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The archive of the Universidad de Costa Rica maintains a nineteenth-century French collection of drawings and lithographs in which the biodeterioration by fungi is rampant. Because of nutritional conditions in which these fungi grew, we suspected that they possessed an ability to degrade cellulose. In this work our goal was to isolate and identify the fungal species responsible for the biodegradation of a nineteenth-century art collection and determine their cellulolytic activity. Fungi were isolated using potato-dextrose-agar (PDA) and water-agar with carboxymethyl cellulose (CMC). The identification of the fungi was assessed through DNA sequencing (nrDNA ITS and α-actin regions) complemented with morphological analyses. Assays for cellulolytic activity were conducted with Gram’s iodine as dye. Nineteen isolates were obtained, of which seventeen were identified through DNA sequencing to species level, belonging mainly to genera Arthrinium, Aspergillus, Chaetomium, Cladosporium, Colletotrichum, Penicillium and Trichoderma. For two samples that could not be identified through their ITS and α-actin sequences, a morphological analysis was conducted; they were identified as new species, named Periconia epilithographicola sp. nov. and Coniochaeta cipronana sp. nov. Qualitative tests showed that the fungal collection presents important cellulolytic activity.

Paper mulberry (Broussonetia papyrifera) is a well-known woody tree historically used for Cai Lun papermaking, one of the four great inventions of ancient China. More recently, Paper mulberry has also been used as forage to address the shortage of feedstuff because of its digestible crude fiber and high protein contents. In this study, we obtained a chromosome-scale genome assembly for Paper mulberry using integrated approaches, including Illumina and PacBio sequencing platform as well as Hi-C, optical, and genetic maps. The assembled Paper mulberry genome consists of 386.83 Mb, which is close to the estimated size, and 99.25% (383.93 Mb) of the assembly was assigned to 13 pseudochromosomes. Comparative genomic analysis revealed the expansion and contraction in the flavonoid and lignin biosynthetic gene families, respectively, accounting for the enhanced flavonoid and decreased lignin biosynthesis in Paper mulberry. Moreover, the increased ratio of syringyl-lignin to guaiacyl-lignin in Paper mulberry underscores its suitability for use in medicine, forage, papermaking, and barkcloth making. We also identified the root-associated microbiota of Paper mulberry and found that Pseudomonas and Rhizobia were enriched in its roots and may provide the source of nitrogen for its stems and leaves via symbiotic nitrogen fixation. Collectively, these results suggest that Paper mulberry might have undergone adaptive evolution and recruited nitrogen-fixing microbes to promote growth by enhancing flavonoid production and altering lignin monomer composition. Our study provides significant insights into genetic basis of the usefulness of Paper mulberry in papermaking and barkcloth making, and as forage. These insights will facilitate further domestication and selection as well as industrial utilization of Paper mulberry worldwide.

This article presents a novel method for integrating native corn starch and calcium carbonate (CaCO3) into a hybrid pigment, which can be applied in papermaking to increase the amount of starch in paper and improve sheet bonding without impairing dewatering. Starch was first thermally treated to partially gelatinize and swell the granules. The aim of the treatment was to change the surface properties of starch and thus to improve the interaction with CaCO3 particles. CaCO3 shell was precipitated around the swollen granules and the encapsulated starch was then cooked to achieve complete gelatinization. When the granules were cooked, the starch became active for bonding, but its solubility was very limited due to the encapsulation. Particle size measurement and scanning electron microscopy were used to analyze the sample properties during the preparation. The hybrid pigment was added to paper sheets and its effect on the paper properties was examined. Starch was shown to be effectively bound to the pigment structure. The hybrid pigment allowed a large amount of native corn starch to be added to paper without interfering with the papermaking process. Furthermore, it gave excellent mechanical paper properties.

International audience; We report a pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) method capable of analyzing traditional East Asian papers. The method proposed is based on rapid and easy single step Py-GC/MS analysis that can be carried out with a minimum amount of matter, in the few g range. Three reference papers manufactured from kozo (Broussonetia kazinoki Siebold & Zucc.), mitsumata (Edgeworthia chrysantha Lindl.) and gampi (Wikstroemia sikokiana Franch. & Sav.) with the traditional hand paper making processes were examined. The method allows discrimination between terpenic and steroid compounds, which were revealed as chemical markers of origin of the plant fibers. Each paper investigated was found to have characteristic pyrolysis fingerprints that were unique to the traditional handmade paper, demonstrating the potential for differentiation of these biochemical components of fiber plants on East Asian papers towards identification and conservation of cultural heritage. The investigation on Py-GC/MS was extended to liquid extraction followed by GC/MS analysis to characterize the biochemical components of fiber plants. The main contribution of this study is to provide molecular criteria for discriminating plant species used for traditional East Asian hand papermaking. Py-GC/MS complements efficiently microscope identification especially for adverse cases. A case study of archaeological Chinese paper painting artefacts was thereafter successfully investigated to address informative potential and efficiency of the criteria of identification on ancient and degraded East Asian paperworks.

The competence of novel fungal consortium, consisting of Nigrospora sp. LDF00204 (accession no. KP732542) and Curvularia lunata LDF21 (accession no. KU664593), was investigated for the treatment of pulp and paper mill effluent. Fungal consortium exhibited enhanced biomass production under optimized medium conditions, i.e., glucose as carbon (C), sodium nitrate as nitrogen (N), C/N 1.5:0.5, pH 5, temperature 30 °C, and agitation 140 rpm, and significantly reduced biochemical oxygen demand (85.6%), chemical oxygen demand (80%), color (82.3%), and lignin concentration (76.1%) under catalytic enzyme activity; however, unutilized ligninolytic enzymes, such as laccase (Lac), manganese peroxidase (MnP), and lignin peroxidase (LiP), were observed to be 13.5, 11.4, and 9.4 U/ml after the third cycle of effluent treatment in repeated batch process. Scanning electron microscopy (SEM) of fungal consortium revealed their compatibility through intermingled hyphae and spores, while the FTIR spectra confirmed the alteration of functional groups ensuring structural changes during the effluent treatment. Gas chromatography/mass spectroscopy (GC-MS) analysis showed the reduction of complex compounds and development of numerous low-molecular-weight metabolites, such as 1-3-dimethyl benzene, 2-chloro-3-methyl butane, pentadecanoic acid, and 1-2-benzene dicarboxylic acid, during the treatment, demonstrating the massive potential of the novel fungal consortium to degrade recalcitrant industrial pollutants.

