Continuous-flow Chamber Research Articles

A custom, low-cost, continuous flow chamber built for experimental Sargassum seaweed decomposition and exposure of small rodents to generated gaseous products

Since 2011, Sargassum events have increased in frequency along the Caribbean and Atlantic coasts. The accumulation and decomposition of large amounts of Sargassum seaweed on beaches pose socio-economic, ecological, and health risks due to the emission of hydrogen sulfide (H2S), methane, and ammonia. However, limited research exists on the emission processes and the health effects of subchronic and chronic exposure to low levels of H2S. Additionally, the absence of emission factor data for Sargassum decomposition on-site makes health risk assessments challenging. This study aimed to create a custom chamber to simulate real-world Sargassum decomposition, exposing experimental animals to the generated gases. Metal content was analyzed, and emission rates were estimated in a controlled environment. The decomposition-exposure system replicated reported environmental gas emissions from the Caribbean region, except for NH3. H2S bursts were observed during the decomposition process at intervals of 2–10 days, with higher frequency associated with larger masses of decomposing Sargassum. The decomposed gas was transferred to the exposure chamber, resulting in an 80–87% reduction in H2S concentration. The maximum H2S emission was 156 ppm, with a concentration ranging from 50.4 to 56.5 ppm. An estimated emission rate of 7–8 g/h for H2S was observed, and significant levels of lead, arsenic, and aluminum were found in beached Sargassum from the northeast coast of Brazil. This study's developed model provides an opportunity to investigate the effects and risks to human health associated with exposure to gases produced during the environmental decomposition of Sargassum seaweed.

Description of a Dimethylmercury Automatic Analyzer for the High-Resolution Measurement of Dissolved Gaseous Mercury Species in Surface Ocean Waters.

Our understanding of the significance of dimethylmercury (DMHg) to the mercury (Hg) global ocean biogeochemical cycle is unclear because of the lack of detailed DMHg measurements in the water column. To our knowledge, 30 years of published studies have generated no more than 200 DMHg data points in the ocean surface waters and marine boundary layer (MBL). To improve the precision and reduce the uncertainty in determining DMHg in surface seawater, we developed a simple and robust DMHg automatic analyzer (DAA). This DAA system couples the main sampling and analytic steps, including a continuous flow chamber, with dual Carbotrap preconcentration, a gas chromatographic column, a cold vapor atomic fluorescence spectrometry, and a data logger for signal integration. We compared the operation, performance, and reproducibility between our DAA and the traditional manual analytic method. Its advantages include the ease of operation, the high time resolution and precision (30 min sampling and <5% relative variation), and long-term stability (2 weeks). The DAA can determine DMHg in both the MBL and surface seawater. The estimated detection limits for DMHg with the DAA in the atmosphere and in surface seawater are 10 pg/m3 and 0.2 fM, respectively. The successful DAA field measurement in coastal waters indicates that it can help detect the low DMHg concentration in surface seawater, and the time series DMHg data helped our understanding of the DMHg behavior (sources and sinks) and its flux into the MBL. The comparison of DMHg concentration in various oceans also suggests that the coastal region had the lowest averaged DMHg, up to an order of magnitude lower than other ecosystems.

Monoterpene Photooxidation in a Continuous-Flow Chamber: SOA Yields and Impacts of Oxidants, NOx, and VOC Precursors.

Monoterpene photooxidation plays an important role in secondary organic aerosol (SOA) formation in the atmosphere. The low-volatility products can enhance new particle formation and particle growth and thus influence climate feedback. Here, we present the results of α-pinene and Δ-3-carene photooxidation experiments conducted in continuous-flow mode in an environmental chamber under several reaction conditions. The roles of oxidants, addition of NO, and VOC molecular structure in influencing SOA yield are illustrated. SOA yield from α-pinene photooxidation shows a weak dependence on H2O2 concentration, which is a proxy for HO2 concentration. The high O/C ratios observed in the α-pinene photooxidation products suggest the production of highly oxygenated organic molecules (HOM). Addition of ozone to the chamber during low-NOx photooxidation experiments leads to higher SOA yield. With the addition of NO, the production of N-containing HOMs is enhanced and the SOA yield shows a modest, nonlinear dependence on the input NO concentration. Carene photooxidation leads to higher SOA yield than α-pinene under similar reaction conditions, which agrees with the lower volatility retrieved from evaporation kinetics experiments. These results improve the understanding of SOA formation from monoterpene photooxidation and could be applied to refine the representation of biogenic SOA formation in models.

