with religious mass-bathing in Central India. Surface-water and sediment samples will be collected from seven locations at regular intervals for 3 years during religious mass-bathing and in absence of it to monitor water-quality, antibiotic residues, resistant bacteria, antibiotic resistance genes and metals. Approval has been obtained from the Ethics Committee of R.D. Gardi Medical College, Ujjain, India (No. 2013/07/17-311)., Results: The results will address the issue of antibiotic residues and antibiotic resistance with a focus on a river environment in India within a typical socio-behavioural context of religious mass-bathing. It will enhance our understanding about the relationship between antibiotic residue levels, water-quality, heavy metals and antibiotic resistance patterns in Escherichia coli isolated from river-water and sediment, and seasonal differences that are associated with religious mass-bathing. We will also document, identify and clarify the genetic differences/similarities relating to phenotypic antibiotic resistance in bacteria in rivers during religious mass-bathing or during periods when there is no mass-bathing.

Background: The collection of subgingival plaque samples with paper points is time-consuming and accident-sensitive. However, the collection of saliva is simple and contains pathogens of all intraoral surfaces. The aim of this study is to investigate whether a sampling strategy with mouthrinse (mouthrinse sample [MSP]; test) leads to results comparable with standard sampling method (multiple site test from the deepest pocket of each quadrant [MT4]; control)., Methods: In 50 patients with periodontitis, subgingival plaque was sampled from the deepest pocket of each quadrant by using paper points and by gaining saliva with saline mouthrinse. Analysis was performed using a commercially available polymerase chain reaction test for 11 periodontal pathogens., Results: Detection frequency of Aggregatibacter actinomycetemcomitans (MT4/MSP: 42%/36%), Porphyromonas gingivalis (78%/66%), Tannerella forsythia (98%/84%), Treponema denticola (94%/74%), Parvimonas micra (86%/62%), Campylobacter rectus (90%/76%), Eubacterium nodatum (64%/30%), Prevotella intermedia (58%/54%), and Eikenella corrodens (90%/82%) was higher with MT4 than MSP. For Fusobacterium nucleatum (100%/100%), there was no difference between test and control. Only detection frequency of Capnocytophaga species (68%/74%) was higher with MSP than MT4. Differences were significant for P. gingivalis, T. forsythia, T. denticola, P. micra, C. rectus, and E. nodatum., Conclusions: There is no significant difference between MT4 and MSP for detection frequency of key pathogen A. actinomycetemcomitans. Key pathogens P. gingivalis, T. forsythia, T. denticola, P. micra, C. rectus, and E. nodatum show statistically higher detection frequencies with MT4.

In this Review, we explore natural product antibiotics that do more than simply inhibit an active site of an essential enzyme. We review these compounds to provide inspiration for the design of much-needed new antibacterial agents, and examine the complex mechanisms that have evolved to effectively target bacteria, including covalent binders, inhibitors of resistance, compounds that utilize self-promoted entry, those that evade resistance, prodrugs, target corrupters, inhibitors of 'undruggable' targets, compounds that form supramolecular complexes, and selective membrane-acting agents. These are exemplified by β-lactams that bind covalently to inhibit transpeptidases and β-lactamases, siderophore chimeras that hijack import mechanisms to smuggle antibiotics into the cell, compounds that are activated by bacterial enzymes to produce reactive molecules, and antibiotics such as aminoglycosides that corrupt, rather than merely inhibit, their targets. Some of these mechanisms are highly sophisticated, such as the preformed β-strands of darobactins that target the undruggable β-barrel chaperone BamA, or teixobactin, which binds to a precursor of peptidoglycan and then forms a supramolecular structure that damages the membrane, impeding the emergence of resistance. Many of the compounds exhibit more than one notable feature, such as resistance evasion and target corruption. Understanding the surprising complexity of the best antimicrobial compounds provides a roadmap for developing novel compounds to address the antimicrobial resistance crisis by mining for new natural products and inspiring us to design similarly sophisticated antibiotics., (© 2024. Springer Nature Limited.)

