Evaluation of interference to conventional and real-time PCR for detection and quantification of fungi in dust

Advances in polymerase chain reaction (PCR) have permitted accurate, rapid and quantitative identification of microorganisms in pure cultures regardless of viability or culturability. In this study, a simple sample processing method was investigated for rapid identification and quantification of fungal spores from dust samples using both conventional and real-time PCR. The proposed method was evaluated for susceptibility to interference from environmental dust samples. Stachybotrys chartarum and Aspergillus fumigatus were used as test organisms. The sensitivity of detection in pure culture was 0.1 spore DNA equivalents per PCR reaction corresponding to 20 spores ml(-1) in the sample. However, 1 spore DNA equivalent per PCR reaction corresponding to 200 spores ml(-1) in the sample was the lowest amount of spores tested without interference in dust samples spiked with spores of either fungal species. The extent of inhibition was calculated using conventional and real-time PCR reactions containing fungal spores, specific primers, specific probes (for real-time PCR) and various amounts of dust. The results indicate that the extent of inhibition by dust on PCR varies with the type and amount of dust, and number of spores. No interference in the analysis of spiked samples was detected from 0.2 mg ml(-1) of four real-life dust samples at p-value >0.05 using 2 x 10(4) spores for conventional PCR and 2 x 10(5) spores for real-time PCR. However, samples containing >0.2 mg ml(-1) real-life dust compromised the PCR assay. These results suggest the potential usefulness of a simple sample processing method in conjunction with PCR for monitoring the fungal content of aerosols collected from indoor environments.     

Pool of Dust Particles over the Asian Continent: Balloon-borne Optical Particle Counter and Ground-based Lidar Measurements at Dunhuang, China

Measurements of aerosols were made in 2001 and 2002 at Dunhuang (40 degrees 00'N, 94 degrees 30'E), China to understand the nature of atmospheric particles over the desert areas in the Asian continent. Balloon-borne measurements with an optical particle counter suggested that particle size and concentration had noticeable peaks in super micron size range not only in the boundary mixing layer but also in the free troposphere. Super-micron particle concentration largely decreased in the mid tropopause (from 5 to 10 km; above sea level, a.s.l.). Lidar measurements made during August 2002 at Dunhuang suggested the possibility that mixing of dust particles occurred from near the ground to about 6 km even under calm weather conditions, and a large depolarization ratio of particulate matter was found in the aerosol layer. The top of the aerosol layer was found at heights of nearly 6 km (a.s.l.). It is strongly suggested that nonspherical dust particles (Kosa particles) frequently diffused in the free atmosphere over the Taklamakan desert through small-scale turbulences and are possible sources of dust particles of weak Kosa events that have been identified in the free troposphere not only in spring but also in summer over Japanese archipelago. Electron microscopic experiments of the particles collected in the free troposphere confirmed that coarse and nonspherical particles observed by the mineral particle were major components of coarse mode (diameter larger than 1 microm) below about 5 km over Dunhuang, China.     

Total versus inhalable sampler comparison study for the determination of asphalt fume exposures within the road paving industry

Exposure to asphalt fumes has a threshold limit value (TLV of 0.5 mg m(-3) (benzene extractable inhalable particulate) as recommended by the American Conference of Governmental Industrial Hygienists (ACGIH). This reflects a recent change (2000) whereby two variables are different from the previous recommendation. First is a 10-fold reduction in quantity from 5 mg m(-3) to 0.5 mg m(-3). Secondly, the new TLV specifies the "inhalable" fraction as compared to what is presumed to be total particulate. To assess the impact of these changes, this study compares the differences between measurements of paving asphalt fume exposure in the field using an "inhalable" instrument versus the historically used 'total' sampler. Particle size is also examined to assist in the understanding of the aerodynamic collection differences as related to asphalt fumes and confounders. Results show that when exposures are limited to asphalt fumes, a 1:1 relationship exists between samplers, showing no statistically significant differences in benzene soluble matter (BSM). This means that for the asphalt fume ACGIH TLV, the 'total' 37-mm sampler is an equivalent method to the "inhalable" method, referred to as IOM (Institute of Occupational Medicine), and should be acceptable for use against the TLV. However, the study found that when confounders (dust or old asphalt millings) are present in the workplace, there can be significant differences between the two samplers' reported exposure. The ratio of IOM/Total was 1.37 for milling asphalt sites, 1.41 for asphalt paving over granular base, and 1.02 for asphalt over asphalt pavements.     

Utility of dynamics as indicators of stress in population models

The relationships between the dynamics of environmentally and chemically stressed populations and indicators of the effects of the stressor are explored in a model framework. The physiologically structured population, represented by a system of McKendrick–von Foerster hyperbolic partial differential equations, includes the dynamics of numerous individuals distinguished by ecotype. Chemical uptake of nonpolar narcotics is modeled by first order kinetics. Classical methodologies, frequency analysis and phase space reconstruction, are explored in a search for indicators of magnitude of stress. When these techniques proved generally unsuccessful for the objective of indicator selection in our model setting, summary statistics, as related to bifurcation diagrams, were constructed and appear more useful as indicators. It is concluded that physiological structures generally lead to more feasible measurable indicators of magnitude of stress than do specifics of population dynamics.     