Particle emissions from a modern heavy-duty diesel engine as ice nuclei in immersion freezing mode: a laboratory study on fossil and renewable fuels

Abstract. We studied ice-nucleating abilities of particulate emissions from a modern heavy-duty diesel engine using three different types of fuel. The polydisperse particle emissions were sampled during engine operation and introduced to a continuous-flow diffusion chamber (CFDC) instrument at a constant relative humidity RHwater=110 %, while the temperature was ramped between −43 and −32 ∘C (T scan). The tested fuels were EN 590 compliant low-sulfur fossil diesel, hydrotreated vegetable oil (HVO), and rapeseed methyl ester (RME); all were tested without blending. Sampling was carried out at different stages in the engine exhaust aftertreatment system, with and without simulated atmospheric processing using an oxidation flow reactor. In addition to ice nucleation experiments, we used supportive instrumentation to characterize the emitted particles for their physicochemical properties and presented six parameters. We found that the studied emissions contained no significant concentrations of ice-nucleating particles likely to be of atmospheric relevance. The substitution of fossil diesel with renewable fuels, using different emission aftertreatment systems such as a diesel oxidation catalyst, and photochemical aging of total exhaust had only minor effect on their ice-nucleating abilities.

Assessment of Daytime HONO Emission Source from Asphalt Surface to Urban Air

Existing studies suggest various potential daytime sources of atmospheric nitrous acid (HONO), including photolysis surface reactions and photo-enhanced NO2 conversion on organic surfaces. However, the understanding of daytime HONO sources is still inadequate. In this study, we report the HONO formation on asphalt surfaces under various NO2, VOCs (toluene and hexane), and UV irradiance conditions using a continuous flow chamber. Although no HONO formation was found without light exposure, the light threshold for HONO formation on the asphalt surface was very low, with a total UV (TUV) of 0.7 W m−2. HONO formation on the asphalt surface was linearly dependent on NO2 up to 300 ppb in the presence of VOCs, but no HONO formation was observed with humified air and NO2. HONO production was saturated at high hydrocarbon concentrations and light intensities. The calculated first-order NO2 conversion rate to HONO on the asphalt surface was 1.2 × 10−4 s −1. The observed mean HONO emission flux was 1.3 × 109 molecules cm−2 s −1 with a similar range of those on other urban covered surfaces. The calculated vertical HONO profile using the measured HONO emission flux and 1-D steady state model revealed that the asphalt surface may account for 13% of daytime HONO in the elevated on-road pollutant concentrations in Seoul. However, we show that its HONO contribution could be much higher on real-life road surfaces directly exposed to much higher NO2 emissions from vehicle exhaust.

Always Lost but Never Forgotten: Gas-Phase Wall Losses Are Important in All Teflon Environmental Chambers.

Yields of secondary organic aerosol (SOA) formation from oxidation of volatile organic compounds are measured in laboratory chambers and then applied in regional and global models. Gas-phase losses to large Teflon-walled environmental chambers have been recently shown to reduce SOA yields. Historically, most chambers have operated in batch mode. Increasingly, however, continuous flow (CF) mode is being used, in which reactants and products are continuously introduced and exhausted from the chamber. Recent literature reports indicate a belief that SOA yields measured in CF chambers are not affected by gas-phase wall losses (GWL). Here, we use an experimentally-constrained box model to show that gas-phase wall losses impact both types of chambers when run under similar conditions. We find CF experiments do mitigate some effects of gas-phase wall losses after long (>2 days) experiment run times, but they have significant losses for typical literature experiment times of 1 day. However, this mitigation phenomenon is an experiment- and mechanism-dependent, and GWL still affects the absolute SOA yield. Finally, we show that at condensation sink values higher than the wall loss rate a lack of change in yield vs seed surface area does not necessarily indicate whether GWL affects the experiment and does not suggest the magnitude.