Peaches are known for their palatable flavor and abundant nutrients. However, peaches are perishable, and the existing preservation techniques for peaches are still immature. To further extend the shelf life and prevent nutrient loss of perishable peaches under ambient temperature in summer (approximately 25–32 °C), we conducted experiments wrapping peaches (Prunus persica cv ‘Baihua’) in single- and composite-treated vegetal fibrous papers that contained calcium carbonate, phytic acid, Na-alginate and vitamin C. The pathogenic fungi that primarily caused peach decay during storage belonged to the genera of Penicillium, Botrytis, Aspergillus, Alternaria, and Rhizopus. After analyzing quality attributes, including weight loss, firmness, soluble sugar content, respiration rate, relative electric conductivity, malonaldehyde content, peroxidase activity and the decay index, we proved that vitamin C within the preservative paper greatly contributes to peach preservation. Combined with phytic acid and Na-alginate, the composite vitamin C preservative papers played significant roles in delaying fruit senescence, and 0.4% (w/v) vitamin C preservative paper with 1% Na-alginate could maintain quality and extend shelf life with the best effect. This preservation technique significantly postponed the respiration peak by 2–3 days and is a significant contribution to contemporary commercial production.

Past studies have revealed major difficulties in applications of xylanase in the pulp and paper industry as enzymes isolated from many different species could not tolerate high temperatures or highly alkaline conditions. The thermostable xylanase A from Bacillus halodurans C-125 (C-125 xylanase A) was successfully cloned and expressed in Pichia pastoris with a yield as high as 3361 U/mL in a 2 L reactor. Its thermophilic and basophilic properties (optimal activity at 70 °C and pH 9.0), together with the fact it is cellulase-free, render this enzyme attractive for compatible applications in the pulp and paper industry. The pretreatment of wheat straw pulp with C-125 xylanase A at pH 9.0 and 70 °C for 90 min induced the release of both chromophores (Ab(237), Ab(254), Ab(280)) and hydrophobic compounds (Ab(465)) into the filtrate as well as sugar degradation. Moreover, the addition of 10 U xylanase to 1 g wheat straw pulp (dry weight) as pretreatment improved brightness by 5.2% ISO and decreased the kappa number by 5.0% when followed by hydrogen peroxide bleaching. In addition, compared with two commercial enzymes, Pulpzyme HC and AU-PE89, which are normally incorporated in ECF bleaching of wheat straw pulp, C-125 xylanase A proved to be more effective in enhancing brightness as well as preserving paper strength properties. When evaluating the physical properties of pulp samples, such as tensile index, tearing index, bursting index, and post-color (PC) number, the enzymes involved in pretreating pulps exhibited better or the same performances as chemical treatment. Compared with chemical bleaching, chlorine consumption can be significantly reduced by 10% for xylanase-pretreated wheat straw pulp while maintaining the brightness together with the kappa number at the same level. Scanning electron microscopy revealed significant surface modification of enzyme-pretreated pulp fibers with no marked fiber disruptions.

Pulp mill wastewater generated from wheat straw is characterized as high alkalinity and very high COD pollution load. A naturally developed microbial community in a pulp mill wastewater storage pool that had been disused were investigated in this study. Owing to natural evaporation and a huge amount of lignocellulose's deposition, the wastewater sediment contains high concentrations of organic matters and sodium ions, but low concentrations of chloride and carbonate. The microbiota inhabiting especially anaerobic community, including methanogenic arhcaea and cellulolytic species, was studied. All archaeal sequences fall into 2 clusters of family Halobacteriaceae and methanogenic archaeon in the phylum Euryarchaeota. In the methanogenic community, phylogenetic analysis of methyl coenzyme M reductase A (mcrA) genes targeted to novel species in genus Methanoculleus or novel genus of order Methanomicrobiales. The predominance of Methanomicrobiales suggests that methanogenesis in this system might be driven by the hydrogenotrophic pathway. As the important primary fermenter for methane production, the cellulolytic community of enzyme GHF48 was found to be dominated by narrower breadth of novel clostridial cellulase genes. Novel anoxic functional members in such extreme sediment provide the possibility of enhancing the efficiency of anoxic treatment of saline and alkaline wastewaters, as well as benefiting to the biomass transformation and biofuel production processes.

Mangrove fungi are vastly unexplored for enzymes with industrial application. This study aimed to assess the biocatalytic activity of mangrove fungal xylanases on recycled paper pulp. Forty-four mangrove fungal (MF) isolates were initially screened for xylanolytic activity in minimal medium with corn cob xylan as the sole carbon source. Eight MF were further cultivated under submerged fermentation for the production of crude xylanases. These crude enzymes were then characterized and tested for the pretreatment of recycled paper pulps. Results showed that 93 % of the tested MF isolates exhibited xylanolytic activity in solid medium. In submerged fermentation, salinity improved the growth of the fungal isolates but did not influence xylanase production. The crude xylanases were mostly optimally active at 50 °C and pH 7. Changes in pH had a greater effect on xylanase stability than temperature. More than half of the activity was lost at pH 9 for majority of the crude enzymes. However, two thermophilic xylanases from Fusarium sp. KAWIT-A and Aureobasidium sp. 2LIPA-M and one alkaliphilic xylanase from Phomopsis sp. MACA-J were also produced. All crude enzymes exhibited cellulase activities ranging from 4 to 21 U/ml. Enzymatic pretreatment of recycled paper pulps with 5 % consistency produced 70-650 mg of reducing sugars per gram of pulp at 50 °C after 60 min. The release of high amounts of reducing sugars showed the potential of mangrove fungal crude xylanases in the local paper and pulp industry. The diverse properties shown by the tested crude enzymes also indicate its potential applications to other enzyme-requiring industries.

Leucaena leucocephala is a leguminous tree species accounting for one-fourth of raw material supplied to paper and pulp industry in India. Cinnamate 4-Hydroxylase (C4H, EC 1.14.13.11) is the second gene of phenylpropanoid pathway and a member of cytochrome P450 family. There is currently intense interest to alter or modify lignin content of L. leucocephala. Three highly similar C4H alleles of LlC4H1 gene were isolated and characterized. The alleles shared more than 98 % sequence identity at amino acid level to each other. Binding of partial promoter of another C4H gene LlC4H2, to varying amounts of crude nuclear proteins isolated from leaf and stem tissues of L. leucocephala formed two loose and one strong complex, respectively, suggesting that the abundance of proteins that bind with the partial C4H promoter is higher in stem tissue than in leaf tissue. Quantitative Real Time PCR study suggested that among tissues of same age, root tissues had highest level of C4H transcripts. Maximum transcript level was observed in 30 day old root tissue. Among the tissues investigated, C4H activity was highest in 60 day old root tissues. Tissue specific quantitative comparison of lignin from developing seedling stage to 1 year old tree stage indicated that Klason lignin increased in tissues with age.