Gold Nanoparticles (AuNPs) have gained significant attention in biosensor development due to their unique physical, chemical, and optical properties. When incorporated into biosensors, AuNPs offer several advantages, including a high surface area-to-volume ratio, excellent biocompatibility, ease of functionalization, and tunable optical properties. These properties make them ideal for the detection of various biomolecules, including proteins, nucleic acids, and bacterial and viral biomarkers. Traditional methods for detecting bacteria and viruses, such as RT-PCR and ELISA, often suffer from complexities, time consumption, and labor intensiveness. Consequently, researchers are continuously exploring novel devices to address these limitations and effectively detect a diverse array of infectious pathogenic microorganisms. In light of these challenges, nanotechnology has been instrumental in refining the architecture and performance of biosensors. By leveraging advancements in nanomaterials and strategies of biosensor fabrication the sensitivity and specificity of biosensors can be enhanced, enabling more precise detection of pathogenic bacteria and viruses. This review explores the versatility of AuNPs in detecting a variety of biomolecules, including proteins, nucleic acids, and bacterial and viral biomarkers. Furthermore, it evaluates recent advancements in AuNPs-based biosensors for the detection of pathogens, utilizing techniques such as optical biosensors, lateral flow immunoassays, colorimetric immunosensors, electrochemical biosensors, and fluorescence nanobiosensors. Additionally, the study discusses the existing challenges in the field and proposes future directions to improve AuNPs-based biosensors, with a focus on enhancing sensitivity, selectivity, and their utility in clinical and diagnostic applications., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Methane is a potent greenhouse gas that enters the marine system in large quantities at seafloor methane seeps. At a newly discovered seep site off the coast of Point Dume, CA, ~ meter-scale carbonate chimneys host microbial communities that exhibit the highest methane-oxidizing potential recorded to date. Here, we provide a detailed assessment of chimney geobiology through correlative mineralogical, geochemical, and microbiological studies of seven chimney samples in order to clarify the longevity and heterogeneity of these highly productive systems. U-Th dating indicated that a methane-driven carbonate precipitating system at Point Dume has existed for ~20 Kyr, while millimeter-scale variations in carbon and calcium isotopic values, elemental abundances, and carbonate polymorphs revealed changes in carbon source, precipitation rates, and diagenetic processes throughout the chimneys' lifespan. Microbial community analyses revealed diverse modern communities with prominent anaerobic methanotrophs, sulfate-reducing bacteria, and Anaerolineaceae; communities were more similar within a given chimney wall transect than in similar horizons of distinct structures. The chimneys represent long-lived repositories of methane-oxidizing communities and provide a window into how carbon can be transformed, sequestered, and altered over millennia at the Point Dume methane seep., (© 2024 John Wiley & Sons Ltd.)

Previous studies have shown a significant amount of contaminants on paper currencies. It is important to study the transfer of microorganisms between paper currencies to determine whether it meets the level of a human health threat. This cross-contamination potential was analyzed by seeding new US 1-dollar bills with Bacillus thuringiensis, and pressing or rubbing them against clean currency to determine the amount of bacteria transfer to the unseeded currency. The transferred amount of bacteria was recovered, plated, incubated, and the colony-forming units were quantified. Among the recovery methods tested, the most efficient method, vortexing for 10 min with a recovery efficiency of 40 ± 8.1%, was used in this analysis. The resulting transfer rates were 4.8%, 8.6%, and 14.3% when pressed for 24 h, 72 h, and rubbed together, respectively. These transferred amounts of bacteria are significant and have the potential to spread infectious diseases., (Published 2016. This article is a U.S. Government work and is in the public domain in the USA.)

