Profile of the culturable microbiome capable of producing acyl-homoserine lactone in the tobacco phyllosphere

Bacterial populations coexisting in the phyllosphere niche have important effects on plant health. Quorum sensing (QS) allows bacteria to communicate via diffusible signal molecules, but QS-dependent behaviors in phyllosphere bacterial populations are poorly understood. We investigate the dense and diverse N-acyl-homoserine lactone (AHL)-producing phyllosphere bacteria living on tobacco leaf surfaces via a culture-dependent method and 16S rRNA gene sequencing. Our results indicated that approximately 7.9%-11.7% of the culturable leaf-associated bacteria have the ability to produce AHL based on the assays using whole-cell biosensors. Sequencing of the 16S rRNA gene assigned the AHL-producing strains to two phylogenetic groups, with Gammaproteobacteria (93%) as the predominant group, followed by Alphaproteobacteria. All of the AHL-producing Alphaproteobacteria were affiliated with the genus Rhizobium, whereas the AHL-producing bacteria belonging to the Gammaproteobacteria mainly fell within the genera Pseudomonas, Acinetobacter, Citrobacter, Enterobacter, Pantoea and Serratia. The bioassays of supernatant extracts revealed that a portion of the strains have a remarkable AHL profile for AHL induction activity using the two different biosensors, and one compound in the active extract of a representative isolate, NTL223, corresponded to 3-oxo-hexanoyl-homoserine lactone. A large population size and diversity of bacteria capable of AHL-driven QS were found to cohabit on leaves, implying that cross-communication based AHL-type QS may be common in the phyllosphere. Furthermore, this study provides a general snapshot of a potential valuable application of AHL-producing bacteria inhabiting leaves for their presumable ecological roles in the phyllosphere.     

Study on nitrile-degrading microorganisms

Two strains of bacteria were isolated from nitrile polluted soils, and identified as Corynebacterium boffmanii and Arthrobacter flavescens. Acetonitrile, propionitrile, butyronitrile and acrylonitrile were degraded by these bacteria to yield corresponding amides, carboxylic acids and ammonia. The nitrile-degrading abilities of these strains were investigated. The removal rates for the nitrile were nearly 100%, after these bacteria were grown in medium containing 10000 ppm of aceto-, propio-, or butyronitrile at 28 ℃ for 24h. When the reaction mkture consisting of 5000 ppm of above mentioned nitriles or acrylonitrile and 20g (dry cell) /L resting cells of the two strains in 0.06mol/L phosphate buffer (pH7.5) was incubated separately at 25 ℃ with moderate shaking for 15 or 45 min, the nitrile could be degraded completely. The optimum growth conditions for C.hoffmanii and A.flavescens were studied as well.     

Biogeochemical cyclic activity of bacterial arsB in arsenic-contaminated mines

Biogeochemical cyclic activity of the ars (arsenic resistance system) operon is arsB influx/effux encoded by the ecological of Pseudomonas putida.This suggests that studying arsenite-oxidizing bacteria may lead to a better understanding of molecular geomicrobiology,which can be applied to the bioremediation of arsenic-contaminated mines.This is the first report in which multiple arsB-binding mechanisms have been used on indigenous bacteria.In ArsB (strains OS-5; ABB83931; OS-19; ABB04282 and RW-28; ABB88574...     

Adaptability of microbial inoculators and their contribution to degradation of mineral oil and PAHs

Five dominant bacteria strains（A cetobacter sp., Alcdigenes sp., Micrococcus sp., Arthrobacter sp. and Bacillus sp.） and five fungi strains （Cephalosporium sp. Ⅰ, Cephalosporlum sp. Ⅱ, Aspergillus sp. Ⅰ, Aspergillus sp. Ⅱ and Fusarium sp.） isolated from petroleum-contaminated soil were used to assess the potential capability of mineral oil and PAH enhanced degradation separately and jointly using the batch liquid medium cultivation with diesel oil spiked at 1000 mg/L. The experiment was performed on a reciprocal shaker in the darkness at 25℃ to 30℃ for 100 d. The dynamic variation in the activity of microbial inoculators in each treatment and the degradation of the target pollutants during the period of experiment were monitored. Results showed a more rapid biodegradation of mineral oil and PAHs at the beginning of the experiment （about 20 d） by dominant bacteria, fungi and their mixture than that of the indigenous microorganisms, however, thereafter an opposite trcnd was exhibited that the removal ratio by indigenous microorganisms was superior to any other dominant treatments and the tendency lasted till the end of the experiment, indicating the limited competitive capability of dominant microorganisms to degrade the contaminants, and the natural selection of indigenous microorganisms for use in the removal of the contaminants. At the end of the experiment, the removal ratio of mineral oil ranged from 56.8 % to 79.2 % and PAHs ranged from 96.8 % to 99.1% in each treatment by microbial inoculators,     