A lignin peroxidases-producing Serratia liquefaciens was used for bioremediation of pulp and paper (P&P) mill effluent. The treatment led to reduction of chemical oxygen demand (COD), colour, lignin and phenolic content by 84%, 72%, 61% and 95%, respectively. The effluent detoxification was studied by genotoxicity assays using Allium cepa L. (onion) root tip cells. Genotoxicity studies included measuring mitotic index (MI), chromosomal aberrations (CA) and nuclear abnormalities (NA) in root tip cells following treatment with 25, 50, 75 and 100% (v/v) of effluent. The root tip cells grown in untreated effluent showed a significant decrease in MI from 69% (control) to 32%, 27%, 22% and 11% at 25%, 50%, 75% and 100% effluent concentration, respectively. This indicated that the untreated effluent was highly cytotoxic in nature. Further, root tip cells, when treated with different concentrations of effluent showed various CA and NA including c-mitosis, stickiness, chromosome loss, chromosome break, anaphase bridge, multipolar anaphase, vagrant chromosomes, micronucleated and binucleated cells. The MI observed in root tip cells grown in bacterial treated effluents at similar concentrations (25, 50, 75 and 100% v/v) showed an increase of 33%, 36%, 42% and 66%. CA showed a substantial decrease and in some instances, complete absence of CA was also observed. The findings suggest that S. liquefaciens culture could be a potential bacterial culture for bioremediation of P&P mill effluent, as it is effective in substantial lowering of pollutants load as well as reduces the cytotoxic and genotoxic effects of effluent.

The study was conducted with an aim to remediate effluent from a pulp and paper mill, after treating it for 28 days with an aquatic fern, Salvinia cucullata. The effluent had high BOD, COD, TS, TSS, TDS, P, hardness and chloride, and several heavy metals (Cd, Cu, Cr, Ni, Pb, Mg, Mn, Fe and Zn) above national limits. However, the plant survived a wide range of effluent concentrations (25%, 50%, 75% and 100%, v/v), and flourished well, particularly at 25% (v/v), resisted membrane injury and generation of H2O2 and O2, showed better growth and induced all the major antioxidant enzymes. The plants also induced lipid peroxidation. Most of the elemental profiles were higher than the toxic levels stipulated for plants, indicating tolerance to metal. In fact, barring Fe, for Cr, Cu, Pb, Zn, Mg and P, at all the effluent doses, and for Cd, Ni and Mn, up to 75% (v/v) effluent, greater concentrations were observed in leaf than in root. This plant was more suited for nutrient removal, as it effectively reduced BOD, Zn, Fe, Ni, Mg, P and increased dissolve oxygen. Further, pH, hardness, chloride, TS and Mn was reduced optimally by 25–50% (v/v) treatments. SEM revealed prominent structural damages from 50 to 100% treatments. Presence of Pb as well as Fe in the EDX peaks were observed in the cortex rather than in the root vascular zone. This plant could be suggested to be an effective phytoremediator of multi-contaminant effluent with maximum benefit at low doses (25–50%, v/v).

The study was conducted to evaluate the quality of dissolving pulp of Musa sapientum L. (banana) plant stem and its potential for biorefinery. Introduction of pre-hydrolysis prior to any alkaline pulping process helps to reduce the content of hemicellulose and consequently produce acceptably high content of cellulose pulp. Water pre-hydrolysis was done at 150 °C for 90 min. The amount of lignin, xylan and glucan in the extracted pre-hydrolysis liquor (PHL) was 1.6, 4.9 and 1.6%, respectively. Pulping of pre-extracted chips was done following soda-AQ, alkaline sulfite and kraft process. The ratio of chip to liquor was 1:7 for both pre-hydrolysis and pulping. The kraft pulping process with 20% active alkali and 25% sulfidity at 150 °C for 90 min showed the best result. The lowest kappa number was 26.2 with a considerable pulp yield of 32.7%. The pulp was bleached by acidic NaClO2 and the consistency was 10% based on air-dried pulp. The lowest amount of 7% NaClO2 was used for the bleaching sequence of D0ED1ED2. After D0ED1ED2 bleaching, the pulp showed that α-cellulose, brightness and ash were 91.9, 77.9 and 1.6% respectively. The viscosity was 19.9 cP. Hence, there is a possibility to use banana plant stem as a raw material for dissolving grade pulp and other bioproducts.

Artículo científico Trametes versicolor and Pleurotus ostreatus were used for the biopulping from pineapple leaf fiber (PALF). PALF substrate was subjected to T. versicolor for 2 to 6 weeks and to P. ostreatus for 4 to 8 weeks. The yields, holocellulose and lignin contents, and extractives in ethanol-toluene mixture and in sodium hydroxide (NaOH) solution were evaluated. Fourier transform infrared spectroscopy (FTIR) spectra, thermogravimetric analysis (TGA), and color studies by L*a*b* systems were used for sample analysis. The results showed that the pulp yield was 55% to 70% with P. ostreatus and 35% to 50% with T. versicolor. Longer colonization periods increased the amount of holocellulose and decreased the amount of lignin and extractives in ethanol-toluene and NaOH solution. TGA showed an increase in intensity associated with cellulose, and the observed inflexion was attributed to lignin, which showed a tendency to fade. The FTIR spectrum showed high intensity between 3100 cm-1 and 3600 cm-1 (cellulose) and decreased intensity at 1730 cm-1 (lignin). For both fungi, the pulp color produced an increase in L* color parameter and decreased in yellowness, while little variation was observed in redness. The most appropriate colonization period was 5 weeks for P. ostreatus and 4 weeks for T. versicolor.

O objetivo deste estudo foi avaliar as propriedades de polpas kraft marrons de madeiras de Eucalyptus grandis e Eucalyptus urograndis de composição química distinta, principalmente em relação aos seus conteúdos de xilanas, para fabricação de papéis. As polpas marrons de número kappa 17-18 e conteúdos de xilanas variando de 6 a 21% foram produzidas sob diferentes protocolos de cozimento: 1) kraft convencional; 2) pré-hidrólise kraft; 3) kraft de alta alcalinidade; e 4) kraft de alto rendimento. As polpas marrons foram refinadas em moinho PFI e seus respectivos papéis produzidos foram avaliados quanto às suas propriedades físico-mecânicas e ópticas. De modo geral, as propriedades de resistência dos papéis foram beneficiadas pelo conteúdo de xilanas. Para níveis muito altos de refino, ocorreram quedas bruscas de resistência ao rasgo para os papéis e o consumo de energia de refino foi influenciado pelo conteúdo de xilanas nas polpas marrons. Os mais altos valores de volume específico aparente foram obtidos para os papéis com baixo conteúdo de xilanas, o que os torna interessantes, se branqueados, no segmento de papéis tissue. Os papéis com elevado conteúdo de xilanas apresentaram-se economicamente atrativos e com características de resistências adequadas para o segmento de papéis de imprimir e escrever. The objective of this study was to evaluate properties of brown pulp from Eucalyptus grandis and Eucalyptus urograndis of distinct chemical composition, mainly in refers to its xylans content, for the manufacture of tissue and printing and writing (P&W) paper grades. Brow pulps of kappa number 17-18 and xylans contents varying 6-21% had been produced under different cooking protocols: 1) conventional kraft; 2) pre-hydrolysis kraft; 3) high alkalinity kraft; and 4) kraft of high yield. The brown pulps had been beated in PFI mill and its respective produced papers had been evaluated as regards physical-mechanical and optic properties. The highest bulk values had been gotten for papers with low xylans contents, what had revealed interesting, if bleached, in the manufacture of tissue papers. The papers with high xylans contents, had been presented technically viable and with resistance characteristics adjusted for the manufacture of P&W papers.

