Frequency of F116-mediated transduction of Pseudomonas aeruginosa in a freshwater environment.

Transduction of Pseudomonas aeruginosa streptomycin resistance by a generalized transducing phage, F116, was shown to occur during a 10-day incubation in a flow-through environmental test chamber suspended in a freshwater reservoir. Mean F116 transduction frequencies ranged from 1.4 X 10(-5) to 8.3 X 10(-2) transductants per recipient during the in situ incubation. These transduction frequencies were comparable to transduction frequencies determined in preliminary laboratory transduction experiments. The results demonstrate the potential for naturally occurring transduction in aquatic environments and concurrent environmental and ecological ramifications.     

Frequency of F116-mediated transduction of Pseudomonas aeruginosa in a freshwater environment.

Transduction of Pseudomonas aeruginosa streptomycin resistance by a generalized transducing phage, F116, was shown to occur during a 10-day incubation in a flow-through environmental test chamber suspended in a freshwater reservoir. Mean F116 transduction frequencies ranged from 1.4 X 10(-5) to 8.3 X 10(-2) transductants per recipient during the in situ incubation. These transduction frequencies were comparable to transduction frequencies determined in preliminary laboratory transduction experiments. The results demonstrate the potential for naturally occurring transduction in aquatic environments and concurrent environmental and ecological ramifications.     

Transduction of Various R Factors by Phage P1 in Escherichia coli and by Phage P22 in Salmonella typhimurium

R factors fi(+) and fi(-), with various combinations of drug-resistance markers and isolated from independent sources, were transduced by phage P1kc in Escherichia coli and by phage P22 in Salmonella typhimurium. Usually the entire R factor was transduced by P1kc in E. coli, as indicated by the absence of segregation of the drug-resistance markers from their conjugal transferability. In contrast, the patterns of segregation of the drug-resistance markers and their conjugal transferability differed considerably among various R factors after transduction by P22 in S. typhimurium. Transduction frequencies varied among R factors in both transduction systems.     

Seasonal variation of dissolved aluminum concentration in harmonic-type Lake Biwa, Japan

Monthly observations performed on a typical harmonic-type lake, Lake Biwa (latitude 35°15′ N, longitude 136°05′ E, Japan), showed that the particulate aluminum concentration varied around a mean value of 0.8 μM, with occasional extraordinarily high values, whereas the dissolved aluminum concentration varied, in the surface layer of the whole lake, between a minimum of 0.01 μM and a maximum of 0.30 μM, depending on the season. Although the variation in dissolved aluminum paralleled the variation in the pH of the lake water, the variation in dissolved aluminum lagged behind the variation in pH by approximately 1 month. A series of laboratory incubation experiments suggested that the supply of dissolved aluminum from, and its removal by, the suspended particulate matter involved a slow, pH-dependent reaction. The stoichiometry and the apparent equilibrium constant of this reaction were evaluated by adopting a zeolite-like structure for the surfaces of the suspended particulates. Received: December 28, 2000 / Accepted: August 22, 2001     

Transformation and Transduction in Recombination-defective Mutants of Bacillus subtilis

The effects on transformation and transduction of an ultraviolet sensitivity (uvr(-)) and two ultraviolet sensitivity-recombination deficiency (rec-1(-) and rec-2(-)) mutations in isogenic strains of Bacillus subtilis were investigated. Transformation frequency in the rec-1(-) and rec-2(-) strains was reduced to approximately 5 and 25%, respectively, of the parental strains. Normal kinetics of deoxyribonucleic acid dose response in transformation were found for the rec-1(+) and rec-2(-) strains. Biphasic curves were obtained with the rec-1(-) strains. Transduction frequency with bacteriophage SP-10 decreased parallel to transformation frequency in the rec-1(-) and rec-2(-) strains. This result suggests that transformation and SP-10 transduction share a common mechanism for genetic recombination. It also indicates that the reduction in transformation frequency of these strains was not due to altered competence. Transduction frequency with bacteriophage PBS-1 or 3NT, on the contrary, was not diminished in rec-1(-) strains. This frequency was reduced in rec-2(-) strains but not as severely as that of transformation or SP-10 transduction. Several hypotheses to interpret these differences are presented. Recombination frequency between linked markers was reduced more than 50% in transformation by the presence of the rec-1(-) mutation. Linkage was unaffected in the rec-2(-) strains. Neither the rec-1(-) nor the rec-2(-) mutation had an effect on linkage in PBS-1 or 3NT transduction. The uvr(-) strains were transformed at a frequency equal to or greater than that of the parental strains. These strains were transduced by all bacteriophage systems at frequencies about twofold higher than those of parental strains.     