Experimental investigation into the volatilities of highly oxygenated organic molecules (HOMs)

Abstract. Secondary organic aerosol (SOA) forms a major part of the tropospheric submicron aerosol. Still, the exact formation mechanisms of SOA have remained elusive. Recently, a newly discovered group of oxidation products of volatile organic compounds (VOCs), highly oxygenated organic molecules (HOMs), have been proposed to be responsible for a large fraction of SOA formation. To assess the potential of HOMs to form SOA and to even take part in new particle formation, knowledge of their exact volatilities is essential. However, due to their exotic, and partially unknown, structures, estimating their volatility is challenging. In this study, we performed a set of continuous flow chamber experiments, supported by box modelling, to study the volatilities of HOMs, along with some less oxygenated compounds, formed in the ozonolysis of α-pinene, an abundant VOC emitted by boreal forests. Along with gaseous precursors, we periodically injected inorganic seed aerosol into the chamber to vary the condensation sink (CS) of low-volatility vapours. We monitored the decrease of oxidation products in the gas phase in response to increasing CS, and were able to relate the responses to the volatilities of the compounds. We found that HOM monomers are mainly of low volatility, with a small fraction being semi-volatile. HOM dimers were all at least low volatility, but probably extremely low volatility; however, our method is not directly able to distinguish between the two. We were able to model the volatility of the oxidation products in terms of their carbon, hydrogen, oxygen and nitrogen numbers. We found that increasing levels of oxygenation correspond to lower volatilities, as expected, but that the decrease is less steep than would be expected based on many existing models for volatility, such as SIMPOL. The hydrogen number of a compound also predicted its volatility, independently of the carbon number, with higher hydrogen numbers corresponding to lower volatilities. This can be explained in terms of the functional groups making up a molecule: high hydrogen numbers are associated with, e.g. hydroxy groups, which lower volatility more than, e.g. carbonyls, which are associated with a lower hydrogen number. The method presented should be applicable to systems other than α-pinene ozonolysis, and with different organic loadings, in order to study different volatility ranges.

Biofilm-Forming by Carbapenem Resistant Enterobacteriaceae May Contribute to the Blood Stream Infection.

Bloodstream infection (BSI) due to carbapenem-resistant Enterobacteriaceae (CRE) has a high mortality rate and is a serious threat worldwide. Ten CRE strains (eight Enterobacter cloacae, one Klebsiella pneumoniae and one Citrobacter freundii) were isolated from the blood of nine patients, a percentage of whom had been treated with indwelling devices. The steps taken to establish cause included minimum inhibitory concentration (MIC) tests, a pulsed-field gel electrophoresis (PFGE), biofilm study, a multiplex PCR for resistant genes of carbapenemases and extended-spectrum beta-lactamases (ESBLs), and plasmid incompatibility typing. All strains showed a tendency toward resistance to multiple antibiotics, including carbapenems. Frequently isolated genes of ESBLs and carbapenemases include blaTEM-1 (four strains), blaSHV-12 (four strains) and blaIMP-1 (six strains). A molecular analysis by PFGE was used to divide the XbaI-digested genomic DNAs of 10 CRE strains into eight patterns, and the analysis showed that three E. cloacae strains detected from two patients were either identical or closely related. The biofilm production of all CRE strains was examined using a microtiter biofilm assay, and biofilm growth in continuous flow chambers was observed via the use of a confocal laser scanning microscope. Our study indicates that biofilm formation on indwelling devices may pose a risk of BSI due to CRE.

