Survival of faecal indicator bacteria in tropical estuarine waters (Bangpakong River,Thailand)

AIMS: To investigate the survival of cultivable bacteria in the tropical Bangpakong estuary (Eastern Thailand) under different salinities and light conditions. METHODS AND RESULTS: Dark and light microcosm experiments using membrane diffusion chambers were carried out under three different experimental conditions, namely (i) low salinity, (ii) progressive mixing with brackish water and (iii) fast mixing with high salinity water spiked with raw urban sewage. Faecal coliforms declined faster than faecal enterococci, as shown by survival T90 values ranging from 82.2 +/- 4.2 to 14.5 +/- 0.8 h and 97.5 +/- 0.4-20.6 +/- 1.2 h, respectively. The survival of freshwater heterotrophic bacteria was higher but variable (121.2 +/- 5.0-30.1 +/- 14.3 h), whereas that of heterotrophic marine bacteria was rather stable (81.5 +/- 4.2-44.6 +/- 2.5 h). CONCLUSIONS: Overall survival was higher in low salinities. Light had a further deleterious effect, since it accelerated the decay of faecal indicators, particularly in high salinities. Faecal enterococci had a higher resistance to environmental conditions compared with faecal coliforms. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is relevant to the understanding of the behaviour of different faecal indicator bacteria and the optimization of sewage treatment plants aimed at the reduction and/or elimination of faecal load discharged into estuarine waters submitted to salinity variations.     

Significance of size and nucleic acid content heterogeneity as measured by flow cytometry in natural planktonic bacteria.

Total bacterial abundances estimated with different epifluorescence microscopy methods (4',6-diamidino-2-phenylindole [DAPI], SYBR Green, and Live/Dead) and with flow cytometry (Syto13) showed good correspondence throughout two microcosm experiments with coastal Mediterranean water. In the Syto13-stained samples we could differentiate bacteria with apparent high DNA (HDNA) content and bacteria with apparent low DNA (LDNA) content. HDNA bacteria, "live" bacteria (determined as such with the Molecular Probes Live/Dead BacLight bacterial viability kit), and nucleoid-containing bacteria (NuCC) comprised similar fractions of the total bacterial community. Similarly, LDNA bacteria and "dead" bacteria (determined with the kit) comprised a similar fraction of the total bacterial community in one of the experiments. The rates of change of each type of bacteria during the microcosm experiments were also positively correlated between methods. In various experiments where predator pressure on bacteria had been reduced, we detected growth of the HDNA bacteria without concomitant growth of the LDNA bacteria, such that the percentage contribution of HDNA bacteria to total bacterial numbers (%HDNA) increased. This indicates that the HDNA bacteria are the dynamic members of the bacterial assemblage. Given how quickly and easily the numbers of HDNA and LDNA bacteria can be obtained, and given the similarity to the numbers of "live" cells and NuCC, the %HDNA is suggested as a reference value for the percentage of actively growing bacteria in marine planktonic environments.     

Do Patterns of Bacterial Diversity along Salinity Gradients Differ from Those Observed for Macroorganisms?

It is widely accepted that biodiversity is lower in more extreme environments. In this study, we sought to determine whether this trend, well documented for macroorganisms, also holds at the microbial level for bacteria. We used denaturing gradient gel electrophoresis (DGGE) with phylum-specific primers to quantify the taxon richness (i.e., the DGGE band numbers) of the bacterioplankton communities of 32 pristine Tibetan lakes that represent a broad salinity range (freshwater to hypersaline). For the lakes investigated, salinity was found to be the environmental variable with the strongest influence on the bacterial community composition. We found that the bacterial taxon richness in freshwater habitats increased with increasing salinity up to a value of 1‰. In saline systems (systems with >1‰ salinity), the expected decrease of taxon richness along a gradient of further increasing salinity was not observed. These patterns were consistently observed for two sets of samples taken in two different years. A comparison of 16S rRNA gene clone libraries revealed that the bacterial community of the lake with the highest salinity was characterized by a higher recent accelerated diversification than the community of a freshwater lake, whereas the phylogenetic diversity in the hypersaline lake was lower than that in the freshwater lake. These results suggest that different evolutionary forces may act on bacterial populations in freshwater and hypersaline lakes on the Tibetan Plateau, potentially resulting in different community structures and diversity patterns.     