Climate‐related changes of soil characteristics affect bacterial community composition and function of high altitude and latitude lakes

Lakes at high altitude and latitude are typically unproductive ecosystems where external factors outweigh the relative importance of in‐lake processes, making them ideal sentinels of climate change. Climate change is inducing upward vegetation shifts at high altitude and latitude regions that translate into changes in the pools of soil organic matter. Upon mobilization, this allochthonous organic matter may rapidly alter the composition and function of lake bacterial communities. Here, we experimentally simulate this potential climate‐change effect by exposing bacterioplankton of two lakes located above the treeline, one in the Alps and one in the subarctic region, to soil organic matter from below and above the treeline. Changes in bacterial community composition, diversity and function were followed for 72 h. In the subarctic lake, soil organic matter from below the treeline reduced bulk and taxon‐specific phosphorus uptake, indicating that bacterial phosphorus limitation was alleviated compared to organic matter from above the treeline. These effects were less pronounced in the alpine lake, suggesting that soil properties (phosphorus and dissolved organic carbon availability) and water temperature further shaped the magnitude of response. The rapid bacterial succession observed in both lakes indicates that certain taxa directly benefited from soil sources. Accordingly, the substrate uptake profiles of initially rare bacteria (copiotrophs) indicated that they are one of the main actors cycling soil‐derived carbon and phosphorus. Our work suggests that climate‐induced changes in soil characteristics affect bacterioplankton community structure and function, and in turn, the cycling of carbon and phosphorus in high altitude and latitude aquatic ecosystems.     

Differential response of high-elevation planktonic bacterial community structure and metabolism to experimental nutrient enrichment

Nutrient enrichment of high-elevation freshwater ecosystems by atmospheric deposition is increasing worldwide, and bacteria are a key conduit for the metabolism of organic matter in these oligotrophic environments. We conducted two distinct in situ microcosm experiments in a high-elevation lake (Emerald Lake, Sierra Nevada, California, USA) to evaluate responses in bacterioplankton growth, carbon utilization, and community structure to short-term enrichment by nitrate and phosphate. The first experiment, conducted just following ice-off, employed dark dilution culture to directly assess the impact of nutrients on bacterioplankton growth and consumption of terrigenous dissolved organic matter during snowmelt. The second experiment, conducted in transparent microcosms during autumn overturn, examined how bacterioplankton in unmanipulated microbial communities responded to nutrients concomitant with increasing phytoplankton-derived organic matter. In both experiments, phosphate enrichment (but not nitrate) caused significant increases in bacterioplankton growth, changed particulate organic stoichiometry, and induced shifts in bacterial community composition, including consistent declines in the relative abundance of Actinobacteria. The dark dilution culture showed a significant increase in dissolved organic carbon removal in response to phosphate enrichment. In transparent microcosms nutrient enrichment had no effect on concentrations of chlorophyll, carbon, or the fluorescence characteristics of dissolved organic matter, suggesting that bacterioplankton responses were independent of phytoplankton responses. These results demonstrate that bacterioplankton communities in unproductive high-elevation habitats can rapidly alter their taxonomic composition and metabolism in response to short-term phosphate enrichment. Our results reinforce the key role that phosphorus plays in oligotrophic lake ecosystems, clarify the nature of bacterioplankton nutrient limitation, and emphasize that evaluation of eutrophication in these habitats should incorporate heterotrophic microbial communities and processes.     

