Bacterial Community Dynamics and Polycyclic Aromatic Hydrocarbon Degradation during Bioremediation of Heavily Creosote-Contaminated Soil

Bacterial community dynamics and biodegradation processes were examined in a highly creosote-contaminated soil undergoing a range of laboratory-based bioremediation treatments. The dynamics of the eubacterial community, the number of heterotrophs and polycyclic aromatic hydrocarbon (PAH) degraders, and the total petroleum hydrocarbon (TPH) and PAH concentrations were monitored during the bioremediation process. TPH and PAHs were significantly degraded in all treatments (72 to 79% and 83 to 87%, respectively), and the biodegradation values were higher when nutrients were not added, especially for benzo(a)anthracene and chrysene. The moisture content and aeration were determined to be the key factors associated with PAH bioremediation. Neither biosurfactant addition, bioaugmentation, nor ferric octate addition led to differences in PAH or TPH biodegradation compared to biodegradation with nutrient treatment. All treatments resulted in a high first-order degradation rate during the first 45 days, which was markedly reduced after 90 days. A sharp increase in the size of the heterotrophic and PAH-degrading microbial populations was observed, which coincided with the highest rates of TPH and PAH biodegradation. At the end of the incubation period, PAH degraders were more prevalent in samples to which nutrients had not been added. Denaturing gradient gel electrophoresis analysis and principal-component analysis confirmed that there was a remarkable shift in the composition of the bacterial community due to both the biodegradation process and the addition of nutrients. At early stages of biodegradation, the α-Proteobacteria group (genera Sphingomonas and Azospirillum) was the dominant group in all treatments. At later stages, the γ-Proteobacteria group (genus Xanthomonas), the α-Proteobacteria group (genus Sphingomonas), and the Cytophaga-Flexibacter-Bacteroides group (Bacteroidetes) were the dominant groups in the nonnutrient treatment, while the γ-Proteobacteria group (genus Xathomonas), the β-Proteobacteria group (genera Alcaligenes and Achromobacter), and the α-Proteobacteria group (genus Sphingomonas) were the dominant groups in the nutrient treatment. This study shows that specific bacterial phylotypes are associated both with different phases of PAH degradation and with nutrient addition in a preadapted PAH-contaminated soil. Our findings also suggest that there are complex interactions between bacterial species and medium conditions that influence the biodegradation capacity of the microbial communities involved in bioremediation processes.     

Microbial Population Changes during Bioremediation of an Experimental Oil Spill

Three crude oil bioremediation techniques were applied in a randomized block field experiment simulating a coastal oil spill. Four treatments (no oil control, oil alone, oil plus nutrients, and oil plus nutrients plus an indigenous inoculum) were applied. In situ microbial community structures were monitored by phospholipid fatty acid (PLFA) analysis and 16S rDNA PCR-denaturing gradient gel electrophoresis (DGGE) to (i) identify the bacterial community members responsible for the decontamination of the site and (ii) define an end point for the removal of the hydrocarbon substrate. The results of PLFA analysis demonstrated a community shift in all plots from primarily eukaryotic biomass to gram-negative bacterial biomass with time. PLFA profiles from the oiled plots suggested increased gram-negative biomass and adaptation to metabolic stress compared to unoiled controls. DGGE analysis of untreated control plots revealed a simple, dynamic dominant population structure throughout the experiment. This banding pattern disappeared in all oiled plots, indicating that the structure and diversity of the dominant bacterial community changed substantially. No consistent differences were detected between nutrient-amended and indigenous inoculum-treated plots, but both differed from the oil-only plots. Prominent bands were excised for sequence analysis and indicated that oil treatment encouraged the growth of gram-negative microorganisms within the α-proteobacteria and Flexibacter-Cytophaga-Bacteroides phylum. α-Proteobacteria were never detected in unoiled controls. PLFA analysis indicated that by week 14 the microbial community structures of the oiled plots were becoming similar to those of the unoiled controls from the same time point, but DGGE analysis suggested that major differences in the bacterial communities remained.     

