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Background For microorganisms on a paper surface, the lack of water is one of the most important stress factors. A strain of Bacillus megaterium FDU301 was isolated from plaques on a paper surface using culture medium with polyethylene glycol 200 (PEG200) to simulate an arid condition. Global transcriptomic analysis of B. megaterium FDU301 grown under normal and simulated arid conditions was performed via RNA-seq technology to identify genes involved in arid stress adaptation. Results The transcriptome of B. megaterium FDU301 grown in LB medium under arid (15% PEG200 (w/w)) and normal conditions were compared. A total of 2941 genes were differentially expressed, including 1422 genes upregulated and 1519 genes downregulated under arid conditions. Oxidative stress-responsive regulatory genes perR, fur, and tipA were significantly upregulated, along with DNA protecting protein (dps), and catalase (katE). Genes related to Fe2+ uptake (feoB), sporulation stage II (spoIIB, spoIIE, spoIIGA), small acid-soluble spore protein (sspD), and biosynthesis of compatible solute ectoine (ectB, ectA) were also highly expressed to various degrees. Oxidative phosphorylation-related genes (atpB, atpE, atpF, atpH, atpA, atpG, atpD, atpC) and glycolysis-related genes (pgk, tpiA, frmA) were significantly downregulated. Conclusion This is the first report about transcriptomic analysis of a B. megaterium to explore the mechanism of arid resistance. Major changes in transcription were seen in the arid condition simulated by PEG200 (15%), with the most important one being genes related to oxidative stress. The results showed a complex mechanism for the bacteria to adapt to arid stress.

A goal of our study was to find an alternative to nano-silver-based antimicrobial materials which would contain active silver immobilized in a solid matrix that prevents its migration into the surrounding environment. In this study, we investigated whether silver cations dispersed in an atomic form and trapped in an ion-exchanged zeolite show comparable antimicrobial activity to silver nanoparticles (NPs). The biocidal active material was prepared from the sodium form of faujasite type zeolite in two steps: (1) exchange with silver cations, (2) removal of the external silver oxide NPs by elution with Na2EDTA solution. The modified biocidal zeolite was then added to paper pulp to obtain sheets. The zeolite paper samples and reference samples containing silver NPs were tested in terms of biocidal activity against an array of fungi and bacteria strains, including Escherichia coli, Serratia marcescens, Bacillus subtilis, Bacillus megaterium, Trichoderma viride, Chaetomium globosum, Aspergillus niger, Cladosporium cladosporioides, and Mortierella alpina. The paper with the modified faujasite additive showed higher or similar antibacterial and antifungal activities towards the majority of tested microbes in comparison with the silver NP-filled paper. A reverse effect was observed for the Mortierella alpina strain.

Liquid packaging boards and blanks were examined for microbial contaminants. A total of 218 strains were identified and representatives of the most frequent species were characterized for their potential for food spoilage. Contaminants found were aerobic spore-forming bacteria, mostly Bacillus megaterium, B. licheniformis, B. cereus group, B. pumilus, Paenibacillus macerans, P. polymyxa, P. pabuli and B. flexus. Production of amylolytic, proteolytic, lipolytic and phospholipolytic enzymes was common. Approximately 50% of the B. cereus group strains were positive in the diarrhoeal enterotoxin immunoassay test or in the enterotoxin reversed passive latex agglutination test. Strains capable of growth at 6 degrees C were found among B. cereus group, P. pabuli, P. validus, B. megaterium and P. polymyxa. All b. licheniformis strains grew at 55 degrees C. The spores of B. licheniformis were most resistant to hydrogen peroxide. The B. cereus group strains were recognizable by fatty acid components not present in any of the other paperboard strains, 11-methyldodecanoic acid (13:0 iso) and trans-9-hexadecenoic acid (16:1 omega 7 trans), each contributing 7% or more to the total cellular fatty acids.

Aims: To survey paperboard products from 17 US mills for bacterial populations and for bacteria potentially harmful to human health. Methods and Results: Culturable aerobic bacteria were isolated from paperboard products using selective and nonselective medium. Resulting colonies from samples from three regions of the United States were identified using fatty acid methyl ester analysis. Percentages of bacteria species found were Bacillus megaterium (47), Bacillus licheniformis (15), Bacillus pumilus (12), Paenibacillus macerans (5), Paenibacillus pabuli (3), Bacillus subtilis (2), Bacillus cereus (2), Bacillus coagulans (1), Bacillus circulans (1), Bacillus brevis (1), Bacillus thuringiensis (1), Paenibacillus polymyxa (1), Cellulomonas turbata (1), Cellulomonas flavigena (1), unidentified Bacillus sp. (3) and unidentified bacteria (1). Conclusions: Recycled paperboard contained high populations of bacteria, and a positive correlation was found between recycle content and bacterial populations. Escherichia coli, Salmonella, Shigella or confirmed coliform bacteria were not found in any product. Significance and Impact of the Study: Populations of bacteria did not differ significantly from original counts over a 4-month period of dry storage, indicating that bacteria persist in paperboard over long periods and may re-enter the recycling process. The predominance of heat-tolerant endospore-forming bacteria explains the high bacteria counts found in paperboard made from recycled materials.

Xylanase production from B. megaterium was enhanced using solid state fermentation with respect to the use of solid substrate, moistening solution, moisture content, inoculum, sugars, soyabean meal, amino acids, and extraction with surfactant. An increase of approximately 423-fold in xylanase production and complete suppression of CMCase production was achieved over submerged liquid fermentation. Biobleaching using this cellulase-free xylanase, 8 U/g of oven dried pulp of 10% consistency, showed 8.12% and 1.16% increase in brightness and viscosity, 13.67% decrease in kappa number, and 31% decrease in chlorine consumption at the CD stage.

Aims: To identify aerobic mesophilic bacteria isolated from coreboard, kitchen roll paper and food packaging boards containing recycled fibres and to create a rapid fingerprint‐based database for their identification.Methods and Results: A total of 197 isolates and 20 relevant type strains were characterized by automated ribotyping and as far as possible identified by the similarities of their riboprints to the relevant type strains. One strain from each unidentified ribotype, a total of 87 strains, was subjected to partial 16S rDNA sequencing and in most cases also to fatty acid analysis and physiological tests. From the isolates 113 and seven different ribotypes were generated belonging to the genera Bacillus and Paenibacillus, respectively. The dominating species, or closest related to them, were B. simplex (22·8% of isolates), B. licheniformis (18·3%) and B. amyloliquefaciens (12·7%); 5·1% of the isolates were identified as B. cereus, a potential food‐borne pathogen. In particular, this species was present in one food packaging board (26·3% of isolates). Based on these results, 40·1% of the isolates and 45·0% of ribotypes were so different from the relevant type strains that they may represent novel species.Conclusions:All isolates were aerobic spore‐formers, indicating that all non‐spore‐formers were eliminated during the drying stage of the processes. Although many isolates could be affiliated to described species of Bacillus or Paenibacillus, a significant proportion of the isolates could not be identified unambiguously as members of a described species.Significance and Impact of the Study: A RiboPrint® identification database, composed of 120 composite patters, was established for bacteria originating from the pulp and paper industry. Considering the discrimination power of ribotyping, this database will be extremely useful in future for the reliable and rapid identification of bacteria isolated from pulp and paper industrial sources.