Quantification of airborne microorganisms and investigation of their interactions with non-living particles

A fluorescence method was developed for the direct counting of airborne microorganisms and for the examination of their interactions with other aerosol particles. The method is based on a combination of the aerosol sampling technique using a cascade impactor and the selective dyeing of the trapped microorganisms with ethidium bromide. The method enables both the total microorganism concentrations and their counts in clusters to be evaluated. The new method and the cultivation method, enabling the colony-forming units (CFU) to be determined, were compared. Based on the results obtained by the fluorescence technique, a procedure is suggested for the conversion of CFU data to bacteria and yeast concentrations.     

Depth profiling of the elemental surface composition of the oral microorganism S. salivarius HB and fibrillar mutants by X-ray photoelectron spectroscopy.

X-ray photoelectron spectroscopy (XPS) on microbial cell surfaces requires freeze-drying of cells, and as a result, the cell surface appendages flatten out on the cell surface and form a collapsed fibrillar mass. At present, it is unclear how the density, length and composition of these fibrils influence the elemental surface composition as probed by XPS. The sampling depth of XPS can be varied by changing the electron take-off angle. In this article, we made a depth profiling of the collapsed fibrillar mass of Streptococcus salivarius HB and fibril-deficient mutants by angle-dependent XPS. Methylamine tungstate negative staining and ruthenium red staining followed by sectioning revealed distinct classes of fibrils with various lengths on each of the strains. Interpretation of the angle dependence of the oxygen/carbon (O/C) and phosphorus/carbon (P/C) surface concentration ratios of these strains was difficult. However, the angle dependence of the nitrogen/carbon (N/C) surface concentration ratio could be fully interpreted: N/C did not vary with sampling depth on a bald strain, S. salivarius HBC12 and on S. salivarius HB7, a strain with a dense array of fibrils of uniform length. N/C decreased with sampling depth in case of a sparsely fibrillated strain, S. salivarius HBV51 and eventually reached the value observed for the bald strain, HBC12. A high N/C at small sampling depth was observed for S. salivarius HB with protruding, protein rich fibrils. We conclude that elemental depth profiling of microbial cell surfaces by XPS can be interpreted to coincide with structural and biochemical information on the cell surface as obtained by electron microscopy and can therefore be considered as a useful technique to study structural features of cell surfaces in combination with electron microscopy.     

Evaluation of Abbreviated Impactor Measurements (AIM) and Efficient Data Analysis (EDA) for Dry Powder Inhalers (DPIs) Against the Full-Resolution Next Generation Impactor (NGI)

The full-resolution next generation impactor (NGI) and three abbreviated impactor systems were used to obtain the apparent aerodynamic particle size distribution (APSD) and other quality measures for marketed dry powder inhalers (DPIs) using the compendial method and efficient data analysis (EDA). APSD for the active pharmaceutical ingredient (API) in Spiriva^® Handihaler^®, Foradil^® Aerolizer^®, and Relenza^® Diskhaler^® was obtained using a full-resolution NGI at 39, 60, and 90 L/min, respectively. Two reduced NGI (rNGI) configurations, the filter-only configuration (rNGI-f) and the modified-cup configuration (rNGI-mc), and the fast-screening impactor (FSI) with appropriate inserts to provide a 5-μm cut size were evaluated. The fine particle dose (FPD) obtained using the FSI for Spiriva was statistically similar to that obtained using the full NGI. However, the FPD for both Foradil and Relenza obtained using the FSI was significantly different from that obtained using the full NGI. Despite this, no significant differences were observed for the fine particle fraction (FPF) obtained using the FSI relative to that obtained from the full NGI for any of the DPIs. The use of abbreviated impactor systems appears promising with good agreement observed with the full-resolution NGI, except for small differences observed for the rNGI-mc configuration. These small differences may be product- and/or flow rate-specific, and further evaluation will be required to resolve these differences.     