Dehydration-responsive element binding/C-repeat-binding factors (DREB/CBF) control the activity of multiple stress response genes and therefore represent attractive targets for genetic improvement of abiotic stress tolerance. Paper mulberry (Broussonetia papyrifera L. Vent) is well known for its bark fibers and high levels of chalcone and flavonoid derivatives. Transgenic paper mulberry plants expressing a tall fescue (Festuca arundinacea Schreb.) FaDREB1 gene under the control of CaMV 35S were produced to examine the potential utility of FaDREB1 to increase the tolerance of paper mulberry plants to abiotic stress. The overexpressing FaDREB1 plants showed higher salt and drought tolerance than the wild-type plants (WT). After 13 days of withholding water, or 15 days in the presence of 250 mM NaCl, all the WT plants died, while the over-expressing FaDREB1 plants survived. The FaDREB1 plants had higher leaf water and leaf chlorophyll contents, accumulated more proline and soluble sugars, and had less ion leakage (which reflects membrane damage) than the WT plants had under high salt- and water-deficient conditions. The 35S promoter-driven expression of FaDREB1 did not cause growth retardation under normal growth conditions. Therefore, improved tolerance to multiple environmental stresses in paper mulberry might be achieved via genetic engineering through the ectopic expression of an FaDREB1 gene.

In this work, we applied scanning electron microscopy (SEM), microanalysis and Raman spectroscopy to study the fungi inhabiting a richly illuminated parchment document and the damage induced by their activity. To that aim, we collected samples of fungal mycelium from the deteriorated areas on a removable adhesive tape specifically intended for lifting fungi without damaging the support. SEM analysis of the adhesive tape samples showed the co-occurrence of several species of fungi. One strain closely resembling Acremonium species was observed only in the tape micrographs but no agar cultures were obtained. Its fungal structures showed the production of abundant oxalates with an outstanding leaching of the calcium-based materials of parchment (typically manufactured with gypsum and lime). Needle-like crystals of calcium oxalate produced by the fungus forming a uniform and quite regular grid around conidial slimy heads were documented. As a result, the areas affected by moulds were weakened, stained and characterised by a powdery patina rich in calcium. Confocal μ-Raman confirmed the presence of oxalates while EDS showed the presence of calcium in crystals. We conclude that the defacement of the parchment was due to both collagenolytic activity, and to the biotransformation of calcium-based minerals by fungi.

Pretreatment of wheat straw pulp using cellulase-free xylanase produced from Bacillus stearothermophilus SDX at 60°C for 120min resulted in 4.75% and 22.31% increase in brightness and whiteness, respectively. Enzyme dose of 10U/g of oven dried pulp at pH 9 decreased the kappa number and permanganate number by 7.14% and 5.31%, respectively. Further chlorine dioxide and alkaline bleaching sequences (CDED(1)D(2)) resulted in 1.76% and 3.63% increase in brightness and whiteness, respectively. Enzymatic prebleaching of pulp decreased 20% of chlorine consumption without any decrease in brightness. Improvement in various pulp properties like viscosity, burst factor, burstness, breaking length, double fold, gurley porosity, tear factor, and tearness were also observed after bleaching of xylanase treated wheat straw pulp.

Two novel cellulase-poor xylanases from Coprinellus disseminatus SH-1 NTCC-1163 (enzyme-A) and SH-2 NTCC-1164 (enzyme-B) produced under solid-state fermentation mitigated kappa number of wheat straw soda-AQ pulps by 24.38 and 27.94% respectively after XE stages. The release of reducing sugars and chromophores was highest for both the enzymes at 10IU/g and reaction time 180min for 55°C at variable consistencies that is, 10% for enzyme-A and 5% for enzyme-B. (A)XECEHH and (B)XECEHH sequences improved brightness by 5.17 and 2.58% respectively at 4.5% chlorine charge. AOX in (A)XECEHH and (B)XECEHH sequences reduced by 56.11 and 55.75% respectively at 4.5% chlorine charge and 68.34 and 67.98% respectively at 2.25% chlorine charge respectively compared to control. Both the enzymes showed improvement in double fold and tear index with a decrease in burst and tensile index. SEM showed peeling, cracking and delamination in fibers due to enzyme treatment thus facilitating the penetration of bleach chemicals.

The genetic and functional diversity of Bacillus and Bacillus-derived genera was analyzed in soil samples collected from three different fields near Century Paper Mill, Lal Kuan, Uttarakhand, India. Two had been subjected to concentrated and diluted effluent irrigation for the past 25 years and were designated as a concentrated effluent irrigated field (CEIF) and a dilute effluent irrigated field (DEIF), respectively. The field irrigated with fresh water was designated as a water irrigated field (WIF). Increase in pH, Na and Zn content and decrease in Fe content was observed due to effluent irrigation. The population count of Bacillus and Bacillus-derived was maximum in DEIF followed by WIF and CEIF. Variations in plant growth-promoting traits and extracellular enzymes were observed among the isolates from the three different field soils. Based on the amplified ribosomal DNA restriction analysis (ARDRA) with three restriction enzymes, all the selected 104 isolates were clustered into 14 groups. The sequencing of the representative isolates revealed that the majority belonged to the genus Bacillus, while three isolates belonged to Paenibacillus, Lysinibacillus and Orthinibacillus. There were a few species like Orthinibacillus contaminans, B. oleronius, B. safensis, B. methylotrophicus, B. stratosphericus, B. aryabhattai, B. asahii and B. bataviensis that were prevalent only in DEIF and CEIF but not in WIF field soil. The diversity index parameters like the Shannon Index, indices of species richness and species evenness based on biochemical profiling and ARDRA profiling revealed that Bacillaceae members in the fresh water irrigated field were diverse.

Tagasaste wood (Chamaecytisus proliferus L.F. ssp palmensis) was characterized, chemical and energy terms, and assessed its potential as a lignocellulosic raw material, and its integral fractionation by autohydrolysis and delignification with ethanol. The hydrothermal treatment of the raw material at 175–185 °C provided a liquor containing a substantially increased amount of oligomers (between 16.6% and 47.4% as percentages with respect to the content of the raw material in each polymer fraction). A cellulose pulp with autohydrolysis treatment increased yields (53–60%), reduced Kappa number (28.8–34.6) but also viscosity (755–857 mL/g), and decreased paper strength (2.97–5.22 kN m/kg). However, beating ethanol cellulose from tagasaste was found to improve its strength-related properties more markedly than in soda pulp from the same material (tensile index of 44 kN m/kg).