Formation,Transport and Retention of Aggregates in a River‐Lake System (Spree,Germany)

The formation, particle size distribution, structure and retention of aggregates were studied in a river‐lake system of the lowland River Spree and factors influencing these processes were evaluated. Samples were taken from the inflow and outflow of a flushed lake, as well as along the adjacent flowing reach of the River Spree, between April 2001 and May 2002. The aim was to record the influence of the seasons and different turbulence intensities on the size, shape, composition, abundance and transport properties of the particles. As additional parameters, the phytoplankton biomass, the concentration of suspended particulate matter, the concentrations of nutrients and the discharge were measured. The abundance of the particles showed a strong seasonality in the investigated period, which closely correlated with the concentrations of suspended particulate matter and particulate organic matter in the water column. The organic carbon content of the suspended matter varied between 37% (dry weight) in summer and about 14% in winter. The concentration of particles ranged between 500 and 2500 particles/ml. The number of particles was significantly lower upstream from the lake than at the downstream transects. The majority of the aggregates in the River Spree were smaller than 500 µm, most of the particles even showing a diameter less than 50 µm. Many particles, above all algae, were washed out of the lake Neuendorfer See into the adjacent flowing reach of the River Spree. Algae influenced the abundance of the particles and contributed to the formation of aggregates with their stickiness. In the river flowing section large particles were formed by turbulence structures, whilst simultaneously the number of smaller particles decreased in the water column caused by aggregation processes. In general, an increasing flow velocity and discharge rate resulted in a larger amount of aggregates in the water column. On the other hand, particle abundance was reduced in the flow direction when the flow velocity was low. Under these conditions particles were retained by sedimentation and benthic filter‐feeding. (© 2005 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

The effect of human activities on benthic macroinvertebrate diversity in tributary lagoons surrounding Lake Biwa

In aquatic ecosystems, tributaries play an important role in maintaining the populations and diversity of aquatic organisms throughout the drainage basin, but because of their small catchments, these ecosystems are often more susceptible to the effect of land-use changes and flow-regime alterations. Here, we examined anthropogenic effects on benthic macroinvertebrate diversity in the tributary lagoons, collectively called “Naiko,” surrounding Lake Biwa. We sampled macroinvertebrates and determined the environmental characteristics of 20 tributary lagoons. We identified the environmental factors determining the diversity of macroinvertebrates and found that turbidity significantly diminished species diversity. We assessed the anthropogenic stressors that contributed to the increase in turbidity and found that human population density and the proportion of paddy fields in the watershed area were positively correlated with turbidity, most likely caused by erosion of terrigenous organic matter from the paddy fields and urban areas. In addition, the presence of sluice gates and the lengths of channels connecting to the main lake were positively correlated with turbidity, suggesting that these factors lowered hydrologic connectivity and retained organic matter. We concluded that changes to the landscape and decreased hydrologic connectivity caused by human activity increased turbidity, which in turn decreased macroinvertebrate diversity. The identification of these factors in tributaries is vital for developing a strategy for habitat restoration to conserve the entire ecosystem of the Lake Biwa basin.     

Remote sensing for lake research and monitoring – Recent advances

Lakes are important ecosystems providing various ecosystem services. Stressors such as eutrophication or climate change, however, threaten their ecological functions. National and international legislations address these threats and claim consistent, long-term monitoring schemes. Remote sensing data and products provide synoptic, spatio-temporal views and their integration can lead to a better understanding of lake ecology and water quality. Remote sensing therefore gains increasing awareness for analysing water bodies. Various empirical and semi-analytical algorithms exist to derive remote sensing indicators as proxies for climate change or ecological response variables. Nevertheless, most monitoring networks lack an integration of remote sensing data. This review article therefore provides a comprehensive overview how remote sensing can support lake research and monitoring. We focus on remote sensing indicators of lake properties, i.e. water transparency (suspended particulate matter, coloured dissolved organic matter, Secchi disc depth, diffuse attenuation coefficient, turbidity), biota (phytoplankton, cyanobacteria, submerged and emerged aquatic vegetation), bathymetry, water temperature (surface temperature) and ice phenology (ice cover, ice-on, ice-out). After a brief background introducing principles of lake remote sensing we give a review on available sensors and methods. We categorise case studies on remote sensing indicators with respect to lake properties and processes. We discuss existing challenges and benefits of integrating remote sensing into lake monitoring and ecological research including data availability, ready-to-use tools and accuracies.     