Bacterial Community Dynamics during Bioremediation of Diesel Oil-Contaminated Antarctic Soil

The effect of nutrient and inocula amendment in a bioremediation field trial using a nutrient-poor Antarctic soil chronically contaminated with hydrocarbons was tested. The analysis of the effects that the treatments caused in bacterial numbers and hydrocarbon removal was combined with the elucidation of the changes occurring on the bacterial community, by 16S rDNA-based terminal restriction fragment length polymorphism (T-RFLP) typing, and the detection of some of the genes involved in the catabolism of hydrocarbons. All treatments caused a significant increase in the number of bacteria able to grow on hydrocarbons and a significant decrease in the soil hydrocarbon content, as compared to the control. However, there were no significant differences between treatments. Comparison of the soil T-RFLP profiles indicated that there were changes in the structure and composition of bacterial communities during the bioremediation trial, although the communities in treated plots were highly similar irrespective of the treatment applied, and they had a similar temporal dynamics. These results showed that nutrient addition was the main factor contributing to the outcome of the bioremediation experiment. This was supported by the lack of evidence of the establishment of inoculated consortia in soils, since their characteristic electrophoretic peaks were only detectable in soil profiles at the beginning of the experiment. Genetic potential for naphthalene degradation, evidenced by detection of nahAc gene, was observed in all soil plots including the control. In treated plots, an increase in the detection of catechol degradation genes (nahH and catA) and in a key gene of denitrification (nosZ) was observed as well. These results indicate that treatments favored the degradation of aromatic hydrocarbons and probably stimulated denitrification, at least transiently. This mesocosm study shows that recovery of chronically contaminated Antarctic soils can be successfully accelerated using biostimulation with nutrients, and that this causes a change in the indigenous bacterial communities and in the genetic potential for hydrocarbon degradation. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Bioremediation of Organometallic Compounds by Bacterial Degradation

The use of organometallic compounds in the environment is constantly increasing with increased technology and progress in scientific research. But since these compounds are fairly stable, as metallic bonds are stable, they are difficult to degrade or decompose naturally. The aim of this work was to isolate and characterize heterotrophic bacteria that can degrade organometallic compounds (in this case ‘ferrocene’ and its derivatives). A Gram-negative coccobacillus was isolated from a rusting iron pipe draining into a freshwater lake, which could utilize ferrocene as a sole source of carbon. Phylogenetic analysis based on 16S rDNA sequence suggested that the isolated organism resembled an environmental isolate of Bordetella. Ferrocene degradation was confirmed by plotting the growth curve of the bacterium in a medium with ferrocene as the sole source of carbon. Further confirmation of degradation of ferrocene and its derivatives was done using Gas Chromatography Mass Spectroscopy. Since the bacterium degraded organometallic compounds and released the metal in liquid medium, it could be suggested that this organism can also be used for extracting metal ions from organo-metal containing wastes.     

石油污染海域的微生物群落及烃的降解

目前世界海洋石油污染问题已经严重威胁到海洋生态环境的安全,生物修复是一种处理石油污染的新方法。综述了海洋环境中烃降解微生物生态学方面的一些研究进展,包括探测未培养细菌的新方法、新的分离方法及主要的烃降解菌株的特性,以便努力改进现有分离石油降解菌及石油乳化细菌的方法,同时发现对于石油降解有益的新菌种。     

Shoreline Bioremediation Following the Exxon Valdez Oil Spill in Alaska

Bioremediation played an important role in the cleanup following the oil spill from the Exxon Valdez in Prince William Sound, Alaska. The initial spill response included washing the beaches and collecting the oil with skimmers, but while this was in progress a joint program between Exxon and the United States Environmental Protection Agency investigated the feasibility of using bioremediation to remove residual oil. Early experiments demonstrated that oil biodegradation on affected shorelines was nutrient-limited, and several fertilizer application strategies were tested to overcome this limitation. Two fertilizers, the liquid oleophilic Inipol EAP22 and the solid slow-release Customblen™, were chosen for wide-scale application. Field monitoring convincingly demonstrated that the fertilizer strategy effectively stimulated oil biodegradation severalfold, with no adverse environmental impacts.     

From Rare to Dominant: a Fine-Tuned Soil Bacterial Bloom during Petroleum Hydrocarbon Bioremediation

Hydrocarbons are worldwide-distributed pollