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Advances in liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) have enabled the quantification of immunosuppressants using microsampling techniques. In this context, dried matrix on paper discs (DMPD) could be a useful alternative to conventional venipuncture. Although analytical validation is necessary to establish the suitability of method performance, it is not sufficient to proceed with its implementation into routine clinical practice. Also necessary is that equivalence between sampling methods be demonstrated in a clinical validation study.To clinically validate a LC-MS/MS method for the quantification of tacrolimus, sirolimus, everolimus and cyclosporin A using DMPD.According to the recommendations of international guidelines, at least 40 whole blood (WB) and DMPD paired samples for each analyte were collected by skilled technicians and analyzed using LC-MS/MS. Results were evaluated in terms of statistical agreement and bias values at medical decision points.For all analytes, Passing-Bablok regression analysis revealed that confidence intervals (CIs) for slopes and intercepts included 1 and 0, respectively. It also showed that biases at medical decision points were not clinically relevant. No statistically significant differences between DMPD and WB were found using difference plots and agreement analysis. In this regard, CIs for bias estimators included 0, and more than 95% of the results fell within the limits of agreement.The feasibility of the clinical application of simultaneous quantification of tacrolimus, sirolimus, everolimus and cyclosporin A in DMPD was demonstrated. Results showed that this microsampling technique is interchangeable with conventional WB sampling when specimens are collected by trained personnel.

Research on the order Saprolegniales (Oomycota) has been an ongoing quest for more than a century. The best studied genera are Saprolegnia and Aphanomyces, known for their pathogenicity on freshwater animals. In this study, we reviewed 1073 papers and 2803 ITS sequences of Saprolegniales to investigate their taxonomy, diversity and potential roles in mainly freshwater ecosytems. We found that, in general, our knowledge on diversity and ecology of Saprolegniales is limited. Neither classic taxonomy nor available molecular techniques have been sufficient to delineate genera and species and show their relative distribution in freshwater-associated habitats. Also, we currently lack a comprehensive understanding of their involvement in carbon turnover and food web dynamics. Finally, due to lack of using high-throughput sequencing techniques, it is not clear how and to what extent communities of Saprolegniales might differ in freshwater econiches. Therefore, we provide a historical perspective on the establishment of Saprolegniales, explain improvements, highlight deficiencies, and finally propose new research avenues for more systematic studies. We conclude that challenges in studying Saprolegniales can be removed by increasing the practicality of classic taxonomy and applying available molecular toolboxes (multi-gene phylogeny and high-throughput sequencing). Additionally, inclusion of Saprolegniales in freshwater carbon cycling should be addressed for their better ecological resolution.

• The disinfection of mouldy paper-based objects is necessary because it ceases biodeterioration and eliminates viable microorganisms.

A bacterium isolated from effluent of thermal paper industry and identified as Pseudomonas aeruginosa (PAb1) based on 16SrRNA gene sequence analysis which could grow on basal mineral salt medium upon bisphenol A, which functions as an exclusive carbon source. Physicochemical variables of thermal paper industry effluent noted were significantly greater than the typical limit due to pollution of the acquiring water systems. The mathematic kinetic models like Monod, Moser and Tesier models were applied for batch fermentation of bisphenol A degradation in basal salt medium and the half saturation coefficient (KS) and the regression coefficient R2 using Monad, Moser and Tesier kinetic models registered as 9.947 g/L, 12.46 g/L and 14.14 g/L and 0.91, 0.94 and 0.84 respectively. Besides, the utmost specific growth rate μmax was witnessed as 0.841 h−1 for the P. aeruginosa (PAb1) regarding BPA degradation. Metabolic intermediates like phenol, acetophenone, and hydroquinone and p-hydroxybenzoic acid were also determined through the degradation process by GC-MS. The metabolic pathway of BPA degradation by the bacterial isolates was also designed in today's analysis. A probabilistic statistical model originated using Box-Behnken response surface methodology and process variables were optimized by nonlinear optimization.

The application of anaerobic digestion is mounting worldwide because of its economic and environmental benefits. As a concern, the rate and extent of the anaerobic digestion of pulp and paper mill sludge was tested using the biochemical methane potential (BMP) with different food/microorganisms (F/M) ratio. The study was evaluated at five different F/M ratios (i.e. 0.5, 1.0, 1.5, 2.0, 2.5). In addition to that reaction kinetics and the influence of biodegradability were also evaluated for the biomethane potential of substrates. The highest cumulative methane yield was obtained in F/M ratio 2.0 with 3.4 L followed by F/M ratio 1.5 and 2.5 with 3.3 L and 2.9 L, respectively. The maximum methane production rate 3.66 L CH4 was perceived as per kinetics. The biomass activity may be affected due to the high substrate concentration during BMP test of lower F/M ratios. On the other hand, F/M ratio greater than 2.0, methane yield was decreased due to increased volatile fatty acid production reduces the reactor pH and inhibits methanogens activity.

In the past, the animal glues were used in paper manufacturing and in restoration of artworks preserved in museums, libraries and archives. By ageing they went through deterioration creating distortions, tensions, cockling and discolouration in paper materials. Consequently, the removal of glue residues becomes an essential step in the restoration of ancient and artwork paper material. Current mechanical and chemical methods display serious drawbacks mainly related to aggressiveness towards material or toxicity for restorers. Bio-based methods for paper cleaning rely on the use of enzymes that require skilled operators, optimal application conditions and high costs, creating difficulties in mastering enzyme use so far. This paper describes a first attempt of biocleaning ancient paper from organic deposits using living bacteria. The non-pathogenic, non-spore-forming and non-cellulolytic original strain Ochrobactrum sp. TNS15 E was successfully applied -immobilised in an agar gel-on original paper specimens dating back to the 17th. After 4 h of contact with the bacterial pack, the cellulose fibres underlying glue were disclosed, highlighting the bacterial capacity of removing the glue layer without damaging the paper or leaving undesirable residues. Both colorimetry and SEM analyses proved the results. The procedure is simple, low-cost and safe for the artefact, the restorers and the environment.

Neisseria meningitidis (N. meningitidis), Streptococcus pneumoniae (S. pneumoniae), and Haemophilus influenzae type b (Hib) are three most common pathogens accounting for most bacterial meningitis, a serious global infectious disease with high fatality, especially in developing nations. Because the treatment and antibiotics differ among each type, the identification of the exact bacteria causing the disease is vital. Herein, we report a polymer/paper hybrid microfluidic biochip integrated with loop-mediated isothermal amplification (LAMP) for multiplexed instrument-free diagnosis of these three major types of bacterial meningitis, with high sensitivity and specificity. Results can be visually observed by the naked eye or imaged by a smartphone camera under a portable UV light source. Without using any specialized laboratory instrument, the limits of detection of a few DNA copies per LAMP zone for N. meningitidis, S. pneumoniae and Hib were achieved within 1 hour. In addition, these three types of microorganisms spiked in artificial cerebrospinal fluid (ACSF) were directly detected simultaneously, avoiding cumbersome sample preparation procedures in conventional methods. Compared with the paper-free non-hybrid microfluidic biochip over a period of three months, the hybrid microfluidic biochip was found to have a much longer shelf life. Hence, this rapid, instrument-free and highly sensitive microfluidic approach has great potential for point-of-care (POC) diagnosis of multiple infectious diseases simultaneously, especially in resource-limited settings.

In the present study, a phenol degrading actinomycetes strain was isolated from paper pulp wastewater. The isolated strain was identified as Rhodococcus pyridinivorans PDB9TN-1 using 16S rRNA gene sequencing technique, which yielded the complete degradation of 1600 mg l−1 of phenol within 56 h. Moreover, the Rhodococcus species could remove over 97.5% of total organic carbon (TOC) at an initial phenol concentration of 1000 mg l−1. The metabolic profile of phenol biodegradation by the actinomycetes species has been investigated using liquid chromatography–mass spectrometry (LC-MS) and Fourier-transform infrared spectroscopy (FT-IR). The key intermediate products detected in the actinomycetes culture were hydroquinone, protocatechuic acid, beta-carboxycis-cismuconate, maleyl acetate, 4–oxalocrotonic acid and 3-fumarylpyruvic acid. Further, a tentative oxalocrotonic acid pathway has been proposed on the basis of the detected intermediate products. The culture medium constituents are highly indispensable for the microbial growth, enzyme and other cellular metabolic activities associated with phenol biodegradation. Therefore, the effect of media components were modelled with the help of response surface methodology (RSM) and central composite design (CCD). A reliable quadratic model was established which predicted the degradation efficiency as high as 99.5% with an appreciable correlation coefficient (R2) of 0.98. Moreover, phenol degradation efficiency was improved with concentrations beyond the central values for all media components except K2HPO4 and NaCl.

Potential high bio-deterioration of certain microorganisms due to their metabolic activities on organic materials causes serious problems in the conservation of cultural heritage. This study focuses on establishing an efficient conservation strategy against microbial deterioration using biosynthesized nanoparticle (NPs). Isolation, identification, and characterization of microorganisms involved in deterioration of archaeological manuscript dating back to 17th century (1677 A.D) were conducted. Eleven bacterial species and fifteen fungal species were obtained and identified using 16S rRNA and ITS sequences, respectively. All strains exhibited potent cellulolytic activity. Bacillus subtilis strain (B3) and Penicillium chrysogenum strain (F9) achieved the highest cellulolytic activity amongst obtained bacterial and fungal strains. Paper models conservation effect of biosynthesized silver and zinc oxide NPs against strain (B3) and strain (F9) was evaluated. Microbial growth inhibition, color change according to CIE L*a*b* color space, fourier transform infrared spectroscopy (FT-IR) were assessed. Furthermore, morphology and the elemental component of paper models were analyzed using scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX). Silver NPs achieved better results than zinc oxide NPs for paper preservation. Application of 1.0 or 2.0 mM silver NPs exhibited the best preservation effect on the paper models achieving 100% microbial inhibition (bacteria and fungi, respectively). Moreover, NPs-treated paper models showed slight color change and exhibited similar structural analysis as original papers. These results showed the potentiality of this green strategy to be applied for conservation of cellulose-constructed cultural heritage.

Pulp and paper industry primary sludge being largely composed of lignocellulosic fibres, it could be used as carbon source by bacteria having cellulolytic capability. The aim of this study was to evaluate the use of cellulose contained in this type of sludge for Clostridium thermocellum to produce ethanol, hydrogen and cellulases. In an ATCC 1191 medium containing 5 kg m−3 dry primary sludge from recycled paper mill, batch culture reached stationary phase after 2 days. All of the available cellulose was hydrolysed after 60 h of incubation, with a final pH of 5.83. Metabolites produced after 60 h of fermentation were acetate (8.50 mol m−3), ethanol (11.30 mol m−3), lactate (8.75 mol m−3), formate (0.27 mol m−3), hydrogen (11.20 mol m−3) and carbon dioxide (18.41 mol m−3). Cellulase activity was detected in the supernatant after 36 h, with a maximal activity of 0.25 U cm−3 at 72 h. Pulp and paper primary sludge appeared to be a readily usable substrate for C. thermocellum at this concentration, yielding both potential biofuels (hydrogen and ethanol) as well as active cellulases.

Food-borne pathogens have been recognized as a major cause of human infections worldwide. Their identification needs to be simpler, cheaper and more reliable than the traditional methods. Here, we constructed a low-cost paper platform for viable pathogenic bacteria detection with the naked eye. In this study, an effective isothermal amplification method was used to amplify the hlyA mRNA gene, a specific RNA marker in Listeria monocytogenes. The amplification products were applied to the paper-based platform to perform a visual test using sandwich hybridization assays. When the RNA products migrated along the platform by capillary action, the gold nanoparticles accumulated at the designated area. Under optimized experimental conditions, as little as 0.5 pg/μL genomic RNA from L. monocytogenes could be detected. It could also be used to specifically detect 20 CFU/mL L. monocytogenes from actual samples. The whole assay process, including RNA extraction, amplification, and visualization, can be completed within several hours. This method is suitable for point-of-care applications to detect food-borne pathogens, as it can overcome the false-positive results caused by amplifying nonviable L. monocytogenes. Furthermore, the results can be imaged and transformed into a two-dimensional bar code through an Android-based smart phone for further analysis or in-field food safety tracking.

The present study deals with the bioremediation of potentially hazardous pulp and paper mill effluent by a laccase producing Paenibacillus sp. strain LD-1 (JX499920) isolated from contaminated soil sample by lignin enrichment method. The bacterium effectively reduced pollution parameters (colour 68%, lignin 54%, phenol 86%, BOD 83% and COD 78%) after 144 h of treatment at 34 ± 1 °C and 120 rpm. GC–MS analysis of control and treated samples showed that concentration of most of the low molecular weight phenolic compounds like 2-methoxyphenol, 2,6-dimethoxy phenol, 2-methoxy-4-ethyl-phenol, 3-allyl-6-methoxyphenol, ethanone 1-(-4-hydroxy-3,5-dimethoxyphenyl), benzoic acid, 2-methoxy-4-(1-propenyl) phenol and 4-methoxycinnamic acid present in control untreated effluent were reduced after bacterial treatment. The toxicity assessments were carried out with treated and untreated effluent by studying the growth and germination of seeds of mung bean (Vigna radiata L.). The mung bean bioassay confirmed the detoxification of effluent after bacterial treatment.

We investigated whether dried cerebrospinal fluid (CSF) conserved on filter paper can be used as a substrate for accurate PCR diagnosis of important causes of bacterial meningitis in the Lao PDR. Using mock CSF, we investigated and optimized filter paper varieties, paper punch sizes, elution volumes and quantities of DNA template to achieve sensitive and reliable detection of bacterial DNA from filter paper specimens. FTA Elute Micro Card™ (Whatman, Maidstone, UK) was the most sensitive, consistent and practical variety of filter paper. Following optimization, the lower limit of detection for Streptococcus pneumoniae from dried mock CSF spots was 14 genomic equivalents (GE)/μL (interquartile range 5.5 GE/μL) or 230 (IQR 65) colony forming units/mL. A prospective clinical evaluation for S. pneumoniae, S. suis and Neisseria meningitidis was performed. Culture and PCR performed on fresh liquid CSF from patients admitted with a clinical diagnosis of meningitis (n = 73) were compared with results derived from dried CSF spots. Four of five fresh PCR-positive CSF samples also tested PCR positive from dried CSF spots, with one patient under the limit of detection. In a retrospective study of S. pneumoniae samples (n = 20), the median (IQR; range) CSF S. pneumoniae bacterial load was 1.1 × 104 GE/μL (1.2 × 105; 1 to 6.1 × 106 DNA GE/μL). Utilizing the optimized methodology, we estimate an extrapolated sensitivity of 90%, based on the range of CSF genome counts found in Laos. Dried CSF filter paper spots could potentially help us to better understand the epidemiology of bacterial meningitis in resource-poor settings and guide empirical treatments and vaccination policies. © 2013 The Authors. Clinical Microbiology and Infection published by John Wiley and Sons Ltd on behalf of the European Society of Clinical Microbiology and Infectious Disease.

The aim of this study was to determine the cause of foxing on 19th century paper using omics methods, including metagenomics via high-throughput sequencing on the Illumina platform and metabolomics via high-resolution surface-assisted laser desorption/ionization time-of-flight mass spectrometry (SALDI-ToF-MS) imaging using the gold nanoparticle-enhanced target (AuNPET) method. Metabolomic analysis as well as ninhydrin and amido black staining of paper samples showed the presence of proteinaceous substances in areas corresponding to foxing. Products of cellulose biodegradation and microbial metabolites, including yellow-coloured pigments (2-methyl-6-phytylquinol; delta-, gamma- and beta-tocopherols; 3-hydroxy-L-kynurenine) were detected. No difference in metal-ion content was seen between areas with and without foxing. Higher degrees of biodiversity were identified within foxing stains compared to foxing-free areas. Phoma sp. and Cladonia sp. moulds and Gluconobacter and Ralstonia bacteria were predominant in DNA samples obtained from foxing stains. The presence of these microbes has never previously been linked to the phenomenon of foxing. These studies indicate that foxing is a result of a combined effect of microbial action, paper-degradation by microbes, dye production, and chemical changes occurring in paper due to oxidation of cellulose metabolites.

The main objective of this paper is the optimization and enhancing of the preservation quality of the carboxymethyl cellulose (CMC) polymer against microorganism factors by using titanium dioxide and TiO2/chitosan nanoparticles. Biological and thermal accelerated aging experiments were carried out on adhesive films and treated paper samples to investigate the influence of TiO2 and TiO2/chitosan nanoparticles on the properties of the aged adhesive films and treated papers. The antifungal activity of TiO2 and TiO2/chitosan nanoparticles was investigated against the two fungi species commonly found on library materials: Aspergillus flavus and Aspergillus niger. Our results showed that TiO2 is a desirable material for preserving CMC films and its treated papers, but its problem is that its restricted reactivity in the UV wavelength limits its use in indoor circumstances. TiO2/chitosan nanoparticles are more effective, even in darkness, and could be successfully applied to optimize the CMC properties. Furthermore, results show that the TiO2/chitosan nanoparticles decrease the yellowness of CMC adhesive and while not reducing its adherence. Also, this nanoparticle can obviously reduce the oxidation of paper during the aging process, and a slight decrease in the tensile strength and pH of the paper is observed.

A sample set of model graded papers containing known organic and inorganic components was used to culture an Aspergillus terreus strain able to produce acidic metabolites and cellulolytic enzymes. This study set out to document the biogenic formation of calcium oxalate crystals, as a consequence of fungal activity and growth, on a range of paper samples. Scanning electron microscopy and FTIR spectroscopy techniques were employed to achieve this end. Bioleaching processes induced by fungi on carbonate used in the manufacture of paper, together with the capacity of fungi to produce - very rapidly - significant biogenesis of monohydrate and dihydrate calcium oxalate crystals on the surface of papers, were investigated. SEM images of calcium containing paper samples disclosed a very clear difference in the pattern of inorganic material distribution when areas spoiled by fungi and those not affected by fungal growth were compared. In addition, the FTIR spectrum of CaCO(3) sized paper clearly demonstrated, following fungal growth, the FTIR absorbance of fungus and calcium oxalate, together with the disappearance of carbonate bands. (C) 2010 Elsevier Ltd. All rights reserved.

Since its invention, paper has become one of the main carriers of our cultural, scientific, political, economic and historical information. Given the importance of this material, its preservation is a matter of great interest. Paper can be deteriorated due to physical, chemical and biological agents. Within microorganisms, fungi are the major paper biodeteriogens. Throughout history, several methods have been used to prevent and stop fungal deterioration on paper based materials. In this work we present a review of the main chemical and physical methods used to avoid fungal paper biodeterioration until nowadays and also of some new approaches tested recently. The advantages and disadvantages of these methods are discussed as well as their health effects. Studies regarding antifungal compositions, methods of application, performance and effects on the treated materials are also presented with the aim of providing a clear set of conclusions on the topic.

Summary Background Infection with Staphylococcus aureus was initially considered a major problem in hospitals, but over the last few decades the incidence of community-acquired infection has also increased. Paper currency has recently been identified as another mode of spread by which community-acquired S. aureus infection may be transmitted, since paper currency is frequently transferred from one person to another. In the present study, S. aureus strains were isolated from paper currency and screened to detect virulence-associated genes. Methods Paper currency was collected randomly from operators in mutton shops, vegetable shops, hospitals, medical stores, snacks corners, and restaurants in the two cities of India, Dehradun and Delhi. Samples were screened for pathogenic S. aureus by various biochemical assays. Three isolates were used to study the survival of S. aureus on paper currency. Virulence genes were amplified by PCR. Antibiotic susceptibility was determined against nine antibiotics by the disk diffusion method. Results Sixty-seven isolates of Staphylococcus aureus were isolated from paper currency from the two Indian cities. The maximum number of isolates was recovered from hospitals, followed by mutton shops, and the minimum from restaurants. The test isolates could survive on paper currency for eight days under in vitro conditions (27°C temperature and 30% relative humidity). A set of four virulence genes viz. cna (16 isolates), icaA (19 isolates), hlg (21 isolates), and sdrE (18 isolates) was amplified using gene-specific primers. Only eight isolates possessed all four virulence genes. Antimicrobial susceptibility tests showed that those isolates having all the tested virulence genes were more resistant. Conclusions This study clearly suggests that paper currency can serve as a carrier for the spread of resistant bacterial pathogens.

Foxing of paper is a deterioration phenomenon occurring in the form of brown-yellowish spots, the abiotic and/or biotic causes of which are not yet completely understood. Nevertheless, microbiological infection has been recognized that may contribute to paper damage and therefore it becomes important to know the taxonomic position and the degradative activity of the potential infectious biological agents which mostly are fungi, but also bacteria and yeasts. A cellulolytic bacterial strain isolated from a foxed paper sample exhibited morphological and physiological characteristics of the Bacillus genus. To study its taxonomic position, different identification methods were used: the Biolog system, the direct amplified polymorphic DNA-polymerase chain reaction analysis (DAPD-PCR) and the partial sequencing of the 16S rDNA gene. Biolog system and partial sequencing of 16S rDNA gene assigned the strain to the Paenibacillus polymyxa species. DAPD-PCR analysis indicated a high similarity with Bacillus circulans, by comparing the isolated strain with some closely related Bacillus species.

A deacidifying/antifungal mixture composed of calcium propionate, methylparaben and propylparaben was tested against Aspergillus niger, Cladosporium cladosporioides, Chaetomium globosum, Penicillium chrysogenum and Penicillium corylophilum. The preventive treatment of paper samples resulted in a complete fungal growth inhibition on 4 of the 5 tested species. The antifungal properties of the formulation remained unaffected for a minimum period of one year. The disinfecting treatment with the mixture led to a total elimination of all tested fungal species. The effects of the tested formulation on paper were evaluated in terms of pH, colourimetry, folding endurance, FTIR and XRD, using moist heat artificial ageing. Aside from plain paper, paper previously colonized by A. niger was tested to evaluate the potential of the formulation in preventing deterioration caused by fungal metabolites. In plain paper, an effective deacidification and long-term prevention of mechanical resistance loss were achieved, although a slight paper discoloration occurred. On previously colonized by fungi, the treatment effectively prevented the deterioration caused by fungal metabolites.

Eight aerobic bacterial strains were isolated from pulp paper mill effluent sludge. Out of eight through nutrient enrichment technique three potential aerobic bacterial strains ITRC S(6), ITRC S(7) and ITRC S(8) were found capable to effectively degrade the kraft lignin (KL), a major byproduct of the chemical pulping process and main contributor to the colour and toxicity of effluent. Further, these potential strains (ITRC S(6), ITRC S(7) and ITRC S(8)) were biochemically characterised as Gram variable small rod, Gram negative rod and Gram positive rod respectively. Subsequently, 16S rRNA sequencing showed 95% base sequence homology and it was identified as Paenibacillus sp. (AY952466), Aneurinibacillus aneurinilyticus (AY856831), Bacillus sp. (AY952465) for ITRC S(6), IITRC S(7) and ITRC S(8), respectively. In batch decolourization experiments Bacillus sp. ITRC S(8) reduced the colour of lignin amended mineral salt medium, pH 7.6 by 65% after 6th d, at 30 degrees C, A. aneurinilyticus ITRC S(7) by 56% and Paenibacillus ITRC S(6) 43%. Under these conditions the three strains degraded the KL by 37%, 33% and 30%, respectively while the mixed culture of these three bacteria reduced colour by 69%, lignin by 40% and total substrate by 50% under same conditions. Biodegradation of the KL was not affected by low (

A pure culture of 2,4-dichlorophenol (2,4-DCP)-degrading bacteria was isolated from a natural enrichment that had been adapted to chlorophenols in the aeration pond of the Baikalsk pulp and paper mill (Russia). The bacteria were identified by 16S rDNA intergenic region analysis, using PCR with universal primers. Comparative analysis of the 16S rDNA sequence (1545 bp) in the GenBank database revealed that these bacteria are related to Bacillus cereus GN1. Degradation of 2,4-DCP was studied using this culture in liquid medium under aerobic conditions, at initial concentrations of 20–560mM 2,4-DCP. The 2,4-DCP degradation rates by B. cereus GN1 could be determined at concentrations up to 400mM. However, higher concentrations of 2,4-DCP (560mM) were inhibitory to cell growth. r 2006 Elsevier Ltd. All rights reserved.

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.

Extracts of naturally and artificially blotched postage stamps reacted positively to phenolreagents in three different tests. Microscopic examination of blotches revealed sparse living and dead hyaline hyphae. Six species of Aspergillus and one each of Penicillium and Gliocladium, isolated from foxed stamps, were capable of producing pigments in agar media and filter paper, sometimes more intense when grown in pairs than alone. All fungal isolates showed some degree of cellulolytic ability; all but A. fumigatus and G. roseum were capable of germinating, growing, sporulating, and producing pigments at high osmotic pressures, and all were capable at low osmotic pressure. Of three species inoculated on new stamp paper, one, A. terreus var. aureus, developed hyphae and fluorescent yellow and orange-brown pigments and produced pigments at constant relative humidity of 32.5% over an eight month period. Ferrous and ferric salts appeared to be unnecessary for production of pigments diffused into agar media, liquid medium, or filter paper, but in some isolates they intensified the color. Stamps inoculated with A. terreus var aureus conidia and irradiated with a dose of 2.5 Mrad remained free of blotches throughout the two year experimental period.

Fungal stains affect documents and artworks on paper all over the world, diminishing their chemical stability and compromising their readability. The present paper studies the suitability of agarose and gellan gum hydrogels to remove fungal stains from paper, using paper impregnated with alizarin as a model system to simulate the most common colorant molecules produced by fungi - polyketide quinones. The effect of pH variation on the efficacy of the gels was evaluated by UV spectrometry. The results show that the cleaning efficacy of the gels greatly depends on the gel matrix, the colorant molecules, and the pH balance of the process.

The effect of pH adjustment and surfactant addition on the anaerobic degradation of sludge from a pulp and paper mill waste treatment plant was monitored by COD (Chemical Oxygen Demand) and EOX (Extractable Organic Halogen) analysis. COD decreased by 29–64% and EOX by 22–50%, in all treatments (including control, non-adjusted, sludges). Adjusting the initial pH to 7.0 with sodium bicarbonate, but not with potassium hydroxide, accelerated degradation. Maximum decreases in dry weight, COD and EOX, compared to the control, were seen in sludges receiving bicarbonate; after 5 months the reductions were 60% (COD) and 50% (EOX). The results indicate that the use of bicarbonate to adjust pH could be advantageous in anaerobic treatment of organochloride-containing residues.

Laser-printed paper was treated individually and with combinations of purified endoglucanases from Gloeophyllum sepiarium (EGS) and Gloeophyllum trabeum (EGT), a xylanase from Thermomyces lanuginosus (X) and a mannanase from Sclerotium rolfsii (M). Subsequent toner removal efficiency after enzymatic and both magnetic or flotation deinking treatment was assessed by image analysis. The enzyme effect was more pronounced in magnetic deinking, demonstrating 94% removal of toner using a combination of EGS and X. The use of the pure EGT and EGS suggests that endoglucanases are responsible for most of the success in deinking. Compared to flotation deinking (89%), a higher yield of fibres (97.2%) was attained with this new combined enzymatic–magnetic technique, which consequently uses only tap water and no chemicals. While all enzyme applications increased freeness and slightly reduced intrinsic fibre strength, the resultant handsheets strength was maintained or marginally improved. The nature of the individual endoglucanase seems to play an important role since different deinking efficiencies and strength properties were measured with EGS and EGT, which are closely related enzymes in terms of molecular properties and specificities on model substrates.

The antioxidant and antimicrobial properties of Zataria multiflora and Iranian and European Cumin were evaluated, first as pure extracts and later as active agents incorporated in paper, manufactured at laboratory scale as an active packaging material. Two different procedures were used for quantifying the antioxidant properties and microbiological studies versus Staphylococcus aureus , Listeria innocua, Pseudomonas sp., Salmonella enterica subsp. enterica and Escherichia coli were carried out. All bacteria were inhibited when exposed to the atmosphere generated from 4% to 6% (w/w) of Zataria essential oil in the active coating. Zataria showed the best antioxidant properties. The compounds responsible for the antimicrobial and antioxidant properties are analyzed and discussed.

Huge numbers of cultural objects were damaged by microorganisms after having been soaked with seawater from the tsunami that followed the Great East Japan Earthquake on March 11, 2011. Among the cultural objects that were damaged, paper-based objects in the tsunami area were severely affected. In the present study, a culture-based analysis and on-site investigations were carried out to determine the extent of the microbial deterioration of paper-based objects (i.e., administrative documents and historical documents). Dark-colored fungi were isolated from black-spotted paper as Stachybotrys chartarum , which was observed to grow significantly in mineral salt medium supplemented with cellulose as the sole nutrient source; growth was shown in 10% NaCl. Two isolates from light red-spotted paper were related to Streptomyces sp. and Myxotrichum sp., respectively. These isolates generated red pigments in the mycelia and on the culture plates. Penicillium -related isolates were dominant in the sample tested, and they showed a higher level of NaCl tolerance. From the results of our on-site investigations, a unique discoloration, mainly black- and light red-spotted alterations caused by microbes, was observed on many paper-based objects that had been left wet for several months. The adhesive tape method applied to the black-spotted and red-spotted areas on damaged documents, Stachybotrys , Chaetomium and Cladosporium , and Actinomycetales bacterium were observed, respectively. With regard to the discoloration found on tsunami-affected paper-based objects, Stachybotrys and Streptomyces sp. seem to be responsible for the black- and red-spotted paper alterations, respectively.

A strain of Bacillus subtilis IARI-SP-1 isolated from soil long term irrigated with effluents of paper and pulp mill showed high β-1, 4-endoglucanase (2.5 IU/ml) but low activity of β-1, 4-exoglucanase (0.8 IU/ml) and β-glucosidase (0.084 IU/ml). The β-1, 4-endoglucanase gene of IARI-SP-1 was amplified using degenerate primers designed based on sequences already available in NCBI GenBank. A full length gene of β-1, 4-endonuclease consisting of 1499 nucleotides was identified through sequence analysis of the amplified product. The ORF encoded for a protein of 500 amino acids with a predicted molecular weight of 55 kDa. The gene was cloned in pET-28a and over expressed in Escherichia coli BL21 (DE3). In comparison to wild strain ( B. subtilis ), the transformed E. coli exhibited four times increase in cellulase production. Higher enzyme activity was observed in supernatant (8.2 IU/ml) than cell pellet (2.8 IU/ml) suggesting more extracellular production of β-1, 4-endoglucanase. SDS-PAGE and CMC plate assay also confirmed the overproduction by the transformed E. coli . The pH and temperature optima of expressed β-1, 4-endoglucanase enzyme was identical to that of wild strain and was 8 and 50–60 °C, respectively.

Fungal melanin staining is a problem on many cultural objects, ranging from the French Palaeolithic cave at Lascaux to books and papers in museum collections. Melanin, because it is insoluble and resistant to bleaching, may leave behind undesirable stains long after the fungal infestation has been controlled. Research into removal of melanin stains from paper and other sensitive substrates using industrial biomimetic oxidizing systems has shown considerable success. We studied relative concentration of the bleaching reagents and the reaction kinetics both in liquid suspensions of melanin and on melanized paper samples. Liquid suspension samples were tested for changes in their chemical composition (appearance and relative representation of functional groups and chemical bonds) with FTIR spectrometry. Changes in color of melanized paper samples were investigated with a CIE L*a*b system, where the effectiveness of the treatment (bleaching) was determined as a change in lightness (ΔL). Melanin was oxidized in the liquid suspensions, and the intensity of modification depended on the procedure employed. Bleaching of melanin with the biomimetic copper–pyridine complex proved to be far superior to the effect of white-rot fungal oxidizing enzymes, previously reported on by this group.

One of the most important causes of paper degradation is biodeterioration, caused especially by fungi and bacteria under certain environmental conditions. This study was designed to check the presence of cellulolytic microorganisms on documents from the 19th century, and to verify any biodeterioration that may have occurred by using a series of predominantly non-invasive analyses in order to preserve their integrity. For the characterization of paper documents various non-destructive micro-analyses were performed in succession, so as to deepen our knowledge of these step by step and thus provide a logical approach for the general study of this type of artifact. The samples were analyzed by using light microscopy, histochemical tests, pH analysis, SEM–EDS observations, Fourier transform infrared (FTIR) spectroscopy in ATR, and GC–MS. Organic cultures were also made with the samples taken from the surfaces of the documents; in particular, four fungal species, Aspergillus sp., Penicillium sp., Cladosporium sp., and Ulocladium alternarie, have been identified, as well as the presence of bacteria and dust mites. Finally, relations/correlations between the results of chemical and biological analyses are discussed.