Paper mills processing recycled paper suffer from biofouling causing problems both in the mill and final product. The total bacterial community composition and identification of specific taxa in the process water and biofilms at the stock preparation and paper machine areas in a mill with recycled paper pulp was described by using a DNA-based approach. Process water in a similar mill was also analyzed to investigate if general trends can be found between mills and over time. Bacterial community profiles, analyzed by terminal-restriction fragment length polymorphism (T-RFLP), in process water showed that the dominant peaks in the profiles were similar between the two mills, although the overall composition was unique for each mill. When comparing process water and biofilm at different locations within one of the mills, we observed a separation according to location and sample type, with the biofilm from the paper machine being most different. 16S rRNA gene clone libraries were generated and 404 clones were screened by RFLP analysis. Grouping of RFLP patterns confirmed that the biofilm from the paper machine was most different. A total of 99 clones representing all RFLP patterns were analyzed, resulting in sequences recovered from nine bacterial phyla, including two candidate phyla. Bacteroidetes represented 45% and Actinobacteria 23% of all the clones. Sequences with similarity to organisms implicated in biofouling, like Chryseobacterium spp. and Brevundimonas spp., were recovered from all samples even though the mill had no process problems during sampling, suggesting that they are part of the natural paper mill community. Moreover, many sequences showed little homology to as yet uncultivated bacteria implying that paper mills are interesting for isolation of new organisms, as well as for bioprospecting.

Paper documents in archives, libraries, and museums often undergo biodeterioration by microorganisms. Fungi and less often bacteria have been described to advance paper staining, so called “foxing” and degradation of paper substrates. In this study, for the first time, the fungal and bacterial diversity in biodeteriorated paper documents of Hellenic General State Archives dating back to the 19th and 20th century has been assessed by culture‐dependent and independent methods. The internally transcribed spacer (ITS) region and 16S rRNA gene were amplified by PCR from fungal and bacterial isolates and amplicons were sequenced. Sequence analysis and phylogeny revealed fungal phylotypes like Penicillium sp., Cladosporium sp., Penicillium citrinum, Alternaria infectoria, Alternaria alternata, Epicoccum nigrum, and Penicillium chrysogenum which are often implicated in paper deterioration. Bacterial phylotypes closely related to known biodeteriogenic bacteria such as Bacillus spp., Micrococcus spp., Kocuria sp. in accordance with previous studies were characterized. Among the fungal phylotypes described in this study are included well‐known allergens such as Penicillium spp., Alternaria spp., and Cladosporium spp. that impose a serious health threat on staff members and scholars. Furthermore, fungal isolates such as Chalastospora gossypii and Trametes ochracea have been identified and implicated in biodeterioration of historical paper manuscripts in this study for the first time. Certain new or less known fungi and bacteria implicated in paper degradation were retrieved, indicating that particular ambient conditions, substrate chemistry, or even location might influence the composition of colonizing microbiota.

It is generally accepted that dead tree decomposition is performed mainly by delignifying basidiomycetes. While ascomycetes have been reported to inhabit dead tree bark, their contribution to dead tree decomposition is still unclear. Here, we isolated five bark-inhabiting ascomycetes possessing cellulolytic activity from dead beech tree and assessed their polysaccharolytic activities. When cultivated in a medium containing filter paper as a sole carbon source, three strains degraded >40 % of the filter paper in a 4-week cultivation and the others degraded 15-30 % of the paper. The degraders possessed amylolytic, pectinolytic, and mannanolytic activities as well as cellulolytic activity, implying that they play an important role in dead tree decomposition after delignification by basidiomycetes. Phylogenetic analysis based on large subunit ribosomal DNA (lsu-DNA) sequences implied that the isolates belonged to Penicillium or Amorphotheca.

In this study geno-toxicity analysis along with effluent treatment was taken up to evaluate the efficiency of biological treatment process for safe disposal of treated effluent. Four fungi were isolated from sediments of pulp and paper mill in which PF4 reduced colour (30%) and lignin content (24%) of the effluent on 3rd day. The fungal strain was identified as Emericella nidulans var. nidulans (anamorph: Aspergillus nidulans) on the basis of rDNA ITS1 and rDNA ITS2 region sequences. The process of decolourization was optimized by Taguchi approach. The optimum conditions were temperature (30-35 degrees C), rpm (125), dextrose (0.25%), tryptone (0.1%), inoculum size (7.5%), pH (5) and duration (24h). Decolourization of effluent improved by 31% with reduction in colour (66.66%) and lignin (37%) after treatment by fungi in shake flask. Variation in pH from 6 to 5 had most significant effect on decolourization (71%) while variation in temperature from 30 to 35 degrees C had no effect on the process. Treated effluent was further evaluated for geno-toxicity by alkaline single cell gel electrophoresis (SCGE) assay using Saccharomyces cerevisiae MTCC 36 as model organism, indicated 60% reduction.

The influence of the hydrothermal treatment conditions of rice straw (temperature--150-190 degrees C , time--0-20 min after reaching the operation temperature, and liquid/solid ratio--6-8), on the composition of liquid and solid fractions was studied. Polynomial models were found for to reproduce the experimental results with errors less than 6%. Operating to 190 degrees C, 15 min and 9 liquid/solid ratio was obtained high values of glucose (1.92 g/L), xylose (3.97 g/L), arabinose (0.99 g/L) and acetic acid (1.96 g/L), what allows to save capital when not operating with the maximum time and/or liquid/solid ratio; yield in the solid fraction is 88.1% and lignin content 24.43%. If what it is required to obtain good values of yield (96.46%) and lignin content, is necessary to operate to 150 degrees C, obtaining lower concentrations glucose (1.11%), xylose (2.78%), arabinose (0.56%) and acetic acid (0.39%). Ethanolamine pulping of solid fraction of hydrothermal treatment provides pulp with higher or similar properties to those obtained of pulping rice straw with ethanolamine or soda.

