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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.

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.

Bacterial strains were isolated from spruce wood chips and their ability to reduce the content of wood extractives was studied. Strains were screened by cultivation on liquid media containing wood extractives as the major nutrient. Some bacterial species could decrease remarkably the amount of extractives in the liquid media and reduced the amount of triglycerides, steryl esters and total extractives by 100, 20 and 39%, respectively. Spruce wood chips were treated in controlled conditions with selected bacteria to test their effects on the chips. All the bacteria grew well on wood chips. The effect of bacterial metabolism on wood extractives was significant. Bacterial treatments reduced the amount of lipophilic extractives by 16-38% in 1 week of treatment and up to 67% in 2 weeks. The most efficient strain removed 90, 66 and 50% of triglycerides, steryl esters and resin acids, respectively, in 2 weeks. These results indicate that bacteria may be promising agents for the removal of extractives for improved pulping and papermaking processes.

Agar substrates forin vitroculture are well adapted to plant micropropagation, but not to plant rooting and acclimatization. Conversely, paper-pulp-based substrates appear as potentially well adapted forin vitroculture and functional root production. To reinforce this hypothesis, this study comparesin vitrodevelopment of nemesia on several substrates. Strong differences between nemesia roots growing in agar or in paper-pulp substrates were evidenced through scanning electron microscopy. Roots developed in agar have shorter hairs, larger rhizodermal cells, and less organized root caps than those growing on paper pulp. In conclusion, it should be noted that in this study,in vitromicroporous substrates such as paper pulp lead to the production of similar root hairs to those found in greenhouse peat substrates. Consequently, if agar could be used for micropropagation, rooting, and plant acclimatization, enhancement could be achieved if rooting stage was performed on micro-porous substrates such as paper pulp.

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

International audience; Pycnoporus cinnabarinus laccase was fused to the C-terminal linker and carbohydrate binding module (CBM) of Aspergillus niger cellobiohydrolase B (CBHB). The chimeric enzyme of molecular mass 100 kDa was successfully produced in A. niger. Laccase-CBM was further purified to determine its main biochemical properties. The Michaelis-Menten constant and pH activity profile were not modified, but the chimeric enzyme was less thermostable than either the P. cinnabarinus laccase or the recombinant laccase produced in the same strain. Laccase-CBM was able to bind to a cellulosic substrate and, to a greater extent, to softwood kraft pulp. Binding to the pulp was shown to be mainly time and temperature-dependent. Laccase-CBM was further investigated for its softwood kraft pulp biobleaching potential and compared with the P. cinnabarinus laccase. Addition of a CBM was shown to greatly improve the delignification capabilities of the laccase in the presence of 1-hydroxybenzotriazole (HBT). In addition, ClO2 reduction using 5 U of chimeric enzyme per gram of pulp was almost double than that observed using 20 U of P. cinnabarinus laccase per gram of pulp. We demonstrated that conferring a carbohydrate binding capability to the laccase could significantly enhance its biobleaching properties

Lipophilic low molar-mass constituents in wood chips for the paper industry result in low quality pulp, pitch deposition, and effluent toxicity. New biotechnological solutions such as fungal pre-treatment of wood chips can reduce pitch problems. This laboratory-scale study focuses on the potential and limitations of a fungal bio-treatment of Norway spruce chips with the white-rot fungus Trametes versicolor. Different fungal treatment conditions were compared. A 4-week fungal treatment reduced the concentration of resin acids and triglycerides by 40% and 100%, respectively, but neither lowered the energy requirements of the TMP process nor significantly affected the morphological fiber characteristics and the physical pulp properties. The pre-treatment led to slightly poorer optical properties. The Trametes versicolor fungal treatment contributed to a less toxic effluent and improved the biodegradability. A treatment of 2-3 weeks appears optimal.

Studies were carried out in vitro on a cellulose based agar at two water activities ( a w ,0.975,0.995) and on three types of paper at two relative humidities (75, 100% RH) for the potential for differentiation of contamination and colonisation by Aspergillus terreus , A. holandicus and Eurotium chevalieri . In vitro studies showed that conducting polymer sensor array gave different responses to each of these species when grown on cellulose agar at both a w levels. Discriminant function analyses of the data showed differentiation of the controls from the spoilage fungi. Cluster analysis gave a significant ( P =0.05) separation of the control and each spoilage fungus. In situ studies on three types of paper showed that using natural substrates the volatile patterns produced by each of these fungi was different from each other and from the control. The results obtained were better at the higher humidity. The three paper types could be successfully differentiated into clusters. For a single paper type, differentiation of controls from spoilage fungal treatments was better at the higher humidity. This study has shown that this technology has potential for the early detection of fungal contamination in library materials and archives for the improved protection of cultural heritage.

International audience; The agronomic and pulping performance of transgenic trees with altered lignin has been evaluated in duplicated, long-term field trials. Poplars expressing cinnamyl alcohol dehydrogenase (CAD) or caffeate/5-hydroxy-ferulate O-methyltransferase (COMT) antisense transgenes were grown for four years at two sites, in France and England. The trees remained healthy throughout the trial. Growth indicators and interactions with insects were normal. No changes in soil microbial communities were detected beneath the transgenic trees. The expected modifications to lignin were maintained in the transgenics over four years, at both sites. Kraft pulping of tree trunks showed that the reduced-CAD lines had improved characteristics, allowing easier delignification, using smaller amounts of chemicals, while yielding more high-quality pulp. This work highlights the potential of engineering wood quality for more environmentally benign papermaking without interfering with tree growth or fitness.

A unicellular blue-green alga, Agmenellum quadruplicatum , and a filamentous blue-green alga, Lyngbya lagerheimíi , were grown heterotrophically in dim light with glucose as major source of carbon and possibly energy. The dim-light conditions did not support autotrophic growth. The two blue-green algae appeared to have the same metabolic block, namely an incomplete tricarboxylic acid cycle, as has been found in other obligately phototrophic blue-green algae. Under dim-light conditions, glucose made a greater contribution to cell constituents (amino acids) of A. quadruplicatum and L. lagerheimii than under high-light conditions.

Microbial penetration of sterile packs was studied by using double-wrap (two layers each) muslin, single-wrap (two layers) muslin inner covering with single-wrap (one layer) two-way crepe paper outer covering, and single-wrap (two layers) muslin inner covering with single-layer BAR-BAC wrappers to wrap 20 gauze sponges (2 by 2 in.). These packs were stored on open shelves of a central sterile supply department of a hospital and processed for sterility at weekly intervals. Microorganisms penetrated the double-wrap muslin as early as 28 days, the single-wrap muslin and single-wrap two-way crepe paper combination in 77 days, and the single-wrap muslin and single-layer BAR-BAC combination in 63 days.