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Desulfovibrionales-related bacteria in a paper mill environment as detected with molecular techniques and culture.

The aim of the present study was to evaluate the suitability of a nested PCR-DGGE (denaturing gradient gel electrophoresis) method for the detection of Desulfovibrionales-related sulfate-reducing bacteria (SRB) from paper mill samples. The samples were also analyzed with culturing. SRB cause/enhance industrial problems, namely creation of foul-smelling gases (hydrogen sulfide) and biological corrosion, and so far there has not been a simple method to study these bacteria in paper mill laboratories. In our study, culturing was able to detect Desulfovibrionales-related bacteria from two different white waters, two different brokes, pulp, clay, and slime. Out of the isolated Desulfovibrionales, 23 enrichment cultures were further characterized with Desulfovibrionales-selective PCR-DGGE. An identical Desulfovibrio species sequence was found from paper machine I (broke I, slime, and pulp) and from paper machine II (broke II and white water II), suggesting an in-house contamination with the same strain. Desulfovibrionales-selective PCR-DGGE was also performed from DNA templates extracted directly from the paper mill samples. The DGGE profiles derived from the samples without prior enrichment were more diverse and the sequenced amplicons proved to belong to the Desulfovibrionales order. Moreover, molecular techniques were able to detect Desulfovibrionales-related bacteria from calcium carbonate samples whereas culture did not. Altogether, the nested PCR-DGGE method used in this study was suitable for the detection of Desulfovibrionales-related SRB directly from different paper mill samples and it could be used for the rapid identification of SRB-contaminated industrial sites and, when combined with sequencing, for tracing of the contamination routes.

Bacterial Typing Techniques↗

Diversity of bacteria contaminating paper machines.

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.

Bacteria↗

Seasonal bacteriological analysis of Gola river water contaminated with pulp paper mill waste in Uttaranchal, India.

The seasonal physico-chemical and microbial quality of Gola river water has been analyzed after confluence of pulp paper mill waste. The study revealed that it has enhanced 20-30 times pollution load of BOD, COD, TDS, TSS, sulphate, chloride, sodium, nitrate, potassium, lignin and phenol after mixing of pulp paper mill waste with river water in all season. Further, it induced the bacterial growth by increasing most probable number value of E. coli was 1.57 x 10(4), 1.6 x 10(4), 1.37 x 10(4) and SPC count was 1.68 x 10(4), 1.64 x 10(4), 1.67 x 10(4)/100 ml during summer, monsoon, winter respectively. While the most probable number value in river water before mixing of pulp paper mill waste was 1.4 x 10(2), 1.82 x 10(2), 1.5 x 10(2) and SPC count was 2.8 x 10(3), 2.89 x 10(3), 2.78 x 10(3)/100 ml during summer, monsoon and winter respectively. This indicated from 88-114 fold increase in most probable number value of E. coli and 56.55-60.0 times increase in SPC count of river water after mixing of effluent in summer, monsoon and winter. Moreover, the most probable number value in effluent itself before mixing was 3.4 x 10(2), 3.3 x 10(2), 2.8 x 10(2) and SPC count was 6 x 10(4), 6.5 x 10(4), 6 x 10(4)/100 ml during summer, monsoon, winter, respectively. Furthermore, it was revealed that the seasonal variation also regulated the bacterial population dynamics as per the physico-chemical quality, in which E. coli was found highest at the rate of (5.9 x 10(4)), E. aerogenes (5.3 x 10(4)), P. aeruginosa (1.3 x 10(4)), S. aureus (3.2 x 10(3)), K. pneumoniae (2.6 x 10(4)), Enteritidis (1.1 x 10(4)) on monsoon season and V. cholerae (7.4 x 10(2)), V. vulnificus (9.2 x 10(2))/100 ml in river water when contaminated with pulp paper mill waste in monsoon season. Thus, the monsoon season showed presence of FC and TC indicated the thermo-tolerant and disease causing group of bacterial population in effluent and its sequence was observed as monsoon>summer>winter. This indicated the growth of many pathogenic and non-pathogenic bacteria for health hazards with contamination of pulp paper waste in aquatic ecosystem within the vicinity of pulp paper mill industry.

Chlorides↗

Modelling the absorption and desorption of cadmium on paper pulp using kinetic approaches.