Assessment of Salinity-Related Mortality of Freshwater Bacteria in the Saint Lawrence Estuary

The growth response of freshwater bacteria from the St. Lawrence River, exposed to brackish waters (salinity of 0 to 20(permil)) from the upper estuary, was assessed by a methodology requiring the combined use of dilution cultures and diffusion chambers. The longitudinal distribution of bacterial abundance in waters within this salinity range was also examined. Growth of the freshwater bacteria was reduced by 15 and 50% after exposure to salinities of 10 and 20(permil), respectively. At lower salinities, no growth reduction was observed, and at a salinity of 2(permil), growth was even stimulated. Longitudinal distribution data showed that bacterial abundance also peaked at this salinity. In contrast with an earlier hypothesis, this study shows that the decline of bacterial abundance in the low-salinity waters of the estuary is not caused by salinity-related mortality of freshwater bacteria, because the mixing time between fresh and marine (>20(permil)) waters is relatively long (days). However, results suggest that mortality of freshwater bacteria can be an important process in estuaries with shorter mixing times (hours). The combined use of diffusion chambers and dilution cultures proved to be a valuable methodology for assessing growth (or mortality) of bacteria exposed to environmental gradients.     

Significance of Size and Nucleic Acid Content Heterogeneity as Measured by Flow Cytometry in Natural Planktonic Bacteria

Total bacterial abundances estimated with different epifluorescence microscopy methods (4′,6-diamidino-2-phenylindole [DAPI], SYBR Green, and Live/Dead) and with flow cytometry (Syto13) showed good correspondence throughout two microcosm experiments with coastal Mediterranean water. In the Syto13-stained samples we could differentiate bacteria with apparent high DNA (HDNA) content and bacteria with apparent low DNA (LDNA) content. HDNA bacteria, “live” bacteria (determined as such with the Molecular Probes Live/Dead BacLight bacterial viability kit), and nucleoid-containing bacteria (NuCC) comprised similar fractions of the total bacterial community. Similarly, LDNA bacteria and “dead” bacteria (determined with the kit) comprised a similar fraction of the total bacterial community in one of the experiments. The rates of change of each type of bacteria during the microcosm experiments were also positively correlated between methods. In various experiments where predator pressure on bacteria had been reduced, we detected growth of the HDNA bacteria without concomitant growth of the LDNA bacteria, such that the percentage contribution of HDNA bacteria to total bacterial numbers (%HDNA) increased. This indicates that the HDNA bacteria are the dynamic members of the bacterial assemblage. Given how quickly and easily the numbers of HDNA and LDNA bacteria can be obtained, and given the similarity to the numbers of “live” cells and NuCC, the %HDNA is suggested as a reference value for the percentage of actively growing bacteria in marine planktonic environments.     

Bacterial Growth on Allochthonous Carbon in Humic and Nutrient-enriched Lakes: Results from Whole-Lake 13C Addition Experiments

Organic carbon (C) in lakes originates from two distinct sources—primary production from within the lake itself (autochthonous supply) and importation of organic matter from the terrestrial watershed (allochthonous supply). By manipulating the ¹³C of dissolved inorganic C, thereby labeling within-lake primary production, we examined the relative importance of autochthonous and allochthonous C in supporting bacterial production. For 35 days, NaH¹³CO₃ was added daily to two small, forested lakes. One of the lakes (Peter) was fertilized so that primary production exceeded total respiration in the epilimnion. The other lake (Tuesday), in contrast, was low in productivity and had high levels of colored dissolved organic C (DOC). To obtain bacterial C isotopes, bacteria were regrown in situ in particle-free lake water in dialysis tubes. The contribution of allochthonous C to bacterial biomass was calculated by applying a two-member mixing model. In the absence of a direct measurement, the isotopic signature of the autochthonous end-member was estimated indirectly by three different approaches. Although there was excess primary production in Peter Lake, bacterial biomass consisted of 43–46% allochthonous C. In Tuesday Lake more than 75% of bacterial growth was supported by allochthonous C. Although bacteria used autochthonous C preferentially over allochthonous C, DOC from the watershed contributed significantly to bacterial production. In combination with results from similar experiments in different lakes, our findings suggest that the contribution of allochthonous C to bacterial production can be predicted from ratios of chromophoric dissolved organic matter (a surrogate for allochthonous supply) and chlorophyll a (a surrogate for autochthonous supply).     