Effects of imazethapyr spraying on plant growth and leaf surface microbial communities in Arabidopsis thaliana

Imazethapyr (IM) is an acetolactate synthase (ALS)-inhibiting herbicide that has been widely used in recent years. However, IM spraying can lead to the accumulation of herbicide residues in leaves. Here, we determined the effects of IM spraying on the plant growth and leaf surface microbial communities of Arabidopsis thaliana after 7 and 14?days of exposure. The results suggested that IM spraying inhibited plant growth. Fresh weight decreased to 48% and 26% of the control value after 7 and 14?days, respectively, of 0.035?kg/ha IM exposure. In addition, anthocyanin content increased 9.2-fold and 37.2-fold relative to the control content after 7 and 14?days of treatment, respectively. Furthermore, IM spraying destroyed the cell structures of the leaves, as evidenced by increases in the number of starch granules and the stomatal closure rate. Reductions in photosynthetic efficiency and antioxidant enzyme activity were observed after IM spraying, especially after 14?days of exposure. The diversity and evenness of the leaf microbiota were not affected by IM treatment, but the composition of community structure at the genus level was altered by IM spraying. Imazethapyr application increased the abundance of Pseudomonas, a genus that includes species pathogenic to plants and humans, indicating that IM potentially increased the abundance of pathogenic bacteria on leaves. Our findings increase our understanding of the relationships between herbicide application and the microbial community structures on plant leaves, and they provide a new perspective for studying the ecological safety of herbicide usage.     

Spatial analysis of plant detritus processing in a Mediterranean river type: the case of the River Tirso Basin,Sardinia, Italy

The river continuum concept represents the most general framework addressing the spatial variation of both structure and function in river ecosystems.In the Mediterranean ecoregion,summer drought events and dams constitute the main sources of local disturbance to the structure and functioning of river ecosystems occurring in the river basin.In this study,we analysed patterns of spatial variation of detritus processing in a 7th order river of the Mediterranean ecoregion(River Tirso,Sardinia0Italy)and in three 4th order sub-basins which were exposed to different summer drought pressures.The study was carried out on phragmites australis and Alnus glutionsa leaf detritus at 31 field sites in seasonal field experiment Detritus processing rates were higher for Alnus glutionsa than for Phragmites australis plant detritus.Processing rates of Alnus glutionsa leaves varied among seasons and study sites from 0.006d^-1 to 0.189d^-1 and those of Phragmites australis leaves ranged from 0.0008d^-1 to 0.102d^-1,with the lowest values occurring at sites exposed to summer drought.Seasons and sites accounted for a significant proportion of such variability.Alder detritus decay rates generally decreased with increasing stream order,while reed detritus decay rates generallyincreased on the same spatial gradient.Summer drought events affected these spatial patterns of variation by influencing significantly the decay rates of both plant detritus.The comparisons among and within sub-basins showed strong negative influence of summer drought on detritus processing rates.Similarly,in the entire River Tirso basin decay rates were always lower at disturbed than at undisturbed sites for each stream order;decay rates of reed detritus remained lower at those sites even after the end of the disturbance events,while alder decay rates recovered rapidly from the summer drought perturbations.The different recovery of the processing rates of the two leaves could also explain the different patterns of spatial variation observed between the two leaves.     

Isolation and characterization of gasoline-degrading bacteria from gas station leaking-contaminated soils

The effects of culture conditions in vitro and biosurfactant detection were studied on bacterial strains capable of degrading gasoline from contaminated soils near gas station. The main results were summarized as follows. Three bacteria （strains Q10, Q14 and Q18） that were considered as efficiently degrading strains were isolated and identified as Pseudomonas sp., Flavobaeterium sp. and Rhodococcus sp., respectively. The optimal growth conditions of three bacteria including pH, temperature and the concentration of gasoline were similar. The reduction in surface tension was observed with all the three bacteria, indicating the production of biosurfactant compounds. The value of surface tension reduced by the three strains Q10, Q14 and Q18 was 32.6 mN.m, 12.4 mN. m and 21.9 mN.m, respectively. Strain Q10 could be considered as a potential biosurfactant producer. Gasoline, diesel oil, benzene, toluene, ethylbenzene and xylene （BTEX） could easily be degraded by the three isolates. The consortium was more effective than the individual cultures in degrading added gasoline, diesel oil, and BTEX. These results indicate that these strains have great potential for in situ remediation of soils contaminated by gas station leaking.     