Fuel characteristics and emissions from biomass burning and land-use change in Nigeria

Nigeria is one of the 13 low-latitude countries that have significant biomass burning activities. Biomass burning occurs in moist savanna, dry forests, and forest plantations. Fires in the forest zone are associated with slash-and-burn agriculture; the areal extent of burning is estimated to be 80% of the natural savanna. In forest plantations, close to 100% of litter is burned. Current estimates of emissions from land-use change are based on a 1976 national study and extrapolations from it. The following non-carbon dioxide (CO₂) trace gas emissions were calculated from savanna burning: methane (CH₄), 145 gigagrams (Gg); carbon monoxide (CO), 3831 Gg; nitrous oxide (N₂O), 2 Gg; and nitrogen oxides (NO_x), 49 Gg. Deforestation rates in forests and woodlands are 300 × 10³ ha (kilohectare, or kha) and 200 × kha per year, respectively. Trace gas emissions from deforestation were estimated to be 300 Gg CH₄, 2.4 Gg N₂O, and 24 Gg NO_x. CO₂ emissions from burning, decay of biomass, and long-term emissions from soil totaled 125 561 Gg. These estimates should be viewed as preliminary, because greenhouse gas emission inventories from burning, deforestation, and land-use change require two components: fuel load and emission factors. Fuel load is dependent on the areal extent of various land uses, and the biomass stocking and some of these data in Nigeria are highly uncertain.     

Life-history parameters and vertical distribution of Maurolicus muelleri in Masfjorden in summer

Population characteristics, individual life-history variables, feeding and vertical distribution of the mesopelagic fish Müller's pearlside Maurolicus muelleri collected in 1990 in Masfjorden, western Norway, are reported as well as environmental variables from the fjord. Minimum size at maturity was far smaller than reported from previous investigations in the same region. Fecundity was size-dependent and total egg numbers were higher than reported from other investigations of M. muelleri world wide, while the number of maturing eggs was far lower than observed in the same region earlier. Food concentration in the fjord was an order of magnitude lower than previous early summer observations, and several factors indicated that feeding opportunities may have been low for a long period. Daily feeding rate (g prey g^-1 fish) decreased with increasing fish size. These observations fit well with a model of maximizing fitness by means of a flexible size at maturity. Minimum age at maturity seems to be achieved at the expense of fecundity. M. muelleri was concentrated in a 20 to 30 m deep sound scattering layer (SSL). The SSL stayed close to the surface during the night and at 100 to 180 m during the daytime. The vertical position of the SSL varied instantaneously with changes in surface light intensity, remaining at 10^-3 to 10^-4 mol m^-1 s^-1 at the top of the SSL. Stomach fullness was highest during the night; feeding intensity seems to have been peaked at dusk. Cladocerans were the main prey ranked by number, copepods by biomass. Intake of large copepods increased with fish size.     

Genetic population structure of the recently introduced Asian clam,Potamocorbula amurensis,in San Francisco Bay

The genetic population structure of the recently introduced Asian clam, Potamocorbula amurensis, in San Francisco Bay was described using starch gel electrophoresis at eight presumptive loci. Specimens were taken from five environmentally distinct sites located throughout the bay. The population maintains a high degree of genetic variation, with a mean heterozygosity of 0.295, a mean polymorphism of 0.75, and an average of 3.70 alleles per locus. The population is genetically homogeneous, as evidenced from genetic distance values and F-statistics. However, heterogeneity of populations was indicated from a contingency chi-square test. Significant deviations from Hardy-Weinberg equilibrium and heterozygote deficiencies were found at the Lap-1 locus for all populations and at the Lap-2 locus for a single population. High levels of variability could represent a universal characteristic of invading species, the levels of variability in the source population(s), and/or the dynamics of the introduction. Lack of differentiation between subpopulations may be due to the immaturity of the San Francisco Bay population, the general purpose phenotype genetic strategy of the species, high rates of gene flow in the population, and/or the selective neutrality of the loci investigated.     

Comparison of allozyme and mitochondrial DNA spatial differentiation in the limpet Patella vulgata

Allozyme and restriction enzyme analysis of mtDNA was used to study variation in samples from British populations of the marine limpet Patella vulgata in two regions. South Wales and Northeast England. Allozyme analysis revealed significant differences in allele frequencies among samples. However F _ST (population differentiation) values were no higher between than within regions, indicating that genetic heterogeneity was localised and not related to geographic separation. For mtDNA, samples from South Wales exhibited higher haplotype diversity values than samples from Northeast England. In addition there were substantial differences in the haplotype distribution between regions. The value of , the haploid analogoue of F _ST , was low within regions (=0.09) but high between regions (=0.44). The estimated difference in migration rate for allozymes and mtDNA exceed the neutral expectation, unless it is assumed that there are influential differences in the magnitude of female and male gametic dispersal.     

Ecoprofit-Styria-prepare Thirteen pollution prevention case studies in the Styrian industry

The paper focuses upon the organization of a federal state-funded pollution prevention project in the Styrian industry. The project includes 13 companies from the textile, pulp and paper, machine building, wood working and printed circuit board manufacturing industries, covering most of the sectors and sizes in the Styrian industry. It started in January 1994 and will last for one year. It will demonstrate the possibilities of pollution prevention and the need for further research work. This project will make use of the methods and tools that were refined in the Austrian Prepare project. As a first step, a systematic balance of all the inputs and outputs of a company is made, after which the weak points and inefficiencies of material and energy use are identified and the options for improvements, both economical and ecological, are defined. Consequently, modifications in products and production lead to a situation with less waste and emissions. The preliminary lessons from these projects are presented: as a rule, the utilities (consumption of process materials and water, cleaning, energy, preparatory and finishing steps) are treated as black boxes and usually represent a considerable optimization potential. In these areas especially there is usually a lack of information and coordination as well as a need for a systematic and comprehensive approach. Leadership in the company and creative consultants are needed for starting lasting successful pollution prevention projects.