Depth profiling of the elemental surface composition of the oral microorganismS. salivarius HB and fibrillar mutants by X-ray photoelectron spectroscopy

X-ray photoelectron spectroscopy (XPS) on microbial cell surfaces requires freeze-drying of cells, and as a result, the cell surface appendages flatten out on the cell surface and form a collapsed fibrillar mass. At present, it is unclear how the density, length and composition of these fibrils influence the elemental surface composition as probed by XPS. The sampling depth of XPS can be varied by changing the electron take-off angle. In this article, we made a depth profiling of the collapsed fibrillar mass ofStreptococcus salivarius HB and fibril-deficient mutants by angle-dependent XPS. Methylamine tungstate negative staining and ruthenium red staining followed by sectioning revealed distinct classes of fibrils with various lengths on each of the strains. Interpretation of the angle dependence of the oxygen/carbon (O/C) and phosphorus/carbon (P/C) surface concentration ratios of these strains was difficult. However, the angle dependence of the nitrogen/carbon (N/C) surface concentration ratio could be fully interpreted: N/C did not vary with sampling depth on a bald strain,S. salivarius HBC12 and onS. salivarius HB7, a strain with a dense array of fibrils of uniform length. N/C decreased with sampling depth in case of a sparsely fibrillated strain,S. salivarius HBV51 and eventually reached the value observed for the bald strain, HBC12. A high N/C at small sampling depth was observed forS. salivarius HB with protruding, protein rich fibrils. We conclude that elemental depth profiling of microbial cell surfaces by XPS can be interpreted to coincide with structural and biochemical information on the cell surface as obtained by electron microscopy and can therefore be considered as a useful technique to study structural features of cell surfaces in combination with electron microscopy.     

Long-run monitoring of bacteria,yeasts and other micromycetes in the air of an industrial conurbation

Three different methods were used for the monitoring of airborne microorganisms: (1). Cultivation of microbes trapped in a single-stage biological impactor directly on a solid agar nutrient medium (meat-pepton agar, Sabouraud's agar, blood agar) in Petri dishes. The repeated yearly course of concentrations of cultivable organisms, or colony-forming units (CFU), was obtained by long-run measurements. (2) Aeresol was trapped by impact on membrane filters, and the microorganisms were cultivated by placing the filters on the agar media as above. (3) Direct microorganism counting in a fluorescence microscope; air was sampled in a four-stage impactor where the aerosol was trapped on microscope slides, and the microorganisms were subsequently stained with fluorescent dyes (fluorescein diacctate, 4;6-diamidino-2-phenylindole and, particular, ethidium bromide).

The highest microorganism counts were obtained by using the fluorescence method, the direct cultivation method gave counts an order of magnitude lower, and the method of cultivation on filters gave values approximately 10 times lower than the conventional cultivation.

High variations in the airborne CFU concentrations over the year were observed in Prague. Over the winter season the variations in the amounts of airborne bacteria and other micromycetes as well as the amounts themselves were lower than in the remaining seasons. In the spring and in the summer, the concentrations of yeasts and other micromycetes were highest, whereas in the autumn the concentrations of the microorganisms decreased. Among the bacteria cultivated form the airborne aerosol, the genera Micrococcus, Bacillus, Neisseria and Corynebacterium predominated. The prevailing genera of micromycetes were Penicillium, Aspergillus and Cladosporium.

The concentrations of microorganisms in free air were also affected by the local weather conditions, temperature in particular, the overall air pollution by aerosols was of minor importance in this respect.     