A steady-state continuous flow chamber for the study of daytime and nighttime chemistry under atmospherically relevant NO levels

Abstract. Experiments performed in laboratory chambers have contributed significantly to the understanding of the fundamental kinetics and mechanisms of the chemical reactions occurring in the atmosphere. Two chemical regimes, classified as “high-NO” vs. “zero-NO” conditions, have been extensively studied in previous chamber experiments. Results derived from these two chemical scenarios are widely parameterized in chemical transport models to represent key atmospheric processes in urban and pristine environments. As the anthropogenic NOx emissions in the United States have decreased remarkably in the past few decades, the classic “high-NO” and “zero-NO” conditions are no longer applicable to many regions that are constantly impacted by both polluted and background air masses. We present here the development and characterization of the NCAR Atmospheric Simulation Chamber, which is operated in steady-state continuous flow mode for the study of atmospheric chemistry under “intermediate NO” conditions. This particular chemical regime is characterized by constant sub-ppb levels of NO and can be created in the chamber by precise control of the inflow NO concentration and the ratio of chamber mixing to residence timescales. Over the range of conditions achievable in the chamber, the lifetime of peroxy radicals (RO2), a key intermediate from the atmospheric degradation of volatile organic compounds (VOCs), can be extended to several minutes, and a diverse array of reaction pathways, including unimolecular pathways and bimolecular reactions with NO and HO2, can thus be explored. Characterization experiments under photolytic and dark conditions were performed and, in conjunction with model predictions, provide a basis for interpretation of prevailing atmospheric processes in environments with intertwined biogenic and anthropogenic activities. We demonstrate the proof of concept of the steady-state continuous flow chamber operation through measurements of major first-generation products, methacrolein (MACR) and methyl vinyl ketone (MVK), from OH- and NO3-initiated oxidation of isoprene.

Ultrasound-assisted crystallization of lactose in the presence of whey proteins and κ-carrageenan

The conventional process of lactose crystallization is prolonged, hardly controllable and the crystals have low quality. In this work, the effect of ultrasound on the crystallization of lactose in an aqueous system was assessed. Additionally, it was studied how the presence of whey proteins (which are a common impurity) and κ-carrageenan (that possess high water-binding capacity) could modify the process of lactose crystallization. Lactose solutions at 25% were sonicated in a continuous flow chamber at two different energy densities (9 and 50 J mL−1) before the start of crystallization. Some of these lactose solutions were previously added with κ-carrageenan (0, 150 and 300 mg L−1), with whey proteins (0.64%) or with both at the same time. Ultrasound sped up the rate of crystallization, decreased the crystal’s size and narrowed the crystal size distribution (CSD). The presence of whey proteins accelerated the process of crystallization but induced the formation of amorphous lactose. Likewise, the rate of lactose crystallization was improved by the addition of 150 mg L−1 of carrageenan. Whereas, the combination of carrageenan and whey proteins generated the smallest crystals (6 μm), the narrowest CSD and minimized the formation of amorphous lactose.

Aminobacter sp. MSH1 invades sand filter community biofilms while retaining 2,6-dichlorobenzamide degradation functionality under C- and N-limiting conditions.

Aminobacter sp. MSH1 is of interest for bioaugmentation of biofiltration units in drinking water treatment plants (DWTPs) due to its ability to degrade the groundwater micropollutant 2,6-dichlorobenzamide (BAM). Using a continuous flow chamber biofilm model, MSH1 was previously shown to colonize surfaces and degrade BAM at trace concentrations as low as 1 μg/L under the oligotrophic conditions found in DWTPs. In DWTP filtration units, MSH1 has to compete with the resident biofilm microbiota for space and nutrients. Using the same model, we examined how a sand filter community (SFC) affects MSH1's BAM-degrading activity and biofilm formation under C- and N-limiting conditions when fed with trace concentrations of BAM. MSH1 was inoculated simultaneously with the SFC (co-colonization mode) or after the SFC formed a biofilm (invasion mode). MSH1 successfully established in the SFC biofilm showing growth and activity. In co-colonization mode, MSH1 decreased in number in the presence of the SFC and formed isolated colonies, while specific BAM-degradation activity increased. In the invasion mode, MSH1 also decreased in numbers in the presence of the SFC but formed mixed colonies, while specific BAM degradation was unaffected. Our results show that MSH1 invades and performs successfully in an SFC biofilm under the oligotrophic conditions of DWTPs.