The presence of toxic metals on paper pulp and the migration of these metals to food from the food package is receiving significant attention. The final exposure levels for consumers depend on two main processes. First the potential of metals to bind paper pulp during manufacture. Second, the metal potential to migrate from paper to food during storage and use. Binding and unbinding processes are modelled for cadmium kinetics through kinetic approaches. The cadmium concentration in paper pulp is estimated from the cadmium concentration in the water-pulp liquor during manufacture, the temperature, and contact time. Two food simulants have been employed for desorption studies, water and acetic solution (3%, w/w). As expected, under acidic conditions desorption is total and rapid (close to 100% desorption reached in a few minutes). However, the desorption of cadmium into the neutral food simulant depends on the initial cadmium concentration in the paper pulp, temperature and contact time. Surface response curves for each combination are presented. Temperature did not affect cadmium binding, but played a significant role for the desorption processes into the neutral food simulant. The proposed equations offer a good fitting of the experimental values, with p < 0.001 and residuals within a factor of 3 for over 99% of the measured data. These models allow estimations of the expected exposure levels in consumers, on the basis of manufacture and use conditions. Linking the expected exposure with toxicity thresholds, such as the admissible daily intake (ADI), would allow a scientific assessment of the maximum acceptable cadmium levels in water-pulp liquors during manufacture and in the final paper pulp on the basic of the use patterns of each food packaging material.

Adsorption↗

Proximity to pulp and paper mills and wheezing symptoms among adolescents in North Carolina.

Using data from the North Carolina School Asthma Survey about the respiratory health of 64,432 adolescents attending public schools in North Carolina and data provided by school employees about the environmental health conditions of the school buildings, we assessed the prevalence of daytime wheezing during the past year among students estimated to be exposed to air pollutants arising from pulp and paper mills located near the schools. Of the schools the students attended, 14% (37/266) were located within 30 miles of one or more pulp and paper mills and odor from the mills was identified by survey respondents for 9 of the 266 schools. The prevalences of daytime wheezing in smokers and nonsmokers with household cigarette smoke exposure were elevated among students attending schools located within 30 miles of a pulp and paper mill, compared to the prevalence among students attending schools located beyond 30 miles (10 miles, prevalence ratio (PR): 1.21, 95% confidence interval (95% CI): 0.99, 1.43; 10-30 miles, PR: 1.06, 95% CI: 0.96, 1.16) and among students attending schools with noticeable odor from a pulp and paper mill (PR: 1.12, 95% CI: 0.93, 1.31). These results indicate a possible association between paper mill location and wheezing symptoms among adolescents and suggest that community-based exposure to pulp and paper mill emissions may have a greater impact on smokers and individuals exposed to cigarette smoke in the home than on nonsmokers without such household exposure.

Adolescent↗

A paper screening test to assess genetic taste sensitivity to 6-n-propylthiouracil.

The purpose of this study was to develop a quantifiable method to fix 6-n-propylthiouracil (PROP) onto filter paper disks and to test the validity of the method relative to the three-solution test, previously developed in this laboratory. Filter paper disks were impregnated with 50 mmol/l PROP or 1.0 mol/l NaCl then dried. The concentration of PROP per disk was determined to be 0.280 mg+/-2.2% (CV) by ethanol extraction. Subjects were studied in two groups (n=62 per group). All subjects were initially classified by the three-solution test and then were independently classified by the paper disk test. In the paper disk test, group means for PROP (+/-95% confidence interval) were calculated and used to establish numerical cutoff scores for taster group classification. Cutoff scores for Group 1 were used to classify subjects in Group 2, verifying that the cutoffs were applicable to another subject group. The percentage of nontasters, medium tasters, and supertasters identified by paper disk was 27%, 42%, and 31%, respectively, which is consistent with expected frequencies in the population. For Group 1, the agreement between the two classification methods ranged from 86% to 94% across taster groups. For Group 2, the agreement ranged from 83% to 100%. The contingency coefficient (P) of the degree of association between the two classification methods was high (P=0.77 and P=0.74 for Groups 1 and 2, respectively; P< or =.001). These results demonstrate that the paper disk method is a reliable screening tool for assessing sensitivity to PROP that has numerous applications in basic and applied research.

Adult↗

Sequential (anaerobic/aerobic) biological treatment of Dalaman SEKA pulp and paper industry effluent.