Screening on oil-decomposing microorganisms and application in organic waste treatment machine

As an oil-decomposable mixture of two bacteria strains ( Bacillus sp. and Pseudomonas sp. ), Y3 was isolated after 50 d domestication under the condition that oil was used as the limited carbon source. The decomposing rate by Y3 was higher than that by each separate individual strain, indicating a synergistic effect of the two bacteria. Under the conditions that T = 25-40℃, pH = 6--8, HRT (Hydraulic retention time) = 36 h and the oil concentration at 0.1%,Y3 yielded the highest decomposing rate of 95.7 %. Y3 was also applied in an organic waste treatment machine and a certain rate of activated bacteria was put into the stuffing. A series of tests including humidity, pH, temperature, C/N rate and oil percentage of the stuffing were carried out to check the efficacy of oil-decomposition. Results showed that the oil content of the stuffing with inoculums was only half of that of the control. Furthermore, the bacteria were also beneficial to maintain the stability of the machine operating. Therefore, the bacteria mixture as well as the machines in this study could be very useful for waste treatment.     

Physicochemical characteristics of ambient particles settling upon leaf surfaces of urban plants in Beijing

Particulate pollution is a serious health problem throughout the world, exacerbating a wide range of respiratory and vascular illnesses in urban areas. Urban plants play an important role in reducing particulate pollution. Physicochemical characteristics of ambient particles settling upon leaf surfaces of eleven roadside plants at four sites of Beijing were studies. Results showed that density of particles on the leaf surfaces greatly varied with plant species and traffic condition. Fraxinus chiuensis, Sophora japonica Ailanthus altissima, Syringa oblata and Prunus persica, had larger densities of particles among the tall species. Due to resuspension of road dust, the densities of particles of Euonymus japonicns and Parthenocissus quinquefolia with low sampling height were 2-35 times to other taller tree species. For test plant species, micro-roughness of leaf surfaces and density of particles showed a close correlation. In general, the larger micro-roughness of leaf surfaces is, the larger density of particles is. Particles settling upon leaf surfaces were dominantly PM30 （particulate matter less than 10 μm in aerodynamic diameter; 98.4%） and PM2.5 （particulate matter less than 2.5 μm in aerodynamic diameter; 64.2%） which were closely relative to human health. Constant elements of particles were C, O, K, Ca, Si, Al, Mg, Na, Fe, S, Cl and minerals with higher content were SiO2, CaCO3, CaMg（CO3）2, NaCI and 2CASO4. H2O, SiO2. CaCO3 and CaMg（CO3）2 mainly came from resuspension of road dust. 2CaSO4. H2O was produced by the reaction between CaCO3 derived from earth dust or industrial emission and SO2, H2SO4 or sulfate. NaCl was derived from sea salt.     

Microbial changes in rhizospheric soils contaminated with petroleum hydrocarbons after bioremediation

Effects of bioremediation on microbial communities jn soils contaminated with petroleum hydrocarbons are a scientific problem to be solved. Changes in dominate microbial species and the total amount of microorganisms including bacteria and fungi in rhizospheric soils after bioremediation were thus evaluated using field bioremediation experiments. The results showed that there were changed dominant microorganisms including ] ] bacterial strains which are mostly Gram positive bacteria and 6 fungal species which were identified. The total amount of microorganisms including bacteria and fungi increased after bioremediation of microbial agents combined with planting maize. On the contrary, fungi in rhizospheric soils were inhibited by adding microbial agents combined with planting soybean.     

Quantification of multi-antibiotic resistant opportunistic pathogenic bacteria in bioaerosols in and around a pharmaceutical wastewater treatment plant

Pharmaceutical wastewater treatment plants(WWTPs) are thought to be a "seedbed" and reservoirs for multi-antibiotic resistant pathogenic bacteria which can be transmitted to the air environment through aeration. We quantified airborne multi-antibiotic resistance in a full-scale plant to treat antibiotics-producing wastewater by collecting bioaerosol samples from December2014 to July 2015. Gram-negative opportunistic pathogenic bacteria(GNOPB) were isolated, and antibiotic susceptibility tests against 18 commonly used antibiotics, including 11 β-lactam antibiotics, 3 aminoglycosides, 2 fluoroquinolones, 1 furan and 1 sulfonamide, were conducted.More than 45% of airborne bacteria isolated from the pharmaceutical WWTP were resistant to three or more antibiotics, and some opportunistic pathogenic strains were resistant to 16 antibiotics, whereas 45.3% and 50.3% of the strains isolated from residential community and municipal WWTP showed resistance to three or more antibiotics. The calculation of the multiple antibiotic resistance(MAR) index demonstrated that the air environment in the pharmaceutical WWTP was highly impacted by antibiotic resistance, while the residential community and municipal WWTP was less impacted by antibiotic resistance. In addition, we determined that the dominant genera of opportunistic pathogenic bacteria isolated from all bioaerosol samples were Acinetobacter, Alcaligenes, Citrobacter, Enterobacter, Escherichia, Klebsiella, Pantoea, Pseudomonas and Sphingomonas. Collectively, these results indicate the proliferations and spread of antibiotic resistance through bioaerosols in WWTP treating cephalosporin-producing wastewater, which imposed a potential health risk for the staff and residents in the neighborhood, calling for administrative measures to minimize the air-transmission hazard.     

Surface properties of bacteria from activated sludge in relation to bioflocculation

Two bacterial stains were isolated from the activated sludge and identified as Leucobacter sp.and Alcaligenes faecalis by 16S rDNA sequencing.Pure cultures of these two strains,representing well or poorly settled bacteria,were used to investigate the mechanism of bioflocculation in activated sludge.Based on the analyses of the characteristics of cells hydrophobicity,ζ-potential,flocculation ability and extracellular polymeric substance (EPS) composition under different growth stages,it was found that the ratio of cell EPS protein had the highly influence on ζ-potential and hydrophobicity,which were important factors to bioflocculation.Cellulase and Proteinase K could destroy the extracellular biopolymer and resulted in a decrease in the hydrophobicity and ζ-potential.However,in our study,the flocculation characteristics exhibited differently in relation to cellulase and Proteinase K.Flocculation of cells treated with cellulase and Proteinase K decreased sharply,and then recovered quickly in cellulase treatment,while cells treated with Proteinase K showed no sign of recovery.This reveals that the presence of protein in extracellular biopolymer plays an important role to the bioflocculation of cells.     

Effects of simulated acid rain on saplings of Pinus massoniana and Cunninghamia lanceolata

The effects of simulated acid rain with pH values of 6.63 (control, 4.5, 3.0, and 2.0 on saplings of Pinus massoniana and Cunninghamia lanceolate, were studied. The results showed that the pH of C.lanceolate, leaf sap and soil decreased as the acidity of rainfall increased. The acid rain with very low pH had significant effects on the photosynthetic rates per plant, but not on that of the per unit weight of dry leaves. The respiration rates of the two species were stimulated. Root and leaf boimass, but not stem biomass, were also reduced tremendously during a seven months period.     

Assessing the impact of fungicide enostroburin application on bacterial community in wheat phyllosphere

Fungicides have been used extensively for controlling fungal pathogens of plants. However, little is known regarding the effects that fungicides upon the indigenous bacterial communities within the plant phyllosphere. The aims of this study were to assess the impact of fungicide enostroburin upon bacterial communities in wheat phyllosphere. Culture-independent methodologies of 16S rDNA clone library and 16S rDNA directed polymerase chain reaction with denaturing gradient gel electrophoresis (PCR-DGGE) were used for monitoring the change of bacterial community. The 16S rDNA clone library and PCR-DGGE analysis both confirmed the microbial community of wheat plant phyllosphere were predominantly of the γ-Proteobacteria phyla. Results from PCR-DGGE analysis indicated a significant change in bacterial community structure within the phyllosphere following fungicide enostroburin application. Bands sequenced within control cultures were predominantly of Pseudomonas genus, but those bands sequenced in the treated samples were predominantly strains of Pantoea genus and Pseudomonas genus. Of interest was the appearance of two DGGE bands following fungicide treatment, one of which had sequence similarities (98%) to Pantoea sp. which might be a competitor of plant pathogens. This study revealed the wheat phyllosphere bacterial community composition and a shift in the bacterial community following fungicide enostroburin application.     

Isolation and characterization of facultative mixotrophic ammonia-oxidizing bacteria from constructed wetlands

Autotrophic ammonia-oxidizing bacteria (AOB) have been widely studied in constructed wetlands systems, while mixotrophic AOB have been less thoroughly examined. Heterotrophic bacteria were isolated from wastewater and rhizospheres of macrophytes of constructed wetlands, and then cultivated in a mixotrophic medium containing ammonium and acetic acid. A molecular characterization was accomplished using ITS-PCR amplification, and phylogenetic analysis based on 16S rRNA gene sequences. Results showed the presence of 35 bacteria, among 400 initially heterotrophic isolates, that were able to remove ammonia. These 35 isolates were classified into 10 genetically different groups based on ITS pattern. Then, a collection of 10 isolates were selected because of their relatively high ammonia removal efficiencies (ARE ≥ 80%) and their phylogenetic diversity. In conditions of mixotrophy, these strains were shown to be able to grow (increase of optical density OD₆₆₀ during incubation with assimilation of nitrogen into cellular biomass) and to oxidize ammonia (important ammonia oxidation efficiencies, AOE between 79% and 87%). Among these facultative mixotrophic AOB, four isolates were genetically related to Firmicutes (Bacillus and Exiguobacterium), three isolates were affiliated to Actinobacteria (Arthrobacter) and three other isolates were associated with Proteobacteria (Pseudomonas, Ochrobactrum and Bordetella).     

Monitoring impact of mefenacet treatment on soil microbial communities by PCR-DGGE fingerprinting and conventional testing procedures

The effect of acetanilide herbicide mefenacet on soil microbial communities was studied using paddy soil samples with different short-term treatments. The culturable bacteria （plate counts）, dehydrogenase activity and changes in community structure （denaturing gradient gel electrophoresis （DGGE） analysis） were used for biological community assessments. Mefenacet was a significant stimulus to cultural aerobic bacteria and dehydrogenase activity while Sphingobacterium multivorum Y1, a bacterium efficiently degrading the mefenacet, only induced the increasing colony-forming unit （CFU） of bacteria but little effect on dehydrogenase activity during the whole experiment. The degree of similarity between the 16S rDNA profiles of the communities was quantified by numerically analyzing the DGGE band patterns. Similarity dendrograms showed that the microbial community structures of the mefenacet-treated and non-treated soils were not significantly different. But supplement of S. multivorum Y1 could increase the diversity of the microbial community in the mefenacet-polluted paddy soil. This work is a new attempt to apply the S. multivorum Y1 for remediation of the mefenacet-polluted environments.     

A preliminary analysis of the surface chemistry of atmospheric aerosol particles in a typical urban area of Beijing

Atmospheric aerosol particle samples were collected using an Ambient Eight Stage(Non-Viable) Cascade Impactor Sampler in a typical urban area of Beijing from 27 th Sep.to 5th Oct.,2009.The surface chemistry of these aerosol particles was analyzed using Static Time of Flight-Secondary Ion Mass Spectrometry(Static TOF-SIMS).The factors influencing surface compositions were evaluated in conjunction with the air pollution levels,meteorological factors,and air mass transport for the sampling period.The results show that a variety of organic ion groups and inorganic ions/ion groups were accumulated on the surfaces of aerosol particles in urban areas of Beijing;and hydrophobic organic compounds with short-or middle-chain alkyl as well as hydrophilic secondary inorganic compounds were observed.All these compounds have the potential to affect the atmospheric behavior of urban aerosol particles.PM1.1–2.1and PM3.3–4.7had similar elements on their surfaces,but some molecules and ionic groups demonstrated differences in Time of Flight-Secondary Ion Mass Spectrometry spectra.This suggests that the quantities of elements varied between PM1.1–2.1and PM3.3–4.7.In particular,more intense research efforts into fluoride pollution are required,because the fluorides on aerosol surfaces have the potential to harm human health.The levels of air pollution had the most significant influence on the surface compositions of aerosol particles in our study.Hence,heavier air pollution was associated with more complex surface compositions on aerosol particles.In addition,wind,rainfall,and air masses from the south also greatly influenced the surface compositions of these urban aerosol particles.     