Quantification of Low-Level Drug Effects Using Real-Time,in vitroMeasurement of Oxygen Consumption Rate

There is a general need to detect toxic effects of drugs during preclinical screening. We propose that increased sensitivity of xenobiotics toxicity combined with improved in vitro physiological recapitulation will more accurately assess potentially toxic perturbations of cellular biochemistry that are near in vivo pharmacological exposure levels. Importantly, measurement of such cytopathologies avoids activating mechanisms mediating toxicity at suprapharmacologic levels not relevant to in vivo effects. We present a sensitive method to measure changes in oxygen consumption rate (OCR), a well-established parameter reflecting a potential hazard, in response to exposure to pharmacologic levels of drugs using a flow culture system and state of the art oxygen sensing system. We tested metformin and acetaminophen on rat liver slices to illustrate the method. The features of the method include continuous and very stable measurement of OCR over the course of 48 h in liver slices in a continuous flow chamber with the ability to resolve changes as small as 0.3%/h. Kinetic modeling of metformin inhibition of OCR over a wide range of concentrations revealed both a slow and fast mechanism, where the fast mechanism activated only at concentrations above 0.6 mM. For both drugs, small amounts of inhibition were reversible, but higher decrements were irreversible. Overall the study highlights the advantages of measuring low-level toxicity so as to avoid the common extrapolations made about drug toxicity based on effects of drugs tested at suprapharmacologic levels.

Elution of High Dose Amphotericin B Deoxycholate From Polymethylmethacrylate

Fungal periprosthetic joint infections are rare, devastating complications of arthroplasty. There is conflicting evidence as to the efficacy of amphotericin B elution from cement spacers. The purpose of this study was to determine whether concentrations of amphotericin B released from bone cement over time would be efficacious in treating a periprosthetic infection. A continuous flow chamber was used to evaluate the in vitro release of amphotericin from cement beads containing 7.5% amphotericin. Following polymerization, 3.3% of the initially loaded amphotericin B was detected. The peak mean concentration eluted from the bone cement was 0.33 μg/mL at 8 hours. The AUC0–24 was 2.79 μg/mL/h; 0.20% of the amphotericin B was released. In conclusion, amphotericin B is released from bone cement at a clinically useful concentration.

Factors responsible for spatial and temporal variation of soil CO2 efflux in a 50 year recovering tropical forest, Peninsular Malaysia

Environmental abiotic and biotic factors are important in controlling soil CO2 efflux in forest ecosystems of different ages, as they play an important role in soil respiration. In understanding the spatial and temporal variation of soil CO2 efflux after several years of forest logging, there is a need to quantify the changing soil properties, environmental factors, and the total above and below ground biomass. This study was conducted in a 50-year old recovering tropical lowland forest in Peninsular Malaysia, measuring soil CO2 efflux using the continuous open flow chambers technique connected to a multi gas-handling unit and infrared gas analyser. The aim of this study was to determine the spatial and temporal variation of soil CO2 efflux in relation to changes in soil properties, environmental factors and forest carbon in a recovering forest. The efflux rates of about 389.20, 634.78, 564.81, 537.92 and 428.72 mg m−2 h−1, respectively, varied across the days and months, increasing from February and attaining the maximum in March and then gradually decreasing from April to June. The soil properties revealed a considerable amount of soil organic carbon, total organic carbon, and soil organic carbon stock, while the total above ground biomass, below ground biomass, soil pH, nitrogen to carbon ratio were found to provide nutrients for microbial activities in soil and to emit soil CO2. The multiple linear regression model indicated that the soil temperature and moisture explained the spatial and temporal variation in soil CO2 efflux; likewise, the changes in the soil properties and forest carbon significantly increased the soil CO2 efflux indicating a strong positive correlation (R 2 = 0.93).