In the pulp and paper industry, lignin and other color compounds are removed by chemical agents in bleaching process. Use of chlorine-based agents results in production of degradation products which include various chloro-organic derivatives. Since these new compounds are highly chlorinated, they cause a problem in the treatment of pulp and paper industry wastewaters. Chemical precipitation, lagooning, activated sludge, and anaerobic treatment are the processes used for treating pulp and paper effluents. Furthermore, a combination of these processes is also applicable. In this study, the effluent of Dalaman SEKA Pulp and Paper Industry was examined for its toxic effects on anaerobic microorganisms by anaerobic toxicity assay. Additionally, this wastewater was applied to a sequential biotreatment process consisting of an upflow anaerobic sludge blanket as the anaerobic stage and a once-through completely mixed stirred tank as the aerobic stage. Results indicated that: (1) Dalaman SEKA Pulp and Paper Industry wastewater exerted no inhibitory effects on the anaerobic cultures under the studied conditions, and (2) application of a sequential biological (anaerobic/aerobic) system to treat the Dalaman SEKA Pulp and Paper Industry wastewater resulted in approximately 91% COD and 58% AOX removals at a HRT of 5 and 6.54 h for anaerobic and aerobic, respectively.

Bacteria, Aerobic↗

Improvement of the physical properties of reprocessed paper by using biological treatment with modified cellulase.

A primary need for waste paper reprocessing is to preserve optical properties and the physical strength of the paper fibers. In this study, modified cellulase with copolymer, polyethylene oxide (PEO) derivatives and maleic anhydride (MA) was applied to the reprocessing of mixed office waste (MOW). Modified cellulase was prepared by a chemical reaction between amino groups of the cellulase and the MA functional groups of the copolymer. In MOW reprocessing, modified cellulase improved several physical properties of the paper including freeness, optical properties, and physical strength compared to the conventional process. Even though native cellulase improved the physical properties, paper treated with modified cellulase exhibited an increase in physical properties such as tensile strength and internal bond over those of unmodified cellulase. From these results, modified cellulase method is a new biological treatment that will save pulp resources, which are added to waste paper reprocessing to maintain the strength of paper.

Cellulase↗

Effect of cellulose/hemicellulose and lignin on the bioavailability of toluene sorbed to waste paper.

Paper constitutes about 38% of municipal solid waste, much of which is disposed of in landfills. Sorption to such lignocellulosic materials may limit the bioavailability of organic contaminants in landfills. The objective of this study was to identify the effect of individual biopolymers in paper on toluene sorption and bioavailability by subjecting fresh and anaerobically degraded office paper and newsprint to enzymatic hydrolysis and acid hydrolysis. Enzymatic degradation of cellulose and hemicellulose had no effect on toluene bioavailability. In contrast, acid-insoluble lignin controlled toluene sorption and bioavailability for both fresh and degraded newsprint. Acid-insoluble lignin could explain only 54% of the toluene sorption capacity of degraded office paper however, suggesting that crude protein and/or lipophilic organic matter were also important sorbent phases. Toluene sorbed to degraded office paper was also less bioavailable than toluene sorbed to an equivalent mass of lignin extracted from this sorbent. The latter result suggests that a fraction of toluene sorbed to degraded office paper may have been sequestered by lipophilic organic matter. The sorption and bioavailability data indicate that the preferential decomposition of cellulose and hemicellulose relative to lignin in landfills should not decrease the overall toluene sorption capacity of paperwaste or increase the bioavailability of sorbed toluene.

Adsorption↗

Microbial communities of printing paper machines.

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

Bacteria↗

Neurocysticercosis: validity of ELISA after storage of whole blood and cerebrospinal fluid on paper.

Cysticercosis is an infestation of Cysticercus cellulosae. When it occurs in the brain, chronic neurological complications can ensue, most commonly seizures. Neurocysticercosis is usually diagnosed by neuroimaging, a technique not available in most endemic countries. Hence immunological tests are valuable for diagnosis and epidemiological surveys. We evaluated the suitability of paper for storing blood and cerebrospinal fluid (CSF) until subsequent testing by enzyme-linked immunosorbent assay (ELISA), by testing whole blood samples on filter paper from 305 patients and CSF samples from 117 patients stored on ordinary white typing paper and on filter paper. Optimal preservation of biological samples is achieved when whole blood is stored on filter paper, CSF on white paper, and when samples are frozen within 1 week after collection. Our results could improve diagnostic capabilities and facilitate epidemiological surveys in endemic countries where immunodiagnostic tests cannot be rapidly performed because of inadequate laboratory infrastructure.

Blood↗

Migration of dehydroabietic and abietic acids from paper and paperboard food packaging into food-simulating solvents and Tenax TA.

