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Virgibacillus carmonensis sp. nov., Virgibacillus necropolis sp. nov. and Virgibacillus picturae sp. nov., three novel species isolated from deteriorated mural paintings, transfer of the species of the genus salibacillus to Virgibacillus, as Virgibacillus marismortui comb. nov. and Virgibacillus salexigens comb. nov., and emended description of the genus Virgibacillus.

A group of 13 strains was isolated from samples of biofilm formation on the mural paintings of the Servilia tomb (necropolis of Carmona, Spain) and the Saint-Catherine chapel (castle at Herberstein, Austria). The strains were subjected to a polyphasic taxonomic study, including (GTG)5-PCR, 16S rDNA sequence analysis, DNA-DNA hybridizations, DNA base ratio determination, analysis of fatty acids, polar lipids and menaquinones and morphological and biochemical characterization. In a phylogenetic tree based on neighbour-joining of 16S rDNA sequences, the strains are divided in two major groups, representing three novel species according to DNA-DNA relatedness, that are positioned at approximately equal distances from Virgibacillus and Salibacillus. After comparison of the novel results with existing data, the transfer of the species of Salibacillus to Virgibacillus is proposed, with the resulting new combinations Virgibacillus marismortui comb. nov. and Virgibacillus salexigens comb. nov. Additionally, three novel species are described, for which the names Virgibacillus carmonensis sp. nov., Virgibacillus necropolis sp. nov. and Virgibacillus picturae sp. nov. are proposed. The respective type strains are LMG 20964T (=DSM 14868T), LMG 19488T (=DSM 14866T) and LMG 19492T (= DSM 14867T). Finally, an emended description of the genus Virgibacillus is given.

Bacillus↗

Paenibacillus sepulcri sp. nov., isolated from biodeteriorated mural paintings in the Servilia tomb.

In 2001, a Gram-variable, facultatively anaerobic, endospore-forming bacterium isolated from biodeteriorated mural paintings in the Servilia tomb of the Roman necropolis of Carmona was deposited as Paenibacillus strain LMG 19508. Subsequently, the strain was characterized in detail using phenotypic and molecular methods. The 16S rRNA gene sequence confirmed that the strain belongs to the genus Paenibacillus and indicated its relationship to Paenibacillus mendelii CCM 4839(T) (96.7 % sequence similarity). The predominant menaquinone was MK-7. The cell wall contained meso-diaminopimelic acid of the A1gamma type. The DNA G+C content (50 mol%) and the major fatty acid (anteiso-C(15 : 0)) of strain LMG 19508(T) were also consistent with its affiliation to the genus Paenibacillus. DNA-DNA hybridization distinguished strain LMG 19508(T) from other phylogenetically related Paenibacillus species. Therefore, the isolate represents a novel species, for which the name Paenibacillus sepulcri sp. nov. is proposed. The type strain is CCM 7311(T) (=LMG 19508(T)).

Bacteria, Anaerobic↗

The use of fatty acid methyl ester analysis (FAME) for the identification of heterotrophic bacteria present on three mural paintings showing severe damage by microorganisms.

Mural paintings in Carmona (Spain), Herberstein (Austria) and Greene (Germany), showing visible deterioration by microorganisms, were sampled to investigate the biodiversity of the heterotrophic bacteria present. Four hundred twenty-eight bacterial strains were isolated from which 385 were characterized by fatty acid methyl ester analysis (FAME). The isolates were grouped into 41 clusters on the basis of their FAME profiles, 20 isolates remained ungrouped. The majority (94%) of the isolates comprised the gram-positive bacteria and the main clusters were identified as Bacillus sp., Paenibacillus sp., Micrococcus sp., Arthrobacter sp. and Staphylococcus sp. Other clusters contain nocardioform actinomycetes and gram-negative bacteria, respectively. A cluster of the latter contained extreme halotolerant bacteria isolated in Herberstein. The FAME profiles of this cluster showed a high similarity with Halomonas.

Austria↗

Occupational dermatoses in a plant producing binders for paints and glues.

A survey of occupational dermatoses, based on a questionnaire, clinical examination and patch testing, was carried out among present and former employees in a plant producing binders for glues and paints. The binders are chemically based on vinyl acetate and/or acrylates. Preservatives are needed and those based on methylchloroisothiazolinone/methylisothiazolinone (MCI/MI) are the most commonly used. The questionnaire was answered by 83 out of 85 present employees (97.6%) and 12 out of 15 former employees (80.0%). 76 of the present (89.4%) and 11 of the former employees (73.3%), respectively, participated in the clinical examination and patch testing, which was conducted with a standard test series and chemicals from the work environment. Occupational dermatosis was diagnosed in 22 present workers (28.9%). Irritant and allergic contact dermatitis was demonstrated in 9 (11.8%) and 13 (17.1%) employees, respectively. Occupational contact allergy to acrylates and formaldehyde was detected in 3 workers and 1 worker, respectively. 12 individuals (9 present and 3 former production workers) had an occupational contact allergy to MCI/MI. 4 of the present workers had spilled Kathon LX on their skin resulting in chemical burns and allergic contact dermatitis. In total, the figure for occupational skin diseases among all present production workers was 40.4%.

Acrylates↗

Volatile organic compound emissions from latex paint--Part 2. Test house studies and indoor air quality (IAQ) modeling.

Emission models developed using small chamber data were combined with an Indoor Air Quality (IAQ) model to analyze the impact of volatile organic compound (VOC) emissions from latex paint on indoor environments. Test house experiments were conducted to verify the IAQ model's predictions. The agreement between model predictions and experimental measurements met the American Society for Testing and Materials criteria for model verification in the room with the source and met most of the requirements in other rooms. The major cause of disagreement between the model predictions and the experimental data in the test house appears to be an inadequate sink model.

Air Pollution, Indoor↗

Visualization of jet flows over a plate by pressure-sensitive paint experiments and comparison with CFD.

Flow fields created by underexpanded sonic jets impinging on an inclined flat plate were studied experimentally using the pressure sensitive paint (PSP) measurement technique. The measurement system and some representative results are presented here. Two binders, thin-layer chromatography (TLC) plates and anodized aluminum (A-A) plates were tested with bathophen ruthenium chloride as a luminophore. The results show that both the binders can be used. TLC plates are preferable because their luminescent intensity is almost twice that of the A-A plates. Quantitative measurements require accurate temperature calibration. A preliminary effort to elucidate the flow structure by combining the PSP results with a computer simulation of the same flow field is presented. Although good agreement is obtained between the experimental and numerical results, future quantitative comparisons are necessary to yield a useful tool in the analysis of the jet-plate interaction flows.

Aluminum↗

Automobile paint effective as an insect attractant.

Two acrylic auto paints were effective attractants for sap beetles, Stelidota geminata (Say) and Glischrochilus fasciatus (Oliver). Response to the synthetic lures was sufficient to encourage additional testing of their potential in survey and control programs.

Acrylic Resins↗

Detection of indigenous Halobacillus populations in damaged ancient wall paintings and building materials: molecular monitoring and cultivation.

Several moderately halophilic gram-positive, spore-forming bacteria have been isolated by conventional enrichment cultures from damaged medieval wall paintings and building materials. Enrichment and isolation were monitored by denaturing gradient gel electrophoresis and fluorescent in situ hybridization. 16S ribosomal DNA analysis showed that the bacteria are most closely related to Halobacillus litoralis. DNA-DNA reassociation experiments identified the isolates as a population of hitherto unknown Halobacillus species.

Austria↗

Normal serum activities of liver enzymes in Swedish paint industry workers with heavy exposure to organic solvents.

The serum activities of the liver enzymes alanine aminotransferase, aspartate aminotransferase, ornithine carbamyl transferase, and gamma-glutamyl transferase were examined in 47 paint industry workers and unexposed age matched referents. The workers were exposed to a mixture of industrial solvents, of which xylene was the main component in most cases. The median total exposure was about 50% of Swedish 1981 threshold limit values according to measurements of individual solvent exposure performed at the same time. No differences in enzyme activities were shown either when the whole exposed and referent groups were compared or when the five workers with outstanding solvent exposures of five times the TLV or more were compared with their referents. It is concluded that in most workers the liver seems to remain largely undamaged from inhalation exposure to a commonly used mixture of non-chlorinated solvents. In many workers this seems to hold true even for high exposures for limited periods.

Adult↗

Retrospective assessment of solvent exposure in paint manufacturing.

This paper describes how exposure to solvents at two large paint making sites was assessed in a study carried out to investigate the possibility of neuropsychological effects resulting from long term exposure to organic solvents. A job exposure matrix was constructed by buildings and year. A detailed plant history was taken and this was used to identify uniform exposure periods during which workers' exposure to solvents was not thought to have changed significantly. Exposure monitoring data, collected by the company before the study, was then used to characterise exposure within each uniform exposure period. Estimates were made for periods during which no air monitoring was available. Individual detailed job histories were collected for subjects and controls. The job histories were used to estimate exposure on an individual basis with the job exposure matrix. Exposure was expressed as duration, cumulative dose, and intensity of exposure. Classification of exposure by duration alone was found to result in misclassification of subjects.

Environmental Monitoring↗

Pulmonary toxicity of components of textile paint linked to the Ardystil syndrome: intratracheal administration in hamsters.

OBJECTIVES: It was hypothesised from an epidemiological investigation that a formula change from Acramin FWR (a polyurea) to Acramin FWN (a polyamide-amine) had led to severe pulmonary disease in textile printing sprayers in SPAIN AND ALGERIA. To verify this, the pulmonary toxicity of the components of the paint systems involved was assessed in experimental animals. METHODS: Individual components and relevant mixtures, diluted in phosphate buttered saline, were given by intratracheal instillation of 2 ml/kg to hamsters. Pulmonary toxicity was assessed on days 3, 7, 14, 28, and 92 after a single intratracheal instillation, by histology and by measuring wet and dry lung weight, protein concentration, the activities of lactate dehydrogenase, alkaline phosphatase, beta-N-acetyl-glucosaminidase, and gamma-glutamyltransferase, inflammatory cell number and distribution in bronchoalveolar lavage fluid (BALF), and hydroxyproline content in dried lung tissue. RESULTS: Based on the doses that killed 50% of the animals (LD50s), the various components were found to be 10 to 1250 times more toxic when given intratracheally than when given orally (according to reported oral LD50s in rats). Acramin FWN, Acramin FWR, Acrafix FHN, or their mixtures caused lung damage. Protein concentration, enzyme activities, total cell number, and percentage of polymorphonuclear neutrophils were increased in BALF during the first week after intratracheal instillation. Lung weights remained high for at least a month. Histology showed inflammatory cell infiltration and subsequent fibrosis with collagen deposition. This finding was confirmed by an increased hydroxyproline content in dried lung tissue. Acramoll W did not show toxic effects. CONCLUSIONS: The study suggests that there is no major difference, in hamsters, between the acute intratracheal toxicity of Acramin FWR and that of Acramin FWN. Consequently, there is no simple toxicological explanation for the epidemiological hypothesis. However, the pulmonary toxicity of these non-irritant polymeric compounds is surprisingly high. The Ardystil disaster and these results should serve as a strong warning that conventional toxicity testing of chemicals does not necessarily protect workers against respiratory toxicity.

Acetylglucosaminidase↗

A complete set of repeat-depleted, PCR-amplifiable, human chromosome-specific painting probes.

We describe the generation of a complete set of human chromosome-specific painting probes depleted in repetitive sequences. These probes yield highly specific signals when hybridized without the addition of a blocking agent, such as Cot-1 DNA, and without probe preannealing prior to hybridization. Fluorescent intensities and signal-to-background ratios for these probes are comparable to those of untreated probes hybridized with Cot-1 DNA. We demonstrate the suitability of these probes for applications with very complex probe sets, such as multiplex-FISH.

Binding, Competitive↗

Micro- and macrochromosome paints generated by flow cytometry and microdissection: tools for mapping the chicken genome.

Despite the chicken being one of the most genetically mapped of all animals, its karyotype remains poorly defined. This is primarily due to microchromosomes that belie assignment by conventional methods. To address this problem, we have developed chromosome-specific paints using flow cytometry and microdissection. For the microchromosomes it was necessary to amplify and label DNA from single microdissected chromosomes.

Animals↗

Generation of whole-chromosome painting probes specific to each chicken macrochromosome.

The realization of physical and genetic maps of the chicken genome is dependent on progress in cytogenetic knowledge of its karyotype. To help achieve this goal, we constructed amplified representative DNA samples of the chicken chromosomes 1, 2, 3, 4, 5, 6, 7, 8, Z, W and of the terminal heterochromatin part of Zq by chromosome microdissection and DOP-PCR amplification. These chromosome DNA samples, which represent about 75% of the chicken genome, were used to generate whole chromosome painting probes for FISH. The direct application of these chromosome specific probes is dual FISH localization and characterization of panels of chicken interspecific somatic hybrids. We discuss some aspects of the chicken genome and its repeated sequences.

Animals↗

Molecular cytotaxonomy of New World monkeys (Platyrrhini) - comparative analysis of five species by multi-color chromosome painting gives evidence for a classification of Callimico goeldii within the family of Callitrichidae.