Three white-rot fungi (Pleurotus sajor caju, Trametes versicolor and Phanerochaete chrysosporium) and one soft-rot fungi (Rhizopus oryzae) species confirmed their potential for future applications in the biological treatment of effluents derived from the secondary treatment of a bleached kraft pulp mill processing Eucalyptus globulus. Among the four species P. sajor caju and R. oryzae were the most effective in the biodegradation of organic compounds present in the effluent, being responsible for the reduction of relative absorbance (25-46% at 250 nm and 72-74% at 465 nm) and of chemical oxygen demand levels (74 to 81%) after 10 days of incubation. Laccase (Lac), lignin (Lip) and manganese peroxidases (MnP) expression varied among fungal species, where Lac and LiP activities were correlated with the degradation of organic compounds in the effluent treated with P. sajor caju. The first two axes of a principal component analysis explained 88.9% of the total variation among sub-samples treated with the four fungus species, after different incubation periods. All the variables measured contributed positively to the first component except for the MnP enzyme activity which was the only variable contributing negatively to the first component. Absorbances at 465 nm, LiP and Lac enzyme activities were the variables with more weight on the second component. P. sajor caju revealed to be the only species able to perform the biological treatment without promoting an increment in the toxicity of the effluent to the Vibrio fischeri, as it was assessed by the Microtox assay. The opposite was recorded for the treatments with the other three species of fungus. EC(50-5 min) values ranging between 28 and 57% (effluent concentrations) were recorded even after 10 to 13 days of treatment with P. chrysosporium, R. oryzae or with T. versicolor.

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.

White-rot fungus Trametes hirsute ( T.h. 19-6) was used for modifying the eucalyptus CTMP fibres. Results show that the T.h. 19-6 removed mainly lignin (by 7.42%) and extractive (by 11.52%) after a short period of 5 days incubation. Due to the fungal degradation of the fibre wall materials, the middle lamella remainder on the fibre surface was significantly reduced and the fibre wall structure was loosened, which led to an increase in fibre internal bonding strength by 32% and an increase in handsheet tensile index and tear index by 49%, and 34%, respectively. A subsequent PFI refining process further amplified the fungal treatment effect, resulting in extensive fibre internal fibrillation. Compared with untreated pulp, fungus-treated pulp reached the same freeness levels with less PFI revolutions, indicating potential energy saving in the refining process.

In this work, depithed cardoon stalk (Cynara cardunculus) has been used with the objective of obtaining bleachable pulps. The material, once properly prepared, was subjected to one-step peroxyformic acid delignification. In order to study the process, a face-centred second order factorial design was developed which allowed the determination of the influences of four variables: concentrations of formic acid and hydrogen peroxide in the cooking liquor and the time and temperature of the treatment. Empirical mathematical models have been obtained which predict the yield, kappa index, residual lignin content, and viscosity of the pulps. These models demonstrate that in general the delignification was extensive, producing pulps with kappa indexes less than 25 in the majority of cases, with good yields in the range of 45-60%. However, the pulps seem to have been degraded in the reaction media, as can be deduced from the low viscosity values found: 260-520 mL/g.

A Gram-positive, non-motile, rod-shaped, non-spore-forming bacterium, designated CAAS 251T, was isolated from paper-mill effluent in Wuhan, China. The organism grew optimally at 40-42 degrees C and at pH 9.0-10.0. The major menaquinones were MK-7, MK-8 and MK-9. The predominant cellular fatty acids were anteiso-C15:0 (34.78 %), anteiso-C17:0 (25.24 %) and C16:0 (13.37 %). The G+C content of the genomic DNA was 65.5 mol%. A phylogenetic analysis based on 16S rRNA gene sequences showed that strain CAAS 251T belongs to the genus Nesterenkonia, having sequence identities ranging from 96.0 to 97.0 % with respect to eight recognized species of the genus Nesterenkonia. Data from DNA-DNA hybridization and physiological and biochemical tests indicated that strain CAAS 251T represents a novel species of the genus Nesterenkonia, for which the name Nesterenkonia flava sp. nov. is proposed. The type strain is CAAS 251T (=CCTCC AB 207010T=JCM 14814T).

The purpose of this study was to produce recombinant xylanase in transgenic plants and to test its potential application for pulp bleaching. The xynII xylanase gene from Trichoderma reesei was inserted into the Arabidopsis genome. Many transgenic plants produced biologically active XYNII and accumulated in leaves at level of 1.4-3.2% of total soluble proteins. The bleaching ability of XYNII on Kraft pulp was demonstrated by a reduction in the kappa number and the residual lignin contents. The bleaching efficiency of transgenic plant produced XYNII was similar to commercial xylanase on unbleached Kraft pulp. The effect of xylanase treatment on Kraft pulp was also investigated by SEM. Clear physical change on the pulp fiber surface was observed and was related to the amount xylan removed and microfibrils were visible on the fiber surface. This report demonstrates the potential application of plant produced recombinant xylanase for pulp and paper bleaching.

In a previous cumulative impacts assessments of fish populations along the Waikato River, New Zealand, reduced ovary pigmentation was observed in the common bully (Gobiomorphus cotidianus) with an associated reduction in recruitment in the pulp mill-impacted Kopakorahi Arm of Lake Maraetai compared to an upstream reference site. Carotenoid pigments are typically responsible for the yellow to red colouration of fish eggs and a number of functions for these pigments such as pro-vitamins and antioxidants have been proposed. The purpose of the current study was first to quantify the observed pigment reduction and then to establish possible consequences of carotenoid deficiency. Significantly lower levels of all measured carotenoids were observed in the ovaries of fish from below the mill outfall compared to the reference site. Using in vitro incubations of ovarian follicles, it was also found that these fish had a significantly reduced ability to produce the sex steroids testosterone and estradiol. This study demonstrates changes in ovarian carotenoids associated with pulp mill effluent exposure. Significantly reduced biosynthetic capacity of ovarian follicles demonstrates reproductive endocrine disruption potential associated with exposure to this particular effluent. These results may provide an important link between reproductive dysfunction, oxidative stress and food chain modifications in pulp mill-impacted systems.

Rice straw was used as an alternative raw material to obtain cellulosics pulps. Pulping was done by using classics reagents as soda (with anthraquinone and parabenzoquinone as aditives), potassium hydroxide and Kraft process. The holocellulose, alpha-cellulose and lignin contents of rice straw (viz. 60.7, 41.2 and 21.9 wt%, respectively) are similar to those of some woody raw materials such as pine and eucalyptus, and various non-wood materials including olive tree prunings, wheat straw and sunflower stalks. Pulping tests were conducted by using soda, soda and anthraquinone at 1 wt%, soda and parabenzoquinone at 1 wt%, potassium hydroxide and sodium sulphate (Kraft process) under two different sets of operating conditions, namely: (a) a 10 wt% reagent concentration, 170 degrees C and 60 min; and (b) 15 wt% reagent, 180 degrees C and 90 min. The solid/liquid ratio was 6 in both cases. Paper sheets made from pulp extracted by cooking with soda (15 wt%) and AQ (1 wt%) at 180 degrees C and 90 min pulp exhibit the best drainage index, breaking length, stretch and burst index (viz. 23 degrees SR, 3494 m, 3.34% and 2.51 kN/g, respectively).