Bacteriological survey of one hundred twenty currency notes was done. Aerobic spore-forming bacilli (91%), Staphylococcus epidermidis (63.3%), Staphylococcus aureus (4.2%), Enterococcus (24.1%), alpha-hem. streptococcus (4.1%), Streptococcus pneumoniae (1.7%), Corynebacterium (7.5%), Lactobacilli (10.8%), Klebsiella pneumoniae (31.7%), Enterobacter (19.2%), E. coli (17.5%), Proteus (1.7%), Pseudomonas aeruginosa (0.8%), Shigella flexneri (0.8%) were isolated from paper money samples. Currency notes in general were bacteriologically contaminated especially with enteric pathogens and potentially pathogens, it was thought that some measures have to be taken to reduce these ill effects.

Pulp paper mill pollutants are the major source of aquatic contamination having metals, lignin and chlorophenols. Study was conducted to see the effect of these contaminants on antibiotic resistance pattern of isolated bacteria. Pulp paper effluents were evaluated for its physico-chemical properties i.e, BOD 72143 +/- 164.81 to 22.32 +/- 2.48, COD 213136 +/- 583.59 to 60.40 +/- 6.34, total phenol 386 +/- 71.24 to 0.43 +/- 0.0, lignin 26312 +/- 258.59 to 73.67 +/- 31.81and microbial quality i.e. K. pneumonae, S. typhi, S. faecalis, P. aeruginosa, E. coli, Ent. faecalis, A. hydrophila, B. subtilis, S. aureus, Y enterolitica and V vulrificus. Antibiotic sensitivity (10-30 microg), heavy metal resistance (100-1000 microg ml(-1)), lignin (1000-10,000 ppm) and pentachlorophenol (100-1000 ppm) tolerance of bacterial strains were assessed by seven classes of antibiotics. Eleven bacterial isolates were found multidrug resistant towards antibiotics, heavy metal, lignin and PCP. Out of 11 isolates, 90.9% were found resistant against eleven antibiotics which acquired 100% resistant in presence of heavy metal, lignin and chlorophenols. Results also revealed that concentration of lignin (50-350 ppm) and PCP (5-30 ppm) induced maximum growth (273-8050 cfu ml(-1)) of pathogenic bacteria in river water.

A rod-shaped, non-spore-forming, non-motile, aerobic, oxidase and catalase-positive and radiation-resistant bacterium (designated strain K4.1(T)) was isolated from biofilm collected from a Finnish paper mill. The bacterium grew as pale pink colonies on oligotrophic medium at 12 to 50 °C (optimum 37 to 45 °C) and at pH 6 to 10.3. The DNA G+C content of the strain was 66.8 l%. According to 16S rRNA gene sequence analysis, strain K4.1(T) was distantly related to the genus Deinococcus, sharing highest similarity with Deinococcus pimensis (90.0  %). In the phylogenetic tree, strain K4.1(T) formed a separate branch in the vicinity of the genus Deinococcus. The peptidoglycan type was A3β with L-Orn-Gly-Gly and the quinone system was determined to be MK-8. The polar lipid profile of strain K4.1(T) differed markedly from that of the genus Deinococcus. The predominant lipid of strain K4.1(T) was an unknown aminophospholipid and it did not contain the unknown phosphoglycolipid predominant in the polar lipid profiles of deinococci analysed to date. Two of the predominant fatty acids of the strain, 15 : 0 anteiso and 17 : 0 anteiso, were lacking or present in small amounts in species of the genus Deinococcus. Phylogenetic distinctness and significant differences in the polar lipid and fatty acid profiles suggest classification of strain K4.1(T) as a novel genus and species in the family Deinococcaceae, for which we propose the name Deinobacterium chartae gen. nov., sp. nov. The type strain is K4.1(T) (=DSM 21458(T) =HAMBI 2721(T)).

During the process of papermaking by pulp and paper plants, a thick and viscous deposits, termed slime, is quickly formed around the paper machines, which can affect the papermaking process. In this study, we explored the composition of the bacterial biota in slime that developed on shower pipes from 2 machines at a Canadian paper mill. Firstly, the composition was assessed for 12 months by DNA profiling with polymerase chain reaction coupled with denaturing gradient gel electrophoresis. Except for short periods (2-3 months), clustered analyses showed that the bacterial composition of the slime varied substantially over the year, with less than 50% similarity between the denaturing gradient gel electrophoresis profiles. Secondly, the screening of 16S rRNA gene libraries derived from 2 slime samples showed that the most abundant bacteria were related to 6 lineages, including Chloroflexi, candidate division OP10, Clostridiales, Bacillales, Burkholderiales, and the genus Deinococcus. Finally, the proportion of 8 bacterial lineages, such as Deinococcus sp., Meiothermus sp., and Chloroflexi, was determined by the Catalyzed Reporter Deposition-Fluorescence In Situ Hybridization in 2 slime samples. The results showed a high proportion of Chloroflexi, Tepidimonas spp., and Schlegelella spp. in the slime samples.

Bioelectrochemical reactors (BERs) with a cathodic working potential of -0.6 or -0.8 V more efficiently degraded cellulosic material, i.e., filter paper (57.4-74.1% in 3 days and 95.9-96.3% in 7 days) than did control reactors without giving exogenous potential (15.4% in 3 days and 64.2% in 7 days). At the same time, resultant conversions to methane and carbon dioxide in cathodic working chamber of BERs by application of electrochemical reduction in 3 days of operation were larger than control reactors. However, cumulative methane production in cathodic BERs was similar to those in control reactors after 7 days of operation. Microscopic observation and 16S rRNA gene analysis showed that microbial growth in the entire consortium was higher after 2 days of operation of cathodic BERs as compared with the control reactors. In addition, the number of methanogenic 16S rRNA gene copies in cathodic BERs was higher than in control reactors. Moreover, archaeal community structures constructed in cathodic BERs consisted of hydrogenotrophic methanogen-related organisms and differed from those in control reactors after 2 days of operation. Specifically, the amount of Methanothermobacter species in cathodic BERs was higher within archaeal communities than in those control reactors after 2 days of operation. Electrochemical reduction may be effective for accelerating microbial growth in the start-up period and thereby increasing microbial treatment of cellulosic waste and methane production.

The main objective of the present study was to improve the quality of pulp and paper industrial wastewater of two local mills RAKTA and El-Ahlia, Alexandria, Egypt, and to bring their pollutant contents to safe discharge levels. Quality improvement was carried out using integrated chemical and biological treatment approaches after their optimization. Chemical treatment (alum, lime, and ferric chloride) was followed by oxidation using hydrogen peroxide and finally biological treatment using activated sludge (90 min for RAKTA and 60 min for El-Ahlia effluents). Chemical coagulation produced low-quality effluents, while pH adjustment during coagulation treatment did not enhance the quality of the effluents. Maximum removal of the tested pollutants was achieved using the integrated treatment and the pollutants recorded residual concentrations (RCs) of 34.67, 17.33, 0.13, and 0.43 mg/l and 15.0, 11.0, 0.0, and 0.13 mg/l for chemical oxygen demand (COD), biochemical oxygen demand (BOD5), tannin and lignin, and silica in RAKTA and El-Ahlia effluents, respectively, all of which were below their maximum permissible limits (MPLs) for the safe discharge into water courses. Specific oxygen uptake rate (SOUR) and sludge volume index (SVI) values reflect good conditions and healthy activated sludge. Based on the previous results, optimized conditions were applied as bench scale on the raw effluents of RAKTA and El-Ahlia via the batch chemical and the biological treatment sequences proposed. For RAKTA effluents, the sequence was as follows: (1) coagulation with 375 mg/l FeCl3, (2) oxidation with 50 mg/l hydrogen peroxide, and (3) biological treatment using activated sludge with 2,000 mg/l initial concentration and 90 min hydraulic retention time (HRT), while for El-Ahlia raw effluents, the sequence was (1) coagulation with 250 mg/l FeCl3, (2) oxidation with 45 mg/l hydrogen peroxide, and (3) biological treatment using activated sludge with 2,000 mg/l initial concentration and 60 min HRT. In conclusion, results confirmed that the application of the proposed sequential treatments removed almost all COD, BOD5, high molecular weight compounds, and silica from RAKTA and El-Ahlia influents and produced high-quality effluents, thus achieving the main objective of this study.