Removal of airborne microorganisms emitted from a wastewater treatment oxidation ditch by adsorption on activated carbon

Bioaerosol emissions from wastewater and wastewater treatment processes are a significant subgroup of atmospheric aerosols. Most previous work has focused on the evaluation of their biological risks. In this study, however, the adsorption method was applied to reduce airborne microorganisms generated from a pilot scale wastewater treatment facility with oxidation ditch. Results showed adsorption on granule activated carbon (GAC) was an e cient method for the purification of airborne microorganisms. The GAC itself had a maximum adsorption capacity of 2217 CFU/g for airborne bacteria and 225 CFU/g for fungi with a flow rate of 1.50 m3/hr. Over 85% of airborne bacteria and fungi emitted from the oxidation ditch were adsorbed within 80 hr of continuous operation mode. Most of them had a particle size of 0.65–4.7 m. Those airborne microorganisms with small particle size were apt to be adsorbed. The SEM/EDAX, BET and Boehm’s titration methods were applied to analyse the physicochemical characteristics of the GAC. Relationships between GAC surface characteristics and its adsorption performance demonstrated that porous structure, large surface area, and hydrophobicity rendered GAC an e ective absorber of airborne microorganisms. Two regenerate methods, ultraviolet irradiation and high pressure vapor, were compared for the regeneration of used activated carbon. High pressure vapor was an e ective technique as it totally destroyed the microorganisms adhered to the activated carbon. Microscopic observation was also carried out to investigate original and used adsorbents.     

Effects of used lubricating oil on two mangroves Aegiceras corniculatum and Avicennia marina

An outdoor experiment was set up to investigate the effects of used lubricating oil(5 L/m~2)on Aegiceras corniculatum Blanco. and Avicennia marina(Forsk)Vierh.,two salt-excreting mangroves.A.marina was more sensitive to used lubricating oil than A. corniculatum and canopy-oiling resulted in more direct physical damage and stronger lethal effects than base-oiling.When treated with canopy-oiling,half of A.corniculatum plants survived for the whole treatment time(90 d);but,for A.marina,high mortality (83%)resulted from canopy-oiling within 3 weeks and no plants survived for 80 d.Base-oiling had no lethal effects on A.corniculatum plants even at the termination of this experiment,but 83% of A.marina plants died 80 d after treatment.Forty days after canopy- oiling,93% ofA.corniculatum leaves fell and no live leaves remained on A.marina plants.By the end of the experiment,base-oiling treatment resulted in about 45% ofA.corniculatum leaves falling,while all A.marina leaves and buds were burned to die.Lubricating oil resulted in physiological damage to A.corniculatum leaves,including decreases in chlorophyll and carotenoid contents,nitrate reductase,peroxidase and superoxide dismutase activities,and increases in malonaldehyde contents.For both species,oil pollution significantly reduced leaf,root,and total biomass,but did not significantly affect stem biomass.Oil pollution resulted in damage to the xylem vessels of fine roots but not to those of mediate roots.     

Morphological and semi-quantitative characteristics of diesel soot agglomerates emitted from commercial vehicles and a dynamometer

Diesel soot aggregates emitted from a model dynamometer and 11 on-road vehicles were segregated by a micro-orifice uniform deposit impactor (MOUDI). The elemental contents and morphological parameters of the aggregates were then examined by scanning electron microscopy coupled with an energy dispersive spectrometer (SEM-EDS), and combined with a fractional Brownian motion (fBm) processor. Two mode-size distributions of aggregates collected from diesel vehicles were confirmed. Mean mass concentration of 339 mg/m3 (dC/dlogdp) existed in the dominant mode (180–320 nm). A relatively high proportion of these aggregates appeared in PM1, accentuating the relevance regarding adverse health e ects. Furthermore, the fBm processor directly parameterized the SEM images of fractal like aggregates and successfully quantified surface texture to extract Hurst coe cients (H) of the aggregates. For aggregates from vehicles equipped with a universal cylinder number, the H value was independent of engine operational conditions. A small H value existed in emitted aggregates from vehicles with a large number of cylinders. This study found that aggregate fractal dimension related to H was in the range of 1.641–1.775, which is in agreement with values reported by previous TEM-based experiments. According to EDS analysis, carbon content ranged in a high level of 30%–50% by weight for diesel soot aggregates. The presence of Na and Mg elements in these sampled aggregates indicated the likelihood that some engine enhancers composed of biofuel or surfactants were commonly used in on-road vehicles in Taiwan. In particular, the morphological H combined with carbon content detection can be useful for characterizing chain-like or cluster diesel soot aggregates in the atmosphere.