The microbial diversity of different kinds of stains present on the surface of 14 historical documents (nine parchments and five paper letters) was evaluated through a combination of cultural and molecular methods. The samples were recovered using adhesive tape and swabs and were afterwards treated in two different ways: (1) direct inoculation on agar plates; or (2) suspension in physiological solution and plating in specific plates for the growth of bacteria and fungi. The isolated microorganisms, before identification, were selected by two different PCR-based methods – f-ITS and f-CBH, for bacteria and fungi, respectively. The f-ITS method is based on the amplification of the internal transcribed sequence between the bacterial 16S and 23S rDNA. The f-CBH method is a new molecular selection tool oriented to the fungal cellobiohydrolase gene. Both PCR selection methods produced typical profiles, which clustered the isolates in order to reduce them for subsequent sequencing identification through the amplification of the fungal 28S rRNA and the bacterial 16S rRNA genes. The cellulolytic and proteolytic abilities were screened through the use of three plate assays, the Ostazin Brilliant Red H-3B (OBR-HEC), milk agar, and gelatin agar. Massilia timonae , Lysobacter dokdonensis , and strains belonging to the genus Bacillus sp., Microbacterium sp., and Curtobacterium sp. with different fungal members such as Aspergillus fumigatus , Penicillium commune , Mucor spinosus , and Phoma herbarum (all recovered from paper) displayed both biodegradative activities. The parchment isolates with a marked proteolytic activity were Bacillus cereus , Staphylococcus epidermidis , Pseudomonas oryzihabitans , Virgibacillus sp., Micromonospora sp., and again members of the fungal genera Penicillium , Mucor , and Phoma .

The growth of molds on paper containing cellulose is a frequent occurrence when the level of relative air humidity is high or when books become wet due to water leaks in libraries. The aim of this study is to differentiate the bioreceptivity of different types of book paper for different fungi. Laboratory tests were performed with strains of Aspergillus niger, Cladosporium sp., Chaetomium globosum and Trichoderma harzianum isolated from books. Four paper types were evaluated: couche, Polen (offset), recycled and a reference paper containing only cellulose. The tests were carried out in chambers with relative air humidity of 95% and 100%. Mold growth was greatest in the tests at 100% relative humidity. Results of stereoscopic microscopy observation showed that Cladosporium sp. grew in 74% of these samples, A. niger in 75%, T. harzianum in 72% and C. globosum in 60%. In the chambers with 95% air humidity Cladosporium sp. grew in only 9% of the samples, A. niger in 1%, T. harzianum in 3% and C. globosum did not grow in any sample. The most bioreceptive paper was couche and the least receptive was recycled paper. The composition of the recycled paper, however, varies depending on the types of waste materials used to make it.

Melanin is chemically and by physical characteristics very similar to lignin, a major constituent of wood, and therefore ligninolytic enzymes of white-rot fungi were tested for their ability to selectively degrade melanin. Melanin degradation was studied both in liquid suspensions of melanin and on melaninised paper samples. Liquid suspension samples were tested for changes in their chemical composition (appearance and relative representation of functional groups and chemical bonds) with FTIR spectrometry. Changes in colour of melaninised paper samples were investigated with a colorimeter. Effectiveness of the treatment (bleaching) was determined as a change in lightness (ΔL). Melanin was oxidised in the liquid suspensions, and the intensity of modification varied depending on the procedure employed. The most pronounced changes in melanin were observed in laccase-1-hydroxybenzotriazole (HBT) treatment at heightened air pressure. The most prominent discoloration of the melaninised paper samples (and no visually detectable damage to the integrity of the paper) was, like in the case of the liquid suspensions, observed after laccase-HBT treatment.

With many important artistic works and documents made of paper, and thus susceptible to biodeterioration by fungi, research is required in an effort to replace toxic chemical products with other more benign ones. In this work the antifungal effect of methyl and propyl paraben mixtures at different concentrations was evaluated. The fungi used in the experiments were a Cladosporium species and Penicillium corylophilum, both of which are well known as paper-biodeteriorating fungi. The results demonstrate that a mixture of 0.5% methyl paraben and 1% propyl paraben, in 85% ethanolic solution, is the lowest concentration necessary to provide an efficient antifungal action. A deacidification agent, 5% calcium propionate, was added to this mixture to produce a multi-purpose formulation to treat acidification and fungal contamination of paper documents. Tests carried out on paper samples before and after application of this mixture showed only a minor increase in yellowing and a slight decrease in tensile strength, while substantially raising the pH, and thus the alkaline reserve, and also a slight increase in the percentage of deformation.

Biodeterioration is an undesirable process that can affect cultural heritage and economically important materials. Although several biotic and abiotic conditions can accelerate this process, microorganisms are perhaps its main promoters. Fungi are the most important microbial agents of biodeterioration of industrial paper stored in archives. The high genetic plasticity of these organisms allows them to adapt to different environments, using almost any class of materials as substrate. Fungi produce a wide array of enzymes, including cellulases, amylases, and proteases, which are responsible for their gross biodete- rioration activity. Thirty-two morphotypes of filamentous fungi were isolated on different media from industrial paper at an advanced stage of biodeterioration. The isolates showed different degrees of cellulolytic, proteolytic, and amylolytic activities on plate assays. The highest proteolytic and amylolytic activities were selected for isoform characterization, which provided an indication of the biochemical diversity that allowed them to colonize these materials. Eladia sacculum was the morphotype selected for partial purification of basic proteases since it has three basic isoforms, simplifying the purification process. We obtained a protein of 35 kDa with a pI of 8.9. 2008 Published by Elsevier Ltd.