Bacterioplankton community composition along a salinity gradient of sixteen high-mountain lakes located on the Tibetan Plateau, China

The influence of altitude and salinity on bacterioplankton community composition (BCC) in 16 high-mountain lakes located at altitudes of 2,817 to 5,134 m on the Eastern Qinghai-Xizang (Tibetan) Plateau, China, spanning a salinity gradient from 0.02% (freshwater) to 22.3% (hypersaline), was investigated. Three different methods, fluorescent in situ hybridization, denaturing gradient gel electrophoresis (DGGE) with subsequent band sequencing, and reverse line blot hybridization (RLB) with probes targeting 17 freshwater bacterial groups, were used for analysis of BCC. Furthermore, the salt tolerances of 47 strains affiliated with groups detected in or isolated from the Tibetan habitats were investigated. Altitude was not found to influence BCC significantly within the investigated range. Several groups of typical freshwater bacteria, e.g., the ACK-M1 cluster and the Polynucleobacter group, were detected in habitats located above 4,400 m. Salinity was found to be the dominating environmental factor controlling BCC in the investigated lakes, resulting in only small overlaps in the BCCs of freshwater and hypersaline lakes. The relative abundances of different classes of Proteobacteria showed a sharp succession along the salinity gradient. Both DGGE and RLB demonstrated that a few freshwater bacterial groups, e.g., GKS98 and LD2, appeared over wide salinity ranges. Six freshwater isolates affiliated with the GKS98 cluster grew in ecophysiological experiments at maximum salinities of 0.3% to 0.7% (oligosaline), while this group was detected in habitats with salinities up to 6.7% (hypersaline). This observation indicated ecologically significant differences in ecophysiological adaptations among members of this narrow phylogenetic group and suggested ecological significance of microdiversity.     

Adaptations of microbial communities and dissolved organics to seasonal pressures in a mesotrophic coastal Mediterranean lake

In lake ecosystems, changes in eukaryotic and prokaryotic microbes and the concentration and availability of dissolved organic matter (DOM) produced within or supplied to the system by allochthonous sources are components that characterize complex processes in the microbial loop. We address seasonal changes of microbial communities and DOM in the largest Croatian lake, Vrana. This shallow lake is connected to the Adriatic Sea and is impacted by agricultural activity. Microbial community and DOM structure were driven by several environmental stressors, including drought, seawater intrusion and heavy precipitation events. Bacterial composition of different lifestyles (free-living and particle-associated) differed and only a part of the particle-associated bacteria correlated with microbial eukaryotes. Oscillations of cyanobacterial relative abundance along with chlorophyll a revealed a high primary production season characterized by increased levels of autochthonous DOM that promoted bacterial processes of organic matter degradation. From our results, we infer that in coastal freshwater lakes dependent on precipitation-evaporation balance, prolonged dry season coupled with heavy irrigation impact microbial communities at different trophic levels even if salinity increases only slightly and allochthonous DOM inputs decrease. These pressures, if applied more frequently or at higher concentrations, could have the potential to overturn the trophic state of the lake.     

Effect of salinity gradients and heterotrophic microbial activity on biodegradation of nitrilotriacetic acid in laboratory simulations of the estuarine environment.

The biodegradation of nitrilotriacetic acid (NTA), a synthetic replacement detergent builder, in the estuarine environment was examined by using a laboratory estuarine simulation. Two interdependent microcosms were used; each of five vessels was equilibrated with a saline gradient between 1.30 and 17.17%, with the final vessel subsequently being increased to a maximum salinity of 31.6%. Each microcosm was seeded simultaneously with heterotrophic bacteria from both fresh and saline sources. Viable counts demonstrated the ability of each microcosm to sustain a mixed heterotrophic bacterial community throughout the range of salinities for 183 days after a stabilization period. Isolation studies demonstrated that both systems contained four bacterial species, representatives of the genera Vibrio and Flavobacterium and members of the coryneform group and the family Enterobacteriaceae. Total bacterial numbers and species diversity decreased with increased salinity. NTA was administered at low and high concentrations, one concentration to each microcosm, initially with the least amount of saline. Removal of both concentrations of NTA occurred and was attributed to biodegradation after a period of bacterial acclimatization. Subsequent dosing of NTA to vessels of higher salinity demonstrated that biodegradation was incomplete at observed mean salinities of greater than 9.18% at low influent NTA concentrations and greater than 5.08% at high influent NTA concentrations. Therefore, acclimatization was dose dependent. It was concluded that NTA acclimatization at the higher salinities ceased because of salinity stress-induced failure of NTA catabolism and not the disappearance of a particular bacterial species.     

Effect of salinity gradients and heterotrophic microbial activity on biodegradation of nitrilotriacetic acid in laboratory simulations of the estuarine environment

The biodegradation of nitrilotriacetic acid (NTA), a synthetic replacement detergent builder, in the estuarine environment was examined by using a laboratory estuarine simulation. Two interdependent microcosms were used; each of five vessels was equilibrated with a saline gradient between 1.30 and 17.17%, with the final vessel subsequently being increased to a maximum salinity of 31.6%. Each microcosm was seeded simultaneously with heterotrophic bacteria from both fresh and saline sources. Viable counts demonstrated the ability of each microcosm to sustain a mixed heterotrophic bacterial community throughout the range of salinities for 183 days after a stabilization period. Isolation studies demonstrated that both systems contained four bacterial species, representatives of the genera Vibrio and Flavobacterium and members of the coryneform group and the family Enterobacteriaceae. Total bacterial numbers and species diversity decreased with increased salinity. NTA was administered at low and high concentrations, one concentration to each microcosm, initially with the least amount of saline. Removal of both concentrations of NTA occurred and was attributed to biodegradation after a period of bacterial acclimatization. Subsequent dosing of NTA to vessels of higher salinity demonstrated that biodegradation was incomplete at observed mean salinities of greater than 9.18% at low influent NTA concentrations and greater than 5.08% at high influent NTA concentrations. Therefore, acclimatization was dose dependent. It was concluded that NTA acclimatization at the higher salinities ceased because of salinity stress-induced failure of NTA catabolism and not the disappearance of a particular bacterial species.     

Predatory Bacteriovorax communities ordered by various prey species

The role of predation in altering microbial communities has been studied for decades but few examples are known for bacterial predators. Bacteriovorax are halophilic prokaryotes that prey on susceptible gram-negative bacteria. We recently reported novel observations on the differential selection of Bacteriovorax phylotypes by two different prey, Vibrio parahaemolyticus and Vibrio vulnificus. However, the conclusion is restricted by the limited number of prey tested. In this study, we have conducted two independent investigations involving eight species of prey bacteria while using V. vulnificus and V. parahaemolytics as reference strains. Water samples collected from Dry Bar, Apalachicola Bay were used to establish microcosms which were respectively spiked with prey strains Vibrio cholerae, Escherichia coli or Pseudomonas putida to examine the response of native Bacteriovorax to freshwater bacteria. Indigenous Vibrio sp., Pseudoalteromonas sp., Photobacterium sp. and a clinical strain of V. vulnificus were also tested for the impact of saltwater prey on the Bacteriovorax community. At 24 hour intervals, optical density of the microcosm samples and the abundance of Bacteriovorax were measured over five days. The predominant Bacteriovorax plaques were selected and analyzed by 16S rRNA gene amplification and sequencing. In addition, the impacts of prey on predator population and bacterial community composition were investigated using culture independent denaturing gradient gel electrophoresis. Strikingly, Cluster IV was found consistently as the predominant phylotype produced by the freshwater prey. For all saltwater prey, subgroups of Bacteriovorax phylotype IX were the major predators recovered. The results suggest that prey is an important factor along with temperature, salinity and other environmental parameters in shaping Bacteriovorax communities in aquatic systems.     

Effects of surfactant adsorption and biodegradability on the distribution of bacteria between sediments and water in a freshwater microcosm.

A microcosm containing resuspended river sediment was used to investigate the effect of anionic surfactants on the distribution of bacteria between planktonic and attached populations. Freshwater river sediment containing viable bacteria was preequilibrated in the microcosm, which was subsequently supplemented with biodegradable or recalcitrant surfactants and a non-surface-active carbon and energy source. Population dynamics of both free-living and attached bacteria were measured by epifluorescence microscopy with simultaneous analysis of the residual solution concentration of the xenobiotic carbon source. The addition of the readily biodegradable anionic surfactants sodium decyl sulfate and sodium dodecyl sulfate in separate experiments caused an increase in the number of attached bacteria and a concomitant decrease in the number of free-living bacteria. As biodegradation of the surfactants progressed, these trends reversed and the bacterial populations had returned to their preaddition values by the time when biodegradation was completed. In contrast, sodium tetradecyl sulfate or sodium dodecane sulfonate did not stimulate bacterial association with sediment, nor were they biodegraded in the microcosm. Sodium pyruvate, a non-surface-active carbon and energy source, was readily utilized but caused no bacterial attachment to the sediment. These results indicate that for an anionic surfactant to induce bacterial attachment to river sediment, it must be biodegradable. The bacterial attachment to the sediment appears to be reversible and may be dependent on the accumulation of the surfactant at the surface or as a result of alteration of the surface free energies.     

Effects of surfactant adsorption and biodegradability on the distribution of bacteria between sediments and water in a freshwater microcosm.

A microcosm containing resuspended river sediment was used to investigate the effect of anionic surfactants on the distribution of bacteria between planktonic and attached populations. Freshwater river sediment containing viable bacteria was preequilibrated in the microcosm, which was subsequently supplemented with biodegradable or recalcitrant surfactants and a non-surface-active carbon and energy source. Population dynamics of both free-living and attached bacteria were measured by epifluorescence microscopy with simultaneous analysis of the residual solution concentration of the xenobiotic carbon source. The addition of the readily biodegradable anionic surfactants sodium decyl sulfate and sodium dodecyl sulfate in separate experiments caused an increase in the number of attached bacteria and a concomitant decrease in the number of free-living bacteria. As biodegradation of the surfactants progressed, these trends reversed and the bacterial populations had returned to their preaddition values by the time when biodegradation was completed. In contrast, sodium tetradecyl sulfate or sodium dodecane sulfonate did not stimulate bacterial association with sediment, nor were they biodegraded in the microcosm. Sodium pyruvate, a non-surface-active carbon and energy source, was readily utilized but caused no bacterial attachment to the sediment. These results indicate that for an anionic surfactant to induce bacterial attachment to river sediment, it must be biodegradable. The bacterial attachment to the sediment appears to be reversible and may be dependent on the accumulation of the surfactant at the surface or as a result of alteration of the surface free energies.     

A preliminary investigation of the effects of salinity on the bacterial flora of the Tay estuary.

Two features of the effects of salinity on the bacterial flora of the Tay estuary are considered. First the effect of salinity on the size and species composition of the resident microflora at various sites within the estuary. The second part is concerned with data obtained from laboratory experiments designed to study the effects of subjecting marine and freshwater bacteria, isolated from the Tay, to salt stress.     

Diversity and vertical distribution of magnetotactic bacteria along chemical gradients in freshwater microcosms

The vertical distribution of magnetotactic bacteria along various physico-chemical gradients in freshwater microcosms was analyzed by a combined approach of viable cell counts, 16S rRNA gene analysis, microsensor profiling and biogeochemical methods. The occurrence of magnetotactic bacteria was restricted to a narrow sediment layer overlapping or closely below the maximum oxygen and nitrate penetration depth. Different species showed different preferences within vertical gradients, but the largest proportion (63-98%) of magnetotactic bacteria was detected within the suboxic zone. In one microcosm the community of magnetotactic bacteria was dominated by one species of a coccoid "Alphaproteobacterium", as detected by denaturing gradient gel electrophoresis in sediment horizons from 1 to 10 mm depth. Maximum numbers of magnetotactic bacteria were up to 1.5 x 10(7) cells/cm3, which corresponded to 1% of the total cell number in the upper sediment layer. The occurrence of magnetotactic bacteria coincided with the availability of significant amounts (6-60 microM) of soluble Fe(II), and in one sample with hydrogen sulfide (up to 40 microM). Although various trends were clearly observed, a strict correlation between the distribution of magnetotactic bacteria and individual geochemical parameters was absent. This is discussed in terms of metabolic adaptation of various strains of magnetotactic bacteria to stratified sediments and diversity of the magnetotactic bacterial communities.     

Assembly processes and source tracking of planktonic and benthic bacterial communities in the Yellow River estuary

Estuaries connect rivers with the ocean and are considered transition regions due to the continuous inputs from rivers. Microbiota from different sources converge and undergo succession in these transition regions, but their assembly mechanisms along environmental gradients remain unclear. Here, we found that salinity had a stronger effect on planktonic than on benthic microbial communities, and the dominant planktonic bacteria changed more distinctly than the dominant benthic bacteria with changes in salinity. The planktonic bacteria in the brackish water came mainly from seawater, which was confirmed in the laboratory, whereas the benthic bacteria were weakly affected by salinity, which appeared to be a mixture of the bacteria from riverine and oceanic sediments. Benthic bacterial community assembly in the sediments was mainly controlled by homogeneous selection and almost unaffected by changes in salinity, the dominant assemblage processes for planktonic bacteria changed dramatically along the salinity gradient, from homogeneous selection in freshwater to drift in seawater. Our results highlight that salinity is the key driver of estuarine microbial succession and that salinity is more important in shaping planktonic than benthic bacterial communities in the Yellow River estuary.     

Testing for beneficial reversal of dominance during salinity shifts in the invasive copepod Eurytemora affinis,and implications for the maintenance of genetic variation

Maintenance of genetic variation at loci under selection has profound implications for adaptation under environmental change. In temporally and spatially varying habitats, non‐neutral polymorphism could be maintained by heterozygote advantage across environments (marginal overdominance), which could be greatly increased by beneficial reversal of dominance across conditions. We tested for reversal of dominance and marginal overdominance in salinity tolerance in the saltwater‐to‐freshwater invading copepod Eurytemora affinis. We compared survival of F1 offspring generated by crossing saline and freshwater inbred lines (between‐salinity F1 crosses) relative to within‐salinity F1 crosses, across three salinities. We found evidence for both beneficial reversal of dominance and marginal overdominance in salinity tolerance. In support of reversal of dominance, survival of between‐salinity F1 crosses was not different from that of freshwater F1 crosses under freshwater conditions and saltwater F1 crosses under saltwater conditions. In support of marginal overdominance, between‐salinity F1 crosses exhibited significantly higher survival across salinities relative to both freshwater and saltwater F1 crosses. Our study provides a rare empirical example of complete beneficial reversal of dominance associated with environmental change. This mechanism might be crucial for maintaining genetic variation in salinity tolerance in E. affinis populations, allowing rapid adaptation to salinity changes during habitat invasions.     