Chromosome rearrangements are considered as "rare genomic changes" and can provide useful markers and even landmarks for reconstructing phylogenies complementary to DNA sequence data and bio-morphological comparisons. Here, we applied multi-directional chromosome painting to reconstruct the chromosome phylogeny and evolutionary relationships among the New World monkey (Platyrrhini) species Callithrix argentata, Cebuella pygmaea, Saguinus oedipus, Callithrix jacchus and Callimico goeldii. The results clarified several aspects of New Wold monkey phylogeny. In particular the phylogenetic position of C. goeldii was elucidated, which has been controversially discussed and variously classified in the family Callitrichidae, in the family Cebidae or in its own family Callimiconidae. Comparative genome maps were established by multi-color fluorescence in situ hybridization (FISH) with human, S. oedipus and Lagothrix lagothricha chromosome- specific DNA probes. From these data we reconstructed the putative ancestral karyotype of all Callitrichidae. Various derived chromosomal syntenies are shared by all five species and cytogenetically define Callitrichidae - including Callimico goeldii -- as a distinctive group within the Platyrrhini. C. pygmaea and C. argentata share identical chromosomal syntenies from which S. oedipus and C. jacchus differ by single independent translocations. A common derived chromosomal change links Callimico with the marmosets to the exclusion of the tamarins, however, it has further diverged from an ancestral marmoset karyotype by at least four apomorphic rearrangements. Saimiri sciureus, representing the Cebinae, exclusively shares a derived syntenic association with all Callithrichidae, defining the genus Saimiri as a sister group.

Animals↗

A new balanced autosomal reciprocal translocation in cattle revealed by banding techniques and human-painting probes.

Three hundred and twenty-two (264 males and 58 females), randomly sampled Grey Alpine cattle individuals from Northeastern Italy, were investigated cytogenetically by both conventional chromosome staining and R-banding. Two hundred and eighty-one (87%) individuals had a normal karyotype and 41 (13%) carried chromosomal aberrations such as (a) rob(1;29) in two individuals, (b) rob(26;29) in 36 individuals, (c) XX/XY-chimerism in two individuals, and (d) an abnormally long chromosome in one individual. All these aberrations except (d) have been described before. GBG-, RBG-, CBA-banding and sequential GBG/CBA- and RBG/CBA-banding techniques revealed that the abnormally long chromosome was the result of a reciprocal translocation between chromosomes 1 (q21-->qter) and 5 (q11-->q33), as confirmed also by chromosome painting with human chromosome 3 and 12 probes. The dam of the carrier bull carried the same translocation, while the grandam showed a normal karyotype. Since the sire of the dam was not available for study, no conclusion about the origin of the chromosome translocation could be drawn. The carrier bull was eliminated because of poor fertility. The dam had three other calves, which all were chromosomally normal. On average the dam had to be served 2.5 times (breed average was 1.2) to be in calf.

Animals↗

Cross-species chromosome painting in the Perissodactyla: delimitation of homologous regions in Burchell's zebra (Equus Burchellii) and the white (Ceratotherium Simum) and black rhinoceros (Diceros Bicornis).

Conserved chromosomal segments in the black rhinoceros, Diceros Bicornis (DBI, 2n = 84), and its African sister-species the white rhinoceros, Ceratotherium Simum (CSI, 2n = 82), were detected using Burchell's zebra (Equus Burchellii, EBU, 2n = 44) chromosome-specific painting probes supplemented by a subset of those developed for the horse (Equus Caballus, ECA, 2n = 64). In total 41 and 42 conserved autosomal segments were identified in C. Simum and D. Bicornis respectively. Only 21 rearrangements (20 fissions and 1 fusion) are necessary to convert the Burchell's zebra karyotype into that of the white rhinoceros. One fission distinguishes the D. Bicornis and C. Simum karyotypes which, excluding heterochromatic differences, are identical in all respects at this level of resolution. Most Burchell's zebra chromosomes correspond to two rhinoceros chromosomes although in four instances (EBU18, 19, 20 and 21) whole chromosome synteny has been retained among these species. In contrast, one rhinoceros chromosome (DBI1, CSI1) comprises two separate Burchell's zebra chromosomes (EBU11 and EBU17). In spite of the high diploid numbers of the two rhinoceros species their karyotypes are surprisingly conserved offering a glimpse of the putative ancestral perissodactyl condition and a broader understanding of genome organization in mammals.

Animals↗