Pentachlorophenol (PCP) degrading Bacillus strains CL3, CL5, and CL11 were isolated from the secondary sludge of a pulp paper mill and characterized. These isolates were identified as Bacillus megaterium , Bacillus pumilus , and Bacillus thuringensis based on their 16S rRNA sequence analysis. These isolates were able to grow and utilize PCP as a carbon and energy source. HPLC analysis and stoichiometric release of chloride in the medium confirmed the degradation ability of these isolates. The removal efficiency of PCP by these bacterial isolates was highly significant and they were able to degrade more than 90% of PCP when grown at high concentration of PCP (600 mg l −1 ). Consortia of these isolates removed 77% of PCP from the sludge. The removal efficiency of PCP by the bacterial isolate used in this study was found to be more efficient than what has been reported with other Bacillus species. These results suggest that the bacterial isolates are very effective PCP degraders and can be used in remediation of PCP-contaminated sites.

In this study, the changes in the physical (density and thickness swelling), mechanical (modulus of rupture, modulus of elasticity, internal bond), and biological (decay resistance) properties of particleboard dependence on the melamine paper weight, varnish type and the structure of continuous press laminate (CPL) were investigated. It was shown that coating of board surfaces and edges increased the mechanical properties except for internal bond strength, decreased the thickness swelling, and improved the decay resistance of particleboard, significantly. Melamine paper weight, the structure of CPL and varnish types were found to be effective on the some properties of particleboard.

The composition of filamentous bacteria in paper machine systems has received little scientific scrutiny, even though these organisms have been associated with many problems that affect machine efficiency and paper quality. The objective of the study was to isolate and characterize filamentous bacteria in paper mill slimes using conventional microbiological techniques, fatty acid methyl ester (FAME) analysis, and 16S rRNA gene sequencing. Filamentous or long, thread-like bacteria from different genera were observed. Pink or red-pigmented filamentous bacteria identified as Flectobacillus sp. were commonly isolated from pink slime deposits, but they were also obtained from deposits that did not appear pink or red. Two organisms had certain characteristics that were different from similar organisms previously described. One was a Gram-negative, filamentous bacterium with golden yellow colonies. This organism was esculin positive, and hydrolyzed starch but did not produce hydrogen sulfide. A BLAST search of GenBank database produced an identity of 92% with Cytophaga sp. or Flavobacterium columnare. A Gram-positive bacterium that produced very long or filamentous structures was also observed. On tryptic soy agar, this organism produced yellow colonies, but on plate count agar, the colonies were white. The plate count agar-grown cells were atypical with many of them having bulbs either in the middle or at the ends of the cell. In indole nitrite broth, the organism produced a very extensive filamentous structure. The FAME and 16S rRNA gene analyses of this organism showed that the organism was a Bacillus sp., but no spores were produced in any of the media studied, including a sporulating medium. Various spore-formers, identified as Bacillus psychrophilus and Paenibacillus sp., also had long continuous chains of cells. Two species of Chryseobacterium produced long filaments. Actinomycetes with branched mycelia and identified as Norcardiopsis alba and Streptomyces albidoflavus were isolated from wet lap pulp samples. These actinomycetes produced a very strong earthy odor both in culture and in field samples. Information, such as growth requirements and presence or absence of endospores, gained about these organisms will help in their control.

Pathogenic agents and pests attack human’s health and their wealth. They are living organisms that comply with taxonomy and ecology as all other living beings in the biosphere do. Their ability to succeed as pathogenic agents or pests seems to depend to a large extent upon their lifehistory traits, ecology, genetic background and ultimately upon their evolution. Altogether research seeking to understand the detailed interaction between parasites, bacteria, viruses and their potential human targets has to characterise all the organisms involved and, consequently, deal with the methods and approaches used in the fields of fundamental biological sciences, such as genetics, ecology, phylogenetics and taxonomy. In this context, it should be noted that quite often, research in pest management or pathology does not involve a basic, preliminary investigation of the fundamentals of the organisms involved in the phenomenon, but instead turns to interaction analysis. For instance, when it comes to virus– vector–host interaction and disease, it is amazing to note that the importance is given to not only an accurate determination of the virus responsible for an emerging disease but also the lack of interest in obtaining similar information leading to a precise identification of the reservoir and/or vector s.l. of the virus. Even in some, recent publications, a virus reservoir can be simplistically identified as a ‘‘rodent’’ or a ‘‘rat’’. Although hosts should be identified under their binomial scientific name, all too often they are acknowledged by an obsolete or synonymous name, or it is impossible to find the species as reported in the study area.

Books and documents are subject to decay by a variety of organisms, including mould. Experiments were conducted to find the lowest gamma radiation dose needed to inactivate fungi. Aspergillus versicolor and Eurotium chevalieri, previously isolated from naturally contaminated book from a Brazilian public library and from the environment, respectively, were cultivated and treated in a Co60 irradiation unit with doses ranging from 14.5 to 25 kGy. The minimum dose required to kill these fungi was 16 kGy. Thus, this dose was applied directly to a severely attacked book and deteriorated old documents. Pieces of damaged paper from these materials were incubated in culture media before and after the treatment. Several fungi were isolated and identified, including representatives of Acremonium, Aspergillus, Cladosporium, Fusarium, Penicillium and Trichosporon. After the treatment, no living fungi were detected from the irradiated material. The book was maintained in favorable conditions for new fungal attack for 2 months and no fungal growth was detected. These results are very promising and demonstrate the effectiveness of gamma-ray radiation for the recovery of severely damaged books and old documents, leading to the preservation of our cultural heritage and prevention of diseases caused by moulds in libraries and archives.