Influence of meteorological parameters and PM2.5 on the level of culturable airborne bacteria and fungi in Abadan,Iran

In recent years, monitoring of airborne bacteria and fungi concentrations has obtained increasing universal attraction not only for influences on ecological balance but also for evaluating their public health consequences. In this study, we aimed to investigate culturable airborne bacteria and fungi levels in different sites of Abadan, and their association with meteorological parameters and PM_2.5 levels. Abadan is one of the most industrialized cities in the southwest of Iran where over the current decade has experienced lots of dust storm episodes. In total, 400 air samples were collected in 6 months (autumn and winter) using a single-stage viable Andersen cascade impactor for sampling airborne bacteria and fungi and portable DustTrak Aerosol Monitor 8520 for measuring PM_2.5 concentrations and meteorological parameters. Microbial concentrations showed a significant difference between various sites over the study period with averages of 569.57?±?312.64 and 482.73?±?242.86 CFU/M³ for bacteria and fungi, respectively. The air temperature had a significant effect on the concentration of both airborne bacteria and fungi. A significant positive correlation between relative humidity and fungi but no correlation between relative humidity and bacteria concentrations were observed. The average airborne PM_2.5 concentrations of all sites among the study period was 93.24?±?116.72 μg/m³. The atmospheric bacterial and fungal communities were strongly positively correlated with the ambient PM_2.5 level. The levels of airborne bacteria and fungi along with PM_2.5 in the air of the city were relatively higher than the recommended levels. Therefore, the best course of action is needed to control emission sources. Further studies are also needed to evaluate the clinical analysis of the health effects of exposure to these pollutants.

     

Evaluation of an Abbreviated Impactor for Fine Particle Fraction (FPF) Determination of Metered Dose Inhalers (MDI)

Abbreviated impactors have been developed recently to allow more rapid evaluation of inhalation products as alternates to the eight-stage Andersen Cascade Impactor (ACI) which has been widely used in the pharmaceutical industry for assessing aerodynamic particle size distribution. In this paper, a two-stage abbreviated impactor, Westech Fine Particle Dose Impactor (WFPD), was used to characterize the aerodynamic particle size of metered dose inhaler (MDI) products, and the results were compared with those obtained using the standard eight-stage ACI. Seven commercial MDI products, with different propellants (chlorofluorocarbon/hydrofluoroalkane) and formulation types (suspension/solution, dry/normal/wet), were tested in this study by both WFPD and ACI. Substantially equivalent measures of fine particle fraction were obtained for most of the tested MDI products, but larger coarse particle fraction and extra-fine particle fraction values were measured from WFPD relative to those measured using the ACI. Use of the WFPD also produced more wall loss than the ACI. Therefore, it is recommended that the system suitability be evaluated on a product-by-product basis to establish substantial equivalency before implementing an abbreviated impactor measurement methodology for routine use in inhaler product characterization.     

Levels of airborne biological agents and related factors in indoor environments of fish toxicity laboratory

In this study, we evaluated the levels of airborne biological agents, such as bacteria, fungi, endotoxin, and (1→3)-β-d-glucan in university fish toxicity laboratory every month for one year and assessed the associated environmental factors. A single-stage viable cascade impactor connected with a pump was used for culturable bacteria and fungi. An analysis of airborne endotoxin and (1→3)-β-d-glucan was performed using kinetic Limulus amebocyte lysate assays. Levels of culturable bacteria and fungi were the highest in summer, whereas levels of endotoxin and (1→3)-β-d-glucan were the highest in winter and spring, respectively. Human activity was correlated with culturable bacteria and fungi, and culturable fungi were also associated with culturable bacteria. Although additional studies based on advanced analyzing technology are required, simultaneous sampling with biomarkers of bacteria is required to further elucidate the characteristics of biological agents.     

Molecular surface characterization of oral streptococci by Fourier transform infrared spectroscopy

In order to characterize the molecular composition of oral streptococci, infrared transmission spectroscopy on freeze-dried cells dissolved in KBr was used. All infrared spectra show similar absorption bands for the strains studied with the most important absorption bands located at 2930 cm-1 (CH), 1653 cm-1 (AmI), 1541 cm-1 (AmII) and two bands at 1236 cm-1 and 1082 cm-1, which were assigned to phosphate and sugar groups. However, calculation of absorption band ratios normalized with respect to the integrated intensity of the CH stretching region around 2930 cm-1, show significant differences between the strains. Both Streptococcus mitis strains possess high AmI/CH and AmII/CH absorption band ratios compared to the other strains. Streptococcus salivarius HBC12, a mutant strain devoid of all proteinaceous surface appendages, shows significantly lower AmI/CH and AmII/CH band ratios with respect to its parent strain S. salivarius HB. Two positive relationships could be established both between the AmII/CH absorption band ratio and the N/C elemental surface concentration ratio of the strains previously, determined from X-ray photoelectron spectroscopy (XPS) and also between AmI/CH and the fraction of carbon atoms at the surface involved in amide bonds, determined by XPS as well. From this comparison, it is concluded that transmission infrared spectroscopy can be employed as a technique to study the molecular surface composition of freeze-dried microorganisms.     

Isolation and Identification of Photosynthetic Bacteria (Rhodospirillaceae) from Antarctic Marine and Freshwater Sediments

Sediment samples obtained from three freshwater lakes and off-shore coastal marine waters on Signy Island, South Orkney Islands, Antarctica have been inoculated into selective enrichment media for purple non-sulphur bacteria (Rhodospirillaceae). From the freshwater sediments strains of Rhodopseudomonas sphaeroides (1), Rhodopseudomonas palustris (1), Rhodospirillum fulvum (1), Rhodospirillum molischanum (1) and Rhodomicrobium vannielii (3) have been isolated. The only purple non-sulphur bacteria obtained from Lake 10 (Amos lake) were strains of Rhodomicrobium vannielii which were able to tolerate hydrogen sulphide (up to 0.04% w/v) found in this lake. Growth of all the other isolates is inhibited by the presence of hydrogen sulphide. Marine sediments yielded strains of Rhodopseudomonas palustris and Rhodomicrobium vannielii . All the isolates grow optimally at temperatures between 25 and 30 °C. mean generation times vary between 8 and 10.7 h depending on species. There is no evidence of cold adaptation in any of the strains studied.     

Fungal and actinomycete spore aerosols measured at different humidities with an aerodynamic particle sizer.

An aerodynamic particle sizer (APS) that uses laser Doppler velocimetry was used to determine aerodynamic diameters of spores of fungal and thermophilic actinomycete species common in mouldy hay, aerosolized at different humidities and temperatures. Results were compared with those obtained from inertial impaction in a cascade impactor. The APS gave slightly smaller measurements than the cascade impactor. Both methods gave aerodynamic diameters generally slightly smaller than the average spore dimensions observed on cascade impactor slides with a microscope. The latter measurements were less than axial dimensions given in the literature. Brief passage of spores through air at 95% relative humidity (RH) and 38 degrees C, compared with 40% RH and 20 degrees C, caused an immediate increase in their aerodynamic diameter and the breaking of chains of spores. Cultures maintained at 75% RH and aerosolized at 98% RH similarly produced larger spore particles than those passed through dry air. These findings have implications for mould-induced asthma and allergic alveolitis since they relate to physical behaviour of airborne spores and particle deposition sites in the lung.     

Influence of meteorological factors on the level and characteristics of culturable bacteria in the air in Gliwice,Upper Silesia (Poland)

Numerous studies have focused on occupational and indoor environments because people spend more than 90% of their time in them. Nevertheless, air is the main source of bacteria in indoors, and outdoor exposure is also crucial. Worldwide studies have indicated that bacterial concentrations vary among different types of outdoor environments, with considerable seasonal variations as well. Conducting comprehensive monitoring of atmospheric aerosol concentrations is very important not only for environmental management but also for the assessment of the health impacts of air pollution. To our knowledge, this is the first study to present outdoor and seasonal changes of bioaerosol data regarding an urban area of Poland. This study aimed to characterize culturable bacteria populations present in outdoor air in Gliwice, Upper Silesia Region, Poland, over the course of four seasons (spring, summer, autumn and winter) through quantification and identification procedures. In this study, the samples of bioaerosol were collected using a six-stage Andersen cascade impactor (with aerodynamic cut-off diameters of 7.0, 4.7, 3.3, 2.1, 1.1 and 0.65 μm). Results showed that the concentration of airborne bacteria ranged from 4 CFU m^?3, measured on one winter day, to a maximum equal to 669 CFU m^?3 on a spring day. The average size of culturable bacterial aerosol over the study period was 199 CFU m^?3. The maximal seasonally averaged concentration was found in the spring season and reached 306 CFU m^?3, and the minimal seasonally averaged concentration was found in the winter 49 CFU m^?3. The most prevalent bacteria found outdoors were gram-positive rods that form endospores. Statistically, the most important meteorological factors related to the viability of airborne bacteria were temperature and UV radiation. These results may contribute to the promotion and implementation of preventative public health programmes and the formulation of recommendations aimed at providing healthier outdoor environments.     

Relative Precision of Inhaler Aerodynamic Particle Size Distribution (APSD) Metrics by Full Resolution and Abbreviated Andersen Cascade Impactors (ACIs): Part 2—Investigation of Bias in Extra-Fine Mass Fraction with AIM-HRT Impactor

The purpose of this study was to resolve an anomalously high measure of extra-fine particle fraction (EPF) determined by the abbreviated cascade impactor possibly relevant for human respiratory tract (AIM-HRT) in the experiment described in Part 1 of this two-part series, in which the relative precision of abbreviated impactors was evaluated in comparison with a full resolution Andersen eight-stage cascade impactor (ACI). Evidence that the surface coating used to mitigate particle bounce was laterally displaced by the flow emerging from the jets of the lower stage was apparent upon microscopic examination of the associated collection plate of the AIM-HRT impactor whose cut point size defines EPF. A filter soaked in surfactant was floated on top of this collection plate, and further measurements were made using the same pressurized metered-dose inhaler-based formulation and following the same procedure as in Part 1. Measures of EPF, fine particle, and coarse particle fractions were comparable with those obtained with the ACI, indicating that the cause of the bias had been identified and removed. When working with abbreviated impactors, this precaution is advised whenever there is evidence that surface coating displacement has occurred, a task that can be readily accomplished by microscopic inspection of all collection plates after allowing the impactor to sample ambient air for a few minutes.     

A case study of airborne culturable microorganisms in a poultry slaughterhouse in Styria, Austria

According to Council Directive 90/679/EEC on the protection of workers from risks related to exposure to biological agents at work, nature, degree and duration of workers’ exposure to microorganisms must be determined. This directive has already been implemented in waste and wastewater management. The present case study investigates concentration and composition of microorganisms in a poultry slaughterhouse in the State of Styria, Austria. From June to November 2002, measurements were conducted at the sampling sites ‘moving rail’ and ‘gall bladder separation’ using the Andersen six stage viable cascade impactor and the SKC BioSampler. The results of this study were compared with other previous studies which were carried out using the same device (ACFM) and the same measurement methods. At the processing area of the ‘moving rail’, the median concentration of airborne mesophilic bacteria was 1.7×10⁶ CFU/m³ which is 8000 times higher than the background concentration of residential areas (approx. 210 CFU/m³). The airborne microorganisms concentration was 1.7×10⁴ CFU/m³ at composting plants which is 100 times lower than at a workplace of a poultry slaughterhouse. The study shows that poultry slaughterhouse employees are exposed to high concentrations of airborne microorganisms throughout the entire work time without using a respiratory protective device. For the protection of employees against airborne biological agents, relevant measures should be introduced to this field of work.     

Fungal and actinomycete spore aerosols measured at different humidities with an aerodynamic particle sizer

T.M. MADELIN AND H.E. JOHNSON. 1992. An aerodynamic particle sizer (APS) that uses laser Doppler velocimetry was used to determine aerodynamic diameters of spores of fungal and thermophilic actinomycete species common in mouldy hay, acrosolized at different humidities and temperatures. Results were compared with those obtained from inertial impaction in a cascade impactor. The APS gave slightly smaller measurements than the cascade impactor. Both methods gave aerodynamic diameters generally slightly smaller than the average spore dimensions observed on cascade impactor slides with a microscope. The latter measurements were less than axial dimensions given in the literature. Brief passage of spores through air at 95% relative humidity (RH) and 38°C, compared with 40% RH and 20°C, caused an immediate increase in their aerodynamic diameter and the breaking of chains of spores. Cultures maintained at 75% RH and aerosolized at 98% RH similarly produced larger spore particles than those passed through dry air. These findings have implications for mould-induced asthma and allergic alveolitis since they relate to physical behaviour of airborne spores and particle deposition sites in the lung.     

Identification of ice-nucleating active Pseudomonas fluorescens strains for biological control of overwintering Colorado potato beetles (Coleoptera: Chrysomelidae)

Laboratory studies were conducted to identify ice-nucleating active bacterial strains able to elevate the supercooling point, the temperature at which freezing is initiated in body fluids, of Colorado potato beetles, Leptinotarsa decemlineata (Say), and to persist in their gut. Adult beetles fed ice-nucleating active strains of Pseudomonas fluorescens, P. putida, or P. syringae at 10(6) or 10(3) bacterial cells per beetle had significantly elevated supercooling points, from -4.5 to -5.7 degrees C and from -5.2 to -6.6 degrees C, respectively, immediately after ingestion. In contrast, mean supercooling point of untreated control beetles was -9.2 degrees C. When sampled at 2 and 12 wk after ingestion, only beetles fed P. fluorescens F26-4C and 88-335 still had significantly elevated supercooling points, indicating that these strains of bacteria were retained. Furthermore, beetle supercooling points were comparable to those observed immediately after ingestion, suggesting that beetle gut conditions were favorable not only for colonization but also for expression of ice-nucleating activity by these two strains. The results obtained from exposure to a single, low dose of either bacterial strain also show that a minimum amount of inoculum is sufficient for establishment of the bacterium in the gut. Persistence of these bacteria in Colorado potato beetles long after ingestion was also confirmed using a polymerase chain reaction technique that detected ice-nucleating active bacteria by virtue of their ina genes. Application of these ice-nucleating active bacteria to elevate the supercooling point of this freeze-intolerant insect pest could significantly reduce their winter survival, thereby reducing local populations and, consequently, crop damage.     

Monitoring airborne genotoxicants in the rubber industry using genotoxicity tests and chemical analyses

This research was designed to examine the presence of mutagenic/carcinogenic compounds in airborne pollutants in the rubber industry using an integrated chemical/biological approach. Inhalable airborne particulate matter (PM-10: <10 microm) was collected in four rubber factories using a high-volume sampler equipped with a cascade impactor for particle fractionation. The organic extracts of two different fractions (0.5-10 microm and <0.5 microm) were examined for mutagenicity with the Ames test and for in vitro DNA-damaging activity in human leukocytes by single-cell microgel electrophoresis (Comet assay). The extracts were also studied by gas chromatography/mass spectrometry (GC/MS) for polycyclic aromatic hydrocarbon (PAH) content. Nitrosamines in ambient air were sampled on cartridges and analysed by GC with a thermal energy analyser (TEA) detector. Airborne volatile genotoxins were monitored in situ using a clastogenicity plant test (Tradescantia/micronuclei test). The results showed that airborne particulates were mainly very fine (<0.5 microm) and that trace amounts of genotoxic nitrosamines (N-nitrosodimethylamine: 0.10-0.98 microg/m(3); N-nitrosomorpholine: 0.77-2.40 microg/m(3)) and PAH (total PAH: 0.34-11.35 microg/m(3)) were present in air samples. Some extracts, particularly those obtained from the finest fractions, were mutagenic with the Ames test and genotoxic with the Comet assay. In situ monitoring of volatile mutagens using the Tradescantia/micronuclei test gave positive results in two working environments. The results showed the applicability of this integrated chemical-biological approach for detecting volatile and non-volatile genotoxins and for monitoring genotoxic hazards in the rubber industry.     

Conformational study and hydrogen bonds detection on elastin-related polypeptides using X-ray photoelectron spectroscopy

The chemical bonds of the pentapeptide sequence of elastin ValGlyGlyValGly (VGGVG), both in its monomer and polymer forms, were correlated with their XPS spectra through a well-established curve-fitting procedure. To aid in this correlation, the C1s, O1s, and N1s chemical shifts of the Boc-VGGVG-OEt, were validated by theoretical calculations, performed in the framework of the Koopman approximation of HF/6-31G molecular orbitals, leading to the "preferred" conformation of the protected monomer. Then the same curve-fitting procedure was adopted for interpreting the XPS spectra of the polypentapeptide as a powder, and the XPS results obtained both for monomer and polymer compounds were compared with those obtained by FT-IR. The polymer was then analyzed after deposition onto a silicon substrate, Si(100), either from methanol or water suspensions and the presence of hydrogen bonds was detected at the polymer/substrate interface and between the polymer chains. The "surface rearrangement" that could be inferred from XPS results strongly confirms that derived from AFM images previously obtained under the same experimental conditions. In particular, the observed amyloid conformation is stabilized by hydrogen bonds to water molecules included in the structure while the formation of the beaded string structure observed in deposits from methanolic suspension is probably mediated by hydrogen bonds to the hydrated silicon surface.     

Measuring the spermosphere colonizing capacity (spermosphere competence) of bacterial inoculants

Spermosphere establishment by bacteria which were coated onto seeds was studied using soybean seeds treated with four bacterial strains at levels of log10 1 to 4 colony-forming units (cfu) per seed planted in a field soil mix, and incubated 48 h. Each strain at every inoculum level developed spermosphere population densities of log10 4 to 8 cfu/seed, demonstrating an average multiplication of log10 3 cfu/seed. An alternative method was developed to differentially rank bacteria for spermosphere colonizing capacity, based upon incorporation of bacteria into a soil and monitoring the resulting spermosphere population densities around noninoculated seeds after 4 days at 14 degrees C. Fifty-seven bacterial strains which were isolated from soybean roots or from water samples, including Pseudomonas putida, P. putida biovar B, P. fluorescens, Serratia liquefaciens, Enterobacter aerogenes, and Bacillus spp. were tested in the spermosphere colonization assay. Average spermosphere population densities for the 57 strains ranged from 0 to log10 7.0 cfu/seed. Strains of a given taxon demonstrated marked diversity with ranges from 0 to log10 6.0 cfu/seed for Bacillus spp. and from log10 1.4 to 7.0 cfu/seed for Pseudomonas putida. The relative ranking of representative strains was consistent in repeating experiments. The potential usefulness of the assay for efforts to develop competitive bacterial inoculants for crop seeds is discussed.     

Lysogeny of Oenococcus oeni (syn. Leuconostoc oenos) and Study of Their Induced Bacteriophages

A large number of strains of Oenococcus oeni (formerly Leuconostoc oenos) that had been isolated from wines were checked for lysogeny with mitomycin C as inducer. As a result of this test, 45% of the strains proved to be lysogenic, suggesting that lysogeny is widespread among bacteria isolated from wines during malolactic fermentation. The sensitivity of bacteria to phages was very different, depending on the strain. All the lysogenic strains were resistant to infection by the temperate phage they released. Some phages infected none of the strains. Phages of Oenoc. oeni had a classical morphology, an isometric head, and a long striated tail. With the broadest host strain as an indicator, phages were detected in wines after malolactic fermentation. Received: 28 November 1997 / Accepted: 5 January 1998