An investigation was undertaken to establish the concentration in paper products of dehydroabietic (DHA) and abietic (AA) resin acids, present in rosin, which are major toxicants of pulp- and paper-mill effluent. Their migration was studied from paper and paperboard products into various food-simulating solvents and the substitute fatty food simulant Tenax TA (modified polyphenylene oxide). DHA and AA were detected in five of 10 virgin paper products and in all 10 recycled paperboard products for food-contact use at concentrations of 14-500 and 110-1200 microg/g, respectively. In virgin paper products, the highest migration was into 95% ethanol or heptane, with negligible or no migration into other solvents. In recycled paperboard products, migration was highest into 95% ethanol, but was also observed into 20% ethanol, water and heptane. Migration to Tenax TA was also observed and the migration level increased with time. The maximum migration levels of DHA and AA into food simulants were 0.853 and 3.14 microg/g, respectively. The results suggest that, in the worst case, the daily intake of DHA and AA from paper and paperboard products was 50 times lower than the tolerable daily intake of rosin.

Abietanes↗

Inter-test comparison between filter paper absorbed blood eluate and serum for malaria serology by enzyme immunoassay: an operational feasibility.

Antimalarial IgG and IgM were detected by enzyme immunoassay in finger-stick blood samples collected in capillary tubes and also spotted onto Whatman filter paper. Assay was performed in 92 blood samples obtained from 53 falciparum malaria patients, representing 23 fever cases (malaria negative) and 16 healthy individuals. A simple indirect ELISA was done using Plasmodium falciparum lysate and MSP1(19) peptide as antigens. Total IgG and IgM contents were also estimated in individual sera and filter paper eluate by single radial immunodiffusion (SRID). Assay results of both serum and filter paper eluates were compared. The sensitivity and specificity of the assays for IgG measurement were comparable between serum and filter paper eluates (P < 0.001), whereas, in case of IgM, detection level was poor in filter paper eluates as observed by ELISA and SRID. The filter paper eluates may serve the purpose of antigen-specific IgG detection in seroepidemiological surveys.

Analysis of Variance↗

Efficiency of hand drying for removing bacteria from washed hands: comparison of paper towel drying with warm air drying.

OBJECTIVE: To evaluate warm air and paper towel drying for removing bacteria from washed hands. METHODS: After hands were washed with non-antibacterial soap, they were dried using warm air with and without ultraviolet light, while being rubbed or held stationary, or paper towels. Each method was performed as a randomized trial using 30 hands. RESULTS: Log colony-forming units (CFU) on palms and fingers increased significantly when hands were dried with warm air while being rubbed for 15 seconds (P < .001), and many bacteria remained at 30 seconds without ultraviolet light (P < .001) Holding hands stationary while drying significantly decreased log CFU on palms, fingers, and fingertips (P < .01 or < .001). Few CFU were detected on palms and fingers dried with ultraviolet light. Although log CFU of palms and fingers did not decrease after drying with three sheets of paper towel, those of fingertips decreased significantly (P < .001). For palms and fingers, log reductions were greater with warm air drying while holding hands stationary, paper towels, and warm air drying while rubbing hands. For fingertips, the log reduction was often greater with paper towels than with warm air. CONCLUSIONS: Holding hands stationary and not rubbing them was desirable for removing bacteria. Ultraviolet light reinforced the removal of bacteria during warm air drying. Paper towels were useful for removing bacteria from fingertips but not palms and fingers.

Adult↗

Equivalence of litmus paper and intragastric pH probes for intragastric pH monitoring in the intensive care unit.

OBJECTIVE: To compare the accuracy of litmus paper-determined gastric pH to a nasogastric graphite antimony pH probe. DESIGN: A prospective clinical trial of gastric pH determination in patients enrolled in a study of histamine-2-receptor (H2) antagonists. SETTING: The medical intensive care unit (ICU) of a 450-bed county hospital. PATIENTS: Critically ill ICU patients requiring stress ulcer prophylaxis. INTERVENTIONS: Using a crossover design, the patients were randomized to initially receive an H2 antagonist by continuous infusion or intravenous bolus, and subsequently were crossed over to the other limb of the study. MEASUREMENTS AND MAIN RESULTS: Gastric pH was determined using pH-sensitive litmus paper at the initiation of each limb of the study and at 1, 2, 4, and 8 hrs after the initiation of H2 receptor antagonist therapy. In addition, gastric pH was continuously determined over the same time period utilizing a graphite antimony pH probe. Gastric pH measurements determined with litmus paper and intragastric pH probes demonstrated an excellent correlation (r2 = .93, p < .001). McNemar's test of correlated proportions could not demonstrate a significant difference between the two monitoring methods (chi-square = 0.5, p > .47), and the kappa statistic (0.95, p < .001) demonstrated excellent concordance. Bias measurement was 0.01 (95% confidence interval = -0.155 to 0.176). CONCLUSIONS: Measurement of intragastric pH, using pH-sensitive litmus paper, is both sensitive and specific when utilizing a graphite antimony nasogastric pH probe as a reference standard. Litmus paper-determined gastric pH testing is both easy to perform and inexpensive. Therefore, based on the current data, we believe this technique (i.e., litmus paper determined gastric pH testing) to be the method of choice for determination of intragastric pH in patients at risk for stress gastric ulcers in the medical ICU.