A process for production of polyhydroxyalkanoates (PHA) by activated sludge treating a paper mill wastewater was investigated. The applied strategy was to select for glycogen accumulating organisms (GAOs) by alternating anaerobic/aerobic conditions. Acidogenic fermentation was used as pretreatment to convert various organic compounds to volatile fatty acids which are preferable substrates for PHA production. Enrichment resulted in a culture dominated by GAOs related to Defluviicoccus vanus (56%) and Candidatus Competibacter phosphatis (22%). Optimization of PHA accumulation by the enriched GAO culture was performed through batch experiments. Accumulation of PHA under anaerobic conditions was limited by the intracellular glycogen stored. Under aerobic conditions significant glycogen production (to 25% of sludge dry weight) was observed alongside PHA accumulation (to 22% of sludge dry weight). By applying a subsequent anaerobic period after an initial aerobic, the produced glycogen could be utilized for further PHA accumulation and by this strategy PHA content was increased to 42% of sludge dry weight. The PHA yield over the entire process was 0.10 kg per kg of influent COD treated which is similar to what has been achieved with a process applying feast/famine enrichment strategy with the same wastewater., (IWA Publishing 2008.)

The metagenomic DNA of pulp sediments from paper mill effluent was extracted and purified. The 16S rDNA was amplified using the purified metagenomic DNA as template and a 16S rDNA library was prepared. Sequence analysis of 16S rDNA clones showed that diverse of uncultured bacteria inhabit in this environment, which can be classified into 4 clusters as Spirochaetes, Proteobacteria, Bacteroidetes and Firmicutes. A metagenomic library containing 10000 clones was constructed into cosmid vector, and the capacity of inserted DNA of which was 3.53 x 10(8) bp. Functional screening of the library resulted in isolation of two independent clones expressing endoglucanase activity, three independent clones expressing exoglucanase activity and two independent clones expressing beta-glucosidase activity. One clone expressing strongest enzyme activity from each activity category was chosen to be further analyzed. Three novel cellulase genes designated as umcel5L, umcel5M and umbgl3D were identified by subcloning, sequencing and expression. The umcel5L encodes an endoglucanase belonging to glycosyl hydrolase family 5, which is most related to an endoglucanase from Bradyrhizobium japonicum at 43% identity and 59% similarity. The umcel5M encodes a cellodextrinase belonging to glycosyl hydrolase family 5, which is most similar to a cellodextrinase from Fibrobacter succinogenes at 48% identity and 69% similarity. The umbgl3D encodes a putative beta-glucosidase belonging to glycosyl hydrolase family 3, which shares highest homology with a beta-glucosidase from Thermotoga maritima at 46% identity and 61% similarity. It is the first time to reveal the bacterial diversity of pulp sediments from paper mill effluent and clone novel cellulase genes from the bacteria by culture-independent method.

Knowledge of the microbiology of dinitrogen (N2)-fixing bacteria in compost rich in de-inking paper sludge (DPS) is limited. Dinitrogen (N2)-fixing bacteria from DPS composts were isolated and studied for their N2-fixing activity in vitro and in vivo. Two Gram-negative N2-fixing isolates were identified as Pseudomonas. At 20 degrees C, both isolates revealed that N2-fixing activity was higher than that of three arctic Pseudomonas strains. Their N2-fixing activity was found to occur between 18 and 25 degrees C, a pattern that was similar to the reference isolate Azotobacter ATCC 7486. Composts successfully showed N2-fixing activity after carbohydrate amendments both with and without inoculation of a N2-fixing isolate. These results suggest that DPS composts support N2-fixing bacteria and that N2-fixing activity is dependent on a usable carbohydrate source.