The influence of temperature in the hydrothermal treatment of sunflower stalks on the composition of the liquid fraction obtained was examined. The remaining solid fraction was subjected to ethanol pulping in order to obtain pulp that was used to produce paper sheets. The pulp was characterized in terms of yield, kappa index, viscosity, and cellulose, hemicellulose and lignin contents; and the paper sheets in terms of breaking length, stretch, burst index and tear index. Hydrothermal treatment of the raw material at 190 degrees C provided a liquid phase with maximal hemicellulose-derived oligomers and monosaccharide (glucose, xylose and arabinose) contents (26.9 and 4.2 g/L, respectively). Pulping the solid fraction obtained by hydrothermal treatment at 180 degrees C, with 70% ethanol at a liquid/solid ratio of 8:1 at 170 degrees C for 120 min provided pulp with properties on a par with those of soda pulp from the sunflower stalks, namely: 36.3% yield, 69.1% cellulose, 12.6% hemicellulose, 18.2% lignin and 551 ml/g viscosity. Also, paper sheets obtained from the ethanol pulp were similar in breaking length (3.8 km), stretch (1.23%), burst index (1.15 kN/g) and tear index (2.04 m Nm(2)/g) to those provided by soda pulp.

We studied the influence of the operational variables (viz. ethylene-glycol concentrations of 50–70%, temperatures of 155–185 °C, times of 30–90 min and numbers of PFI beating revolutions of 500–1500) on pulp yield and various paper properties (breaking length, stretch, burst index, tear index and brightness) obtained in the ethylene-glycol pulping of vine shoots, cotton stalks, leucaena ( Leucaena leucocephala ) and tagasaste ( Chamaecytisus proliferus ). The fuzzy neural network models used reproduced the experimental results with errors less than 15% and smaller than those provided by second-order polynomial models in all cases. An ethylene-glycol concentration of 65% at 180 °C for 75 min and 1500 PFI beating revolutions were found to provide substantial savings in energy, chemicals and facility investments as a result of operating under milder conditions than the strongest ones studied in this work. Tagasaste was found to be the most suitable raw material among those tested as it provided the paper sheets with the highest breaking length (4644 m), stretch (2.87%), burst index (2.46 kN/g), tear index (0.33 m Nm 2 /g) and brightness (40.92%); its pulp yield was also high (62.88%), which reflects efficient use of this raw material.

In this work, we studied the influence of operational variables in the bleaching of soda pulp of Musa textilis nee (abaca) [viz. temperature (55-85 degrees C), bleaching time (30-150 min) and peracetic acid concentration oven dry pulp (0.5-4.5%)] on the kappa number and viscosity of the bleached pulp, as well as on the breaking length, burst index and brightness of paper sheets made from it. For this purpose, we used a central composite factorial design in order to identify the optimum operating conditions. In this way equations relating the dependent variables to the operational variables of the bleaching process were derived. These equations reproduce the dependent variables with errors less than 12% for all, except the viscosity which was predicted with errors less than 18%. Obtaining bleached pulp with the highest possible viscosity (1519 ml/g), and paper sheets with the maximum possible breaking length (6547 m) and burst index (5.00 kN/g), entails using a temperature of 55 degrees C, a peracetic acid concentration of 4.5% and a bleaching time of 150 min. This provides a brightness of 79.90%, which is only 6.53% lower than the maximum possible value (85.48%).

The potential of a laccase mediator system on the delignification of Eucalyptus globulus kraft pulp was investigated under diverse operating conditions. A series of enzymatic treatments were performed to determine how biodelignification is influenced by pulp consistency, oxygen pressure, reaction time, dose of laccase, and dose of mediator. The mediator used was violuric acid. The results revealed that the extent of biodelignification remains practically constant between 1% and 2.5% of pulp consistency then decreases. However, its effect is less pronounced than the mediator or laccase charge. In fact, an increase in the mediator charge from 1% to 4% leads to an increase in the extent of delignification (fixed laccase charge = 20 IU/g and reaction time 120 min) from 28.4% to 52.2%. On the other hand, at a mediator charge of 4%, the increase of laccase charge from 10 to 40 IU/g has a similar impact on delignification. As for oxygen pressure, the extent of delignification increases with oxygen pressure, from 38.9% to 48.6%, when air and pure oxygen at 4 bar gauge are used. From the standpoint of reaction time, delignification is characterized by a very fast phase followed by a much slower one, leading to a plateau.

To study the possibility of some residues from pulp and paper industry being used as substrates to produce seedlings in containers, three composting experiments were carried out using eucalyptus bark, pine bark and a mixture (60:40, v:v) of pine bark+eucalyptus bark. Biochemical parameters studied were: acid and alkaline phosphatases, lipase (C10), protease, urease, beta-glucosidase and total cellulases. The microbiological populations of total aerobic bacteria, total fungi, actinomycetes, nitrifying bacteria, cellulolytic bacteria and fungi were also evaluated. At the end of the process physicochemical characterization of composts was also performed. Results showed in general that the highest microbiological populations as well as for enzymatic activities occurred during the thermophilic phase (40 degrees C) of the process. On the other hand and according to the physicochemical characteristics of composts pine bark is the most appropriate material to be used in the formulation of substrates to produce plants in containers.

A análise morfométrica das fibras das folhas de Astrocaryum murumuru var. murumuru Mart. revelou que as fibras do pecíolo apresentaram comprimento e espessura da parede superiores às regiões da ráquis e folíolo, com médias variando de 1.266,09 µm a 3.270,56 µm e 5,56 µm a 10,67 µm, respectivamente. Em relação ao índice de Runckel e coeficiente de flexibilidade, as regiões pecíolo e ráquis obtiveram valores considerados favoráveis para sua utilização na indústria papeleira e, o índice de enfeltramento demonstrou que provavelmente as fibras dessas regiões apresentarão uma boa resistência ao rasgo quando submetidas às avaliações físico-mecânicas. Diante dos resultados, a espécie se revela promissora como fonte alternativa de matéria-prima para a produção de papel, sendo necessários, entretanto, estudos de resistências físico-mecânicas a consolidação deste pré-diagnóstico. The analysis morphometric of the leaf fibers of Astrocaryum murumuru var. murumuru Mart. demonstrated that the petiole's fibers had presented length and wall thickness superiors than the rachis and leaflets, with yours means varying of 1.266,09 µm to 3.270,56 µm and 5,56 µm to 10,67 µm, respectively. In relation the index of Runckel and flexibility's coefficient the petiole and rachis had gotten values considered favorable for its use in the paper industry and, the felting index it probably demonstrated that the fibers of these regions will present good crash resistance when submitted to the evaluations physical-mechanical. therefore, the results show the species is a alternative source of raw material to produce paper, but its necessary to do physical-mechanical resistance studies to consolidate this pre diagnostic.

Alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost is an actinomycete that produces industrially important and environmentally safer thermostable cellulase-free xylanase, which is used in the pulp and paper industry as an alternative to the use of toxic chlorinated compounds. Strain CKMX1 was previously characterized by metabolic fingerprinting, whole-cell fatty acids methyl ester analysis, and 16S rDNA and was found to be C. cellulans CKMX1. Crude enzyme (1027.65 U/g DBP) produced by C. cellulans CKMX1, having pH and temperature optima of 8.0 and 60 °C, respectively, in solid state fermentation of apple pomace, was used in the production of bleached wheat straw pulp. Pretreatment with xylanase at a dose of 5 U/g after pulping decreased pulp kappa points by 1.4 as compared with the control. Prebleaching with a xylanase dose of 5 U/g pulp reduced the chlorine charge by 12.5%, increased the final brightness points by approximately 1.42% ISO, and improved the pulp strength properties. Xylanase could be substituted for alkali extraction in C–Ep–D sequence and used for treating chemically bleached pulp, resulting in bleached pulp with higher strength properties. Modification of bleached pulp with 5 U of enzyme/g increased pulp whiteness and breaking length by 1.03% and 60 m, respectively; decreased tear factor of pulp by 7.29%; increased bulk weight by 3.99%, as compared with the original pulp. Reducing sugars and UV-absorbing lignin-derived compound values were considerably higher in xylanase-treated samples. Cellulosimicrobium cellulans CKMX1 has a potential application in the pulp and paper industries.

An alkalistable endoxylanase from Streptomyces cyaneus SN32 was applied in bleaching of wheat straw enriched soda pulp. The xylanase dose of 10 IUg(-1) moisture free pulp exhibited maximum bleach boosting of soda pulp (pH 9.5-10.0) optimally at 65 degrees C after 2 h of reaction time. Pre-treatment of pulp with xylanase and its subsequent treatment with 6% hypochlorite reduced the kappa number by 8.7%, enhanced the brightness index by 3.56% and improved other paper properties such as tear index and burst index. The enzymatically-prebleached pulp when treated with 10% reduced level of hypochlorite (5.4%) gave comparable brightness of resultant hand sheets to the fully bleached pulp (6% hypochlorite).

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.

The aim of this study was to characterize the actinobacterial population present in pulps and boards containing recycled fibres. A total of 107 isolates was identified on the basis of their pigmentation, morphological properties, fatty acid profiles and growth temperature. Of the wet pulp and water sample isolates (n=87), 74.7% belonged to the genus Streptomyces, 17.2% to Nocardiopsis and 8.0% to thermoactinomycetes, whereas all the board sample isolates (n=20) were thermoactinomycetes. The identification of 53 isolates was continued by molecular methods. Partial 16S rDNA sequencing and automated ribotyping divided the Streptomyces isolates (n=31) into 14 different taxa. The most common streptomycetes were the mesophilic S. albidoflavus and moderately thermophilic S. thermocarboxydus. The Nocardiopsis isolates (n=11) belonged to six different taxa, whereas the thermoactinomycetes were mainly members of the species Laceyella sacchari (formerly Thermoactinomyces sacchari). The results indicated the probable presence of one or more new species within each of these genera. Obviously, the drying stage used in the board making processes had eliminated all members of the species Streptomyces and Nocardiopsis present in the wet recycled fibre pulp samples. Only the thermotolerant endospores of L. sacchari were still present in the final products. The potential of automated ribotyping for identifying actinobacteria was indicated, as soon as comprehensive identification libraries became available.

This study investigated the effects of cyanobacteria from pulp-and-paper waste-treatment systems on biological toxicity removal and biodegradation of certain wastewater contaminants. In field and batch studies, using the Microtox®assay, cyanobacterial biomass and final wastewater toxicity were significantly correlated. In softwood-based wastewater, a decrease in toxicity was negatively correlated with cyanobacterial biomass, but the correlation was positive in hardwood-based wastewater. In the softwood-based wastewater, toxicity remained higher in the light than it was in the dark, whereas in hardwood-based wastewater, toxicity was lower in the light than it was in the dark. All of these results were light-dependent, suggesting that the photosynthetic growth of cyanobacteria is required to induce significant effects. When grown in mixed cultures with bacterial degraders, cyanobacteria from pulp-and-paper waste-treatment systems generally impeded the biodegradation of the wastewater contaminants phenol and dichloroacetate (DCA). However, there was one case where the cyanobacterium Phormidium insigne improved the bacterial degradation of DCA. Doubling inorganic nutrient concentrations did not improve phenol or DCA biodegradation in the majority of cases, indicating that nutrient competition is not a major factor. These data suggest that cyanobacteria play an important role during the biological treatment of contaminants, and, hence, toxicity removal in pulp-and-paper waste-treatment systems.Key words: cyanobacteria, heterotrophic bacteria, biodegradation, pulp and paper waste-treatment, wastewater toxicity.

A comparative study on TCF (totally chlorine-free) bleachability of organosolv pulps from the annual fibre crop Arundo donax L. (giant reed) was carried out using a simple three-stage peroxide bleaching sequence without oxygen pre-bleaching. ASAM (alkali-sulfite-anthraquinone-methanol), Organocell (alkali-anthraquinone-methanol) and ethanol-soda organosolv pulps were bleached and compared with kraft pulp, as a reference. The final brightness of 76-78% ISO was attained for all tested pulps. The chemical charge required to reach this level of brightness varied for different pulps (despite the equal initial content of the residual lignin) and directly related to starting brightness values. No direct correlation between brightness improvement and lignin removal during bleaching was found, indicating the influence of the specific pulp properties introduced by pulping process on bleaching chemistry. The general higher bleaching response of organosolv pulps from A. donax was noted in comparison with kraft.

Alkaline peroxide mechanical pulping (APMP) of wheat straw with enzyme treatment was studied. Instead of direct enzyme pretreatment on wheat straw, an alternative treatment method was used, in which coarse pulps from refiner defibrated wheat straw rather than wheat straw were pretreated with a crude enzyme containing mainly xylanase, then impregnated with alkaline H2O2 solution and further refined. The optimum conditions of enzyme treatment were xylanase dosage of 10-15 IU/g of oven-dried wheat straw, 90 min, 50-60 degrees C, pulp consistency of 5-10%, and initial pH of 5.0, and those for chemical impregnation were 6% NaOH, 70-80 degrees C, 60-90 min, and 4 to 5% H2O2. Enzyme treatment improved pulpability of wheat straw by the APMP process, and final pulp quality such as brightness, breaking length, and burst index of pulp. Pulp from the APMP process with enzyme treatment could be bleached to a brightness of 70.5% ISO by two-stage H2O2 bleaching sequence with only 4% H2O2, and breaking length of the bleach pulp reached 4470 m