Potential for transduction of plasmids in a natural freshwater environment: effect of plasmid donor concentration and a natural microbial community on transduction in Pseudomonas aeruginosa

Transduction of Pseudomonas aeruginosa plasmid Rms149 by the generalized transducing bacteriophage phi DS1 was shown to occur during a 9-day incubation of environmental test chambers in a freshwater reservoir. Plasmid DNA was transferred from a nonlysogenic plasmid donor to a phi DS1 lysogen of P. aeruginosa that served both as the source of the transducing phage and as the recipient of the plasmid DNA. When the concentration of donors introduced into the chambers was varied while the recipient concentration in each chamber was at a level equivalent to natural concentrations of P. aeruginosa, the concentration of plasmid-containing donor cells introduced was shown to affect the frequency of transduction significantly. Transduction was observed both in the absence and in the presence of the natural microbial community. The presence of the natural community resulted in a rapid decrease in the numbers of the introduced donors and recipients and a decrease in the number of transductants recovered. These results demonstrate the potential for naturally occurring transduction in aquatic environments and indicate that donor load may be an important parameter in assessing this potential.     

Protein Particulate Detection Issues in Biotherapeutics Development—Current Status

Formation of aggregates and particulates in biopharmaceutical formulation continues to be one of the major quality concerns in biotherapeutics development. The presence of large quantities of aggregates is believed to be one of the causes of unwanted immunogenic responses. Protein particulates can form in a wide range of sizes and shapes. Therefore, a comprehensive characterization of particulates in biologics formulation continues to be challenging. The quantity of small size aggregates (e.g., dimer) in a stable biologics formulation is well controlled using precision analytical techniques (e.g., high-performance liquid chromatography). Particulate in clinical and commercial formulations is monitored using visual inspection and subvisible particulate counting assays. While visual inspection (by human eye or automated systems) is intended to detect particulates (intrinsic and extrinsic) of ~100 μm or larger, the subvisible counting methods cover smaller size ranges down to 10 μm. It is well recognized that research of particulates in the submicron (<1 μm) and low-micron (1–10 μm) ranges may provide important clues to understand the mechanism of particulate formation. The recent years have seen a significant increase in the development of newer technologies for more comprehensive characterization of particulates. This is attributed to increased awareness in this field of research over the past 5 years, stimulated by scholarly articles, commentaries, and robust discussions in various forums. This article provides an overview of emerging detection technologies that provide complementary characterization data encompassing a wider size range of particulates. It also discusses their advantages and limitations in the context of applications in biotherapeutics development.KEY WORDS: biotherapeutics, formulation development, laser diffraction, particulate matter, protein aggregation     

Mutagenicity studies on complex environmental mixtures: selection of solvent system for extraction.

The mutagenic activities in the Salmonella/microsome assay of dichloromethane (DCM) and acetone extracts of complex environmental mixtures were compared. The particulate samples used in the IPCS collaborative study were Soxhlet-extracted twice with DCM followed by a third extraction with acetone. Compared with the mutagenic activity of the first extract, the third (acetone) extract of the urban particulate matter showed a relatively high mutagenic activity. In contrast to this the third extract of the diesel particulate matter contributed very little additional mutagenic activity. Furthermore, 10 filter samples of air particulates from a suburban airport area were collected for comparison of the extraction efficiency of DCM and acetone. Each sample was divided into two samples of identical size followed by extraction with acetone and DCM, respectively. No clear difference in the mutagenic activity of these extracts was observed in strains TA98 and TA98NR. It is concluded that for ambient air particulates (but not emission samples) acetone may extract some mutagenic compounds which are not extracted by DCM. The amount of these additional extractable compounds seems to depend on the composition of the sample. As DCM extracts are better suited for further fractionation and chemical analysis DCM is considered to be the best choice for a general solvent system for extraction of complex environmental mixtures.     

Potential for transduction of plasmids in a natural freshwater environment: effect of plasmid donor concentration and a natural microbial community on transduction in Pseudomonas aeruginosa.