Effects of Deposit-Feeding Macrofauna on Benthic Bacteria,Viruses, and Protozoa in a Silty Freshwater Sediment

In microcosm experiments, we simultaneously tested the effects of increased numbers of deposit-feeding macrofauna (chironomids, oligochaetes and cladocerans) on the standing stock, activities and interactions of heterotrophic bacteria, viruses, and bacterivorous protozoa (heterotrophic nanoflagellates and ciliates) in the aerobic layer of a silty littoral freshwater sediment. On average, bacterial secondary production was stimulated between 11 and 29% by all macrofaunal groups compared to control experiments without macrofauna addition. Bacterial standing stock increased significantly by 8 and 13% in case of chironomids and cladocerans, respectively. Oligochaetes and chironomids produced significant negative effects on viral abundance while the results with cladocerans were inconsistent. The addition of oligochaetes and chironomids resulted in a significant decrease by on average 68 and 32% of viral decay rates, respectively, used as a measure of viral production. The calculated contribution of virus-induced lysis to benthic bacterial mortality was low, with 2.8 to 11.8% of bacterial secondary production, and decreased by 39 to 81% after the addition of macrofauna compared to the control. The abundances of heterotrophic nanoflagellates were significantly reduced by 20% by all tested macrofauna groups, while ciliates showed inconsistent results. The importance of heterotrophic nanoflagellate grazing on benthic bacteria was very low (<1% of bacterial secondary production) and was further reduced by elevated numbers of macrofauna. Thus, the selected deposit feeding macrofauna groups seem to have several direct and indirect and partly antagonistic effects on the benthic bacterial compartment through the enhancement of bacterial production and the reduction of virus-induced cell lysis and protozoan grazing.     

Macrophyte species drive the variation of bacterioplankton community composition in a shallow freshwater lake

Macrophytes play an important role in structuring aquatic ecosystems. In this study, we explored whether macrophyte species are involved in determining the bacterioplankton community composition (BCC) in shallow freshwater lakes. The BCC in field areas dominated by different macrophyte species in Taihu Lake, a large, shallow freshwater lake, was investigated over a 1-year period. Subsequently, microcosm experiments were conducted to determine if single species of different types of macrophytes in an isolated environment would alter the BCC. Denaturing gradient gel electrophoresis (DGGE), followed by cloning and sequence analysis of selected samples, was employed to analyze the BCC. The DGGE results of the field investigations indicated that the BCC changed significantly from season to season and that the presence of different macrophyte species resulted in lower BCC similarities in the summer and fall. LIBSHUFF analysis of selected clone libraries from the summer demonstrated different BCCs in the water column surrounding different macrophytes. Relative to the field observations, the microcosm studies indicated that the BCC differed more pronouncedly when associated with different species of macrophytes, which was also supported by LIBSHUFF analysis of the selected clone libraries. Overall, this study suggested that macrophyte species might be an important factor in determining the composition of bacterial communities in this shallow freshwater lake and that the species-specific influence of macrophytes on BCC is variable with the season and distance.     

Changes in Community Structure of Sediment Bacteria Along the Florida Coastal Everglades Marsh–Mangrove–Seagrass Salinity Gradient

Community structure of sediment bacteria in the Everglades freshwater marsh, fringing mangrove forest, and Florida Bay seagrass meadows were described based on polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) patterns of 16S rRNA gene fragments and by sequencing analysis of DGGE bands. The DGGE patterns were correlated with the environmental variables by means of canonical correspondence analysis. There was no significant trend in the Shannon–Weiner index among the sediment samples along the salinity gradient. However, cluster analysis based on DGGE patterns revealed that the bacterial community structure differed according to sites. Not only were these salinity/vegetation regions distinct but the sediment bacteria communities were consistently different along the gradient from freshwater marsh, mangrove forest, eastern-central Florida Bay, and western Florida Bay. Actinobacteria- and Bacteroidetes/Chlorobi-like DNA sequences were amplified throughout all sampling sites. More Chloroflexi and members of candidate division WS3 were found in freshwater marsh and mangrove forest sites than in seagrass sites. The appearance of candidate division OP8-like DNA sequences in mangrove sites distinguished these communities from those of freshwater marsh. The seagrass sites were characterized by reduced presence of bands belonging to Chloroflexi with increased presence of those bands related to Cyanobacteria, γ-Proteobacteria, Spirochetes, and Planctomycetes. This included the sulfate-reducing bacteria, which are prevalent in marine environments. Clearly, bacterial communities in the sediment were different along the gradient, which can be explained mainly by the differences in salinity and total phosphorus.