Eastern oyster (Crassostrea virginica) filtration, biodeposition, and sediment nitrogen cycling at two oyster reefs with contrasting water quality in Great Bay Estuary (New Hampshire, USA)

Benthic deposition of carbon (C) and nitrogen (N)-rich oyster biodeposits may increase denitrification, or anaerobic respiration of nitrate (NO3 −) to di-nitrogen gas (N2). However, environmental drivers of C and N dynamics in oyster biodeposits and reef-adjacent sediments require clarification. In July 2012, we collected intact sediment cores adjacent to and 15–20 m away from two oyster reefs (Crassostrea virginica) in Great Bay, New Hampshire, USA: one reference site and one site with cultural eutrophication. We also measured seston, chlorophyll a, and in situ oyster feeding and biodeposition. Cores were incubated in continuous-flow chambers where inflow water received 15N-ammonium (NH4 +), 15NO3 −, or no isotopes (control). We quantified fluxes of dissolved nutrients and gasses (oxygen, 28N2, 29N2, 30N2, and argon) after 24 h. Finally, we measured size-fractionated sediment organic matter. At the eutrophic site, abundant phytoplankton in the 5–28 µm size range was correlated with enhanced oyster feeding rates and biodeposit quality (lower C:N). This site had greater denitrification rates in reef-adjacent cores relative to distal cores. Low production of 29,30N2 in 15NH4 + amended cores suggested water column or biodeposit NH4 + were unlikely to be converted to N2. At both sites, reef-adjacent cores had more shell and higher 29,30N2 production with 15NO3 − addition relative to distal cores, suggesting direct denitrification enhancement near reefs. Oysters likely increased sediment N2 production via high quality biodeposits (eutrophic site), and NO3 − diffusion via structural complexity of reef-adjacent sediment (both sites). Overall, results suggest oyster-mediated ecosystems services may be expected to vary with environmental conditions.

Biofilm formation of a bacterial consortium on linuron at micropollutant concentrations in continuous flow chambers and the impact of dissolved organic matter

Bacterial multispecies biofilms are catalysts for pollutant degradation in aqueous ecosystems. Their activity in systems where xenobiotics occur as micropollutants (μg L(-1) level) and natural dissolved organic matter provides carbon and energy instead remains uncharacterized. Biofilm formation of a bacterial consortium consisting of the linuron-degrading Variovorax sp. WDL1 and metabolite-degrading strains Comamonas sp. WDL7 and Hyphomicrobium sp. WDL6 at micropollutant linuron concentrations and the impact of auxiliary carbon sources on degradation and biofilm composition were investigated. Biofilms formed at concentrations of 1000, 100, and 10 μg L(-1) linuron. The highest biomass, organized in mixed-species mounds, was observed at 1000 μg L(-1) linuron, while at 100 and 10 μg L(-1) , thin layers of cells occurred. Linuron removal efficiencies decreased from c. 85% when fed with 100 and 1000 μg L(-1) linuron to 30% in case of 10 μg L(-1) linuron due to reduced specific activity. Biofilms grown on 10 μg L(-1) linuron were subsequently fed with easily and less degradable carbon sources in addition to 10 μg L(-1) linuron. Although co-feeding with more degradable C-sources increased biofilm biomass, linuron removal remained 30%. Calculations based on biofilm volume measurements pointed toward reduced specific activity, compensated by a higher biomass. Uncertainties about biofilm heterogeneity and cell volume can undo this explanation.

Forest Recovering and Soil Respiration Rate

This study was conducted to investigate the rate of soil respiration from a recovering forest of the tropics and its relationship with changes in environmental factorsafter years of deforestation. Soil respiration measurement was conducted using the continuous open flow chamber technique connected to a multi gas- handling unit and infrared gas analyser, while the forest biomass and soil properties were quantified using the Kjeldahl method and Walkley-black wet oxidation technique. The average means soil respiration rate were 341.23, 383.07, 340.30, 308.12, 286.07, 256.05 mg m -2 h -1 between June and December. Soil respiration in the month of July was significantly (p<0.01) higher compare to other months, with lower emission rate in December. Soil respiration exhibited a variation pattern that was similar to soil temperature pattern, the pattern varied monthly. Likewise, the forest carbon input and soil properties were found to be significantly correlated with soil respiration as they provide nutrients for microorganism to emit soil CO2. The remarkable soil CO2 emission from the recovering forest was attributed to changes in environmental factors as the forest is recovering from deforestation. The correlation and multiple linear regression model proved that environmental factors influenced the high rate of soil CO2 emission indicating a strong positive relationship (0.94; p<0.01). These results suggestthat forest recovering could still emit considerable percentages of soil CO2 due to the impact from deforestation which could have a great implication on environmental factors and the atmospheric carbon balance.

Forest Logging and It Impact on Soil Carbon Dioxide Effluxin the tropical Forest, Peninsular Malaysia

Forest harvesting is expected to have an impact on soil CO2 efflux as it influence soil properties and changes in microclimatic conditions which can have implications on the regional carbon balance. Soil CO2 efflux was measured using a continuous open flow chambers technique connected to a multi-gas-handling unit and infrared CO2/H2O gas analyser. Soil temperature, soil moisture, water potential, Total Organic Carbon (TOC), Soil Organic Carbon (SOC), Soil Organic Carbon stock (SOCstock), Bulk density and pH were examinedto ascertain their contribution onsoil CO2 efflux and effect ofenvironmental factors in a canopy gap created through the logging of groups of trees in the Sungai Menyala forest, Peninsular Malaysia.

Crop optimization and pre-steps standardization to get a Bipolaris euphorbiae-based bioherbicide

Making bioproducts available to the market requires finding appropriate processes for mass production and formulation of biological agents. This study aimed at evaluating the Bipolaris euphorbiae production in a solid medium (fermentation in solid substrate) and in a biphasic system (growth in a liquid medium followed by growth in a solid medium), as well as determining the processes for collecting and drying conidia, under laboratory conditions. The influence of the incubation period and inoculum quantity were also investigated. The conidia were dried by using an oven (30ºC, 35ºC, 40ºC, 45ºC, 50ºC, 55ºC and 60ºC), and laminar flow, continuous air flow and aseptic chamber at room temperature. Dry conidia were obtained by sieving and grinding in a ball mill, hammer mill or grain grinder. The conidia viability and sporulation efficiency were evaluated in the solid medium and in the biphasic system. For growth period, the best sporulation on solid medium was obtained after 10 days of incubation, reaching 8.3 x 10(7) conidia g-1 of substrate. The biphasic system did not increase the B. euphorbiae sporulation (4.5 x 10(7) conidia g-1 of substrate), after 14 days, and the amount of liquid inoculum used in this system was not an important factor for increasing its production. The continuous air flow and laminar flow preserved the conidial viability (94.6% and 99.1%, respectively), while promoting a great moisture loss (62.6% and 54.0%, respectively). All the grinding processes reduced the conidia germination (86.2%, 10.5% and 12%, respectively), while sieving allowed the collecting of powdered conidia with high viability (94.8%).

The relationship between biofilm formations and capsule in Haemophilus influenzae

To evaluate the biofilm formation of non-typeable Haemophilus influenzae (NTHi) and H. influenzae type b (Hib) clinical isolates, we conducted the following study. Serotyping and polymerase chain reaction were performed to identify β-lactamase-negative ampicillin (ABPC)-susceptible (BLNAS), β-lactamase-negative ABPC-resistant (BLNAR), TEM-1 type β-lactamase-producing ABPC-resistant (BLPAR)-NTHi, and Hib. Biofilm formation was investigated by microtiter biofilm assay, as well as visually observation with a scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) in a continuous-flow chamber. As a result, totally 99 strains were investigated, and were classified into 4 groups which were 26 gBLNAS, 22 gBLNAR, 28 gBLPAR-NTHi and 23 Hib strains. The mean OD600 in the microtiter biofilm assay of gBLNAS, gBLNAR, gBLPAR-NTHi, and Hib strains were 0.57, 0.50, 0.34, and 0.08, respectively. NTHi strains were similar in terms of biofilm formations, which were observed by SEM and CLSM. Five Hib strains with the alternated type b cap loci showed significantly increased biofilm production than the other Hib strains. In conclusion, gBLNAS, gBLNAR, and gBLPAR-NTHi strains were more capable to produce biofilms compared to Hib strains. Our data suggested that resistant status may not be a key factor but capsule seemed to play an important role in H. influenzae biofilm formation.