Transduction of Pseudomonas aeruginosa plasmid Rms149 by the generalized transducing bacteriophage phi DS1 was shown to occur during a 9-day incubation of environmental test chambers in a freshwater reservoir. Plasmid DNA was transferred from a nonlysogenic plasmid donor to a phi DS1 lysogen of P. aeruginosa that served both as the source of the transducing phage and as the recipient of the plasmid DNA. When the concentration of donors introduced into the chambers was varied while the recipient concentration in each chamber was at a level equivalent to natural concentrations of P. aeruginosa, the concentration of plasmid-containing donor cells introduced was shown to affect the frequency of transduction significantly. Transduction was observed both in the absence and in the presence of the natural microbial community. The presence of the natural community resulted in a rapid decrease in the numbers of the introduced donors and recipients and a decrease in the number of transductants recovered. These results demonstrate the potential for naturally occurring transduction in aquatic environments and indicate that donor load may be an important parameter in assessing this potential.     

Meiobenthic response to river-borne benthic particulate matter – a microcosm experiment

1. Benthic communities inhabiting river–lake interfaces are confronted with temporal variation in the import of river‐borne material to the associated littoral zone. The primary objective of this study was to evaluate the exposure to river‐borne benthic particulate matter as a force shaping the meiofauna of the river–lake interface. 2. In a 2 × 2 factorial design, combinations of strongly and weakly river‐influenced communities (near or far from the river mouth) with and without river‐borne benthic particulate matter enrichment were studied in laboratory microcosms to assess the response of the heterotrophic meiobenthos to river‐borne matter. 3. Nematodes dominated both habitat types and were the only meiofaunal taxon that responded to the enrichment. 4. A strong interactive effect between experimental enrichment and location of site was observed. Whereas nematodes from the littoral habitat close to the river mouth responded to the enrichment only marginally, nematodes from the remote site were strongly affected in the presence of river‐borne particulate matter. 5. In the non‐enriched samples of the remote site, the number of nematodes increased greatly over time. Among nematode feeding types, deposit feeders dominated over chewers at the beginning of the experiment but both types were equally abundant at the end. In the presence of river‐borne matter the increase of chewers ceased and a shift in feeding type composition did not occur. 6. It was shown that for river–lake interface meiofaunal assemblages in relative proximity, the same taxon might respond differently if exposed to an external pressure, e.g. river‐borne components. These components may have a stronger influence on taxa with longer life cycles.     

Stoichiometric Constraints on Food-Web Dynamics: A Whole-Lake Experiment on the Canadian Shield

A whole-lake manipulation of food-web structure (introduction of a top predator, northern pike, to a minnow-dominated lake) was performed in a Canadian Shield lake (L110) to examine the stoichiometric consequences of changes in planktonic community structure generated by altered food-web structure. Minnow abundance, zooplankton biomass and community composition, microconsumer abundance, and concentration and carbon–phosphorus (C:P) ratio of suspended particulate matter were monitored in L110 and unmanipulated L240 before (1992) and after (1993–95) pike introduction. Algal biomass in L110 determined from microscopic examination for postmanipulation and premanipulation periods was also compared with dynamics in a suite of unmanipulated reference lakes from long-term monitoring records. Pike were added in spring in 1993 and 1994 in sufficient quantity to raise pike biomass to levels of around 22 kg ha^{− 1} by 1994. Minnow populations in L110 responded dramatically, decreasing to levels 30% (1993), 10% (1994), and less than 1% (1995) of premanipulation values. However, most components lower in the food web did not respond in a manner consistent with predictions of existing food-web theory, such as the idea of cascading trophic interactions (CTI). While Daphnia biomass increased in L110 in the first year following manipulation, consistent with CTI, this effect was temporary and Daphnia collapsed in 1995, the year of lowest minnow abundance. Total zooplankton biomass in both lakes declined during the study period and, contrary to CTI, this decline appeared somewhat stronger in L110 than in L240. Dominant microconsumers (heterotrophic microflagellates) did not differ among years in either lake and did not appear to respond to food-web manipulation. At the bottom of the food web, no changes in bacterial biomass occurred in either lake. However, total concentrations of particulate matter appeared to increase in L110 after manipulation (contrary to expectations based on the theory of CTI) while algal biomass did not change in the manipulated lake relative to reference systems. Finally, particulate C:P increased in both L110 and L240 during the study period. The lack of strong response of Daphnia, the lack of response of the microbial food web, decreases in zooplankton biomass and increases in particulate biomass following reduction of minnow populations after piscivore introduction are at odds with expectations from existing food-web theory, such as the idea of CTI as currently formulated. However, the extremely high C:P ratios in particulate matter at the base of the food webs in these lakes, the coincidence of zooplankton declines and increases in particulate C:P ratios, and the results of small-scale mesocosm food-quality experiments are consistent with a hypothesis of a stoichiometric constraint operating on food-web dynamics in this and similar ecosystems. Received 22 April 1997; accepted 8 July 1997.     