Adult↗

Hodgkin's disease, pharyngeal cancer, and soft tissue sarcomas in Danish paper mill workers.

Several studies have indicated an increased cancer risk among paper mill workers, based on rather few cancer cases. The authors studied cancer incidence in a large historical cohort of 14,362 Danish paper mill workers, employed at any time between 1943 and 1990, and followed up until December 31, 1993. Men had an increased risk of pharyngeal cancer (15 observed (obs), standardized incidence ratio (SIR) 1.99, 95% confidence interval (CI) 1.11-3.29). With two additional cases among women, 11 of 17 cases were located in the tonsils. Men also had an increased risk of Hodgkin's disease (18 obs, SIR 2.01, 95% CI 1.19-3.18). Women had an increased risk of soft tissue sarcomas (nine obs, SIR 2.33, 95% CI 1.06-4.43), with a high risk among female paper sorters (eight obs, SIR 3.98, 95% CI 1.71-7.84). The study thus supports some studies' observations of an increased risk of Hodgkin's disease in paper mill workers. The detailed reporting of cancer cases in paper mill workers have furthermore revealed a risk of pharyngeal cancer together with a risk of soft tissue sarcomas in paper sorters.

Adult↗

Evaluation and comparison of the electrostatic dust print lifter and the electrostatic detection apparatus on the development of footwear impressions on paper.

The Electrostatic Dust Print Lifter (EDPL) and the Electrostatic Detection Apparatus(2) (ESDA(2)) were compared to determine if both processes could be used to develop footwear impressions of the same or similar quality and in what order they should be used to develop the highest quality footwear impression. The sensitivity of each technique was also evaluated. The quality of the footwear impressions developed was determined by comparing 25 individual characteristics present on the known shoe to the footwear impressions developed using each technique. The footwear impressions were made by stepping on paper placed over several different surfaces, which included: linoleum, industrial Berber carpet, nylon carpet placed over a (3/8)-in. pad, ceramic tile, cardboard, 1-in. foam, 4-in. foam, cement, asphalt, grass, and mulch. Each of the papers placed on these surfaces was developed using the EDPL before the ESDA(2) and vice versa. The sensitivity test for the ESDA(2) was conducted by processing 10 sheets of stacked paper that were stepped on with the known shoe, beginning with the top sheet. The sensitivity test for the EDPL was conducted by processing 10 sheets of paper stepped on with the known shoe in succession. This study determined the footwear impressions developed using the EDPL were of better comparative value than impressions developed with the ESDA(2). On average, 72.4% of the individual characteristics from the known impression were identified on images developed when the EDPL was used first compared with an average of 38.9% when the ESDA(2) was used first. Therefore, if only one technique is used, the EDPL should be chosen. The sensitivity test determined the ESDA(2) develops high-quality footwear impressions on only the top sheet of paper. No footwear impressions were developed on any sheets under the top sheet of paper. The sensitivity test also determined the EDPL results increase in quality as the amount of dust residue decreases on the surface.

Forensic Medicine↗

Contact dermatitis from telefax paper.

A nonatopic female secretary developed hand dermatitis after 1 year of full-time work with telefax paper. Her dermatitis improved on sick leave and vacation and relapsed at work. Patch testing showed allergic reactions to several fragrances, balsam of Peru, colophony and neomycin. In 3 patch test sessions, her own telefax papers gave a doubtful reaction which persisted for 4 days, but it could not be determined whether the reactions were allergic or irritant. The colophony content of the telefax paper was about 1%, and it was possible that colophony in the telefax paper was responsible for the weak path test reactions. Accordingly, the contact dermatitis would have been allergic. The patient has now been relocated, does not handle telefax paper, and is symptomless. We concluded that our patient had an occupational hand contact dermatitis induced by telefax paper and possibly caused by colophony allergy.

Dermatitis, Allergic Contact↗