Effects of zooplankton on sedimentation in pelagic ecosystems: Theory and test in two lakes of the Canadian shield

A simple heuristic theory based on conservation of matter and describing the fate of autotrophic production in pelagic ecosystems was developed to assist in conceptualizing how zooplankton affect sedimentation processes in lakes and oceans. The theory predicted that effects of zooplankton on the fraction of autotrophic incorporation that sediments (the export ratio) will be a function of factors related both to zooplankton digestion and egestion and to tendencies of particulate matter to sediment directly prior to mineralization. As a result, effects of zooplankton grazing on the export ratio were predicted to be site-dependent, a function of physical conditions and zooplankton communities characterizing an ecosystem. The theory was tested by monitoring autotrophic production of C, N, and P, sedimentation of C, N, and P, and zooplankton biomass in two lakes of the Canadian shield characterized by contrasting morphometry and food-web structure. In Lake 110, a small elongate lake protected from wind exposure, export ratios of C, N, and P declined strongly with zooplankton biomass. In contrast, in L240, a larger lake with considerably greater wind exposure, export ratios increased with zooplankton biomass. These results were consistent with predictions of our theory that effects of zooplankton on sedimentation processes will depend on the tendency of particulate matter to directly sediment relative to the tendency of egested materials to sediment. However, no significant differences in relationships between export ratios for C, N, and P and zooplankton biomass were found.     

Watershed Deforestation and Down-Estuary Transformations Alter Sources,Transport, and Export of Suspended Particles in Panamanian Mangrove Estuaries

We identified eight Panamanian watersheds in which conversion from wet tropical forest to pastures differed and assessed the effects of degree of deforestation, and down-estuary transformations, on the suspended particulate matter discharged from the watersheds, entering, traversing through mangrove estuaries, and emerging into coastal waters. Deforested watersheds discharged larger concentrations of suspended particulate matter, with lower % C and N, higher mineral content, and heavier isotopic signatures into fresh reaches of estuaries. Down-estuary, sediment entrainment increased non-organic content of particulates, and watershed-derived imprints of deforestation on composition of particulate matter were mostly erased by within-estuary transformations. Isotopic signatures of C, N, and S in particulate matter demonstrated strong land-sea couplings, and indicated that the direction of the coupling was asymmetrical, with terrestrial and estuarine sources delivering particulate materials to coastal waters and sediments. Mangrove estuaries therefore both act as powerful modulators of human activities on land, while also exporting particulate materials to sea.     

Mutagenicity of particulates from the laboratory combustion of plastics

Carcinogenic polycyclic aromatic hydrocarbons (PAHs) and nitropolycyclic aromatic hydrocarbons (nitro-PAHs) have been identified in airborne particulate organic matter extracts. The pollutant sources were generally contributed by motor vehicles and industrial activity. Massive quantities of urban solid wastes, containing plastic materials such as PVC, PET, PS, and PE, burnt in the open air in local garbage dumps are frequently found in developing countries. In this study, the smog particulates from the combustion of these synthetic polymers were produced in a laboratory combustion chamber. The mutagenicity of acetone extracts from the smog particulates was evaluated with Salmonella typhimurium TA98 and TA100 in the presence and absence of S9 mix. Four samples in TA98 exhibited higher mutagenicity than those in TA100. The greatest mutagenicity was observed from the extracts of particulates from combustion of PVC followed by that of PS, PET, and PE. To determine the major mutagenic compounds in these samples, mutagens were partially purified through TLC and their mutagenicity was monitored with TA98. 1-NP and DNPs in the above samples were also determined by HPLC. The amounts of 1-NP and DNPs generally corresponded with their mutagenicity. Higher levels of 1-NP and DNPS from the combustion of PVC, PET, and PS. the combustion of synthetic polymer wastes might be responsible for the presence of high levels of 1-NP and DNPs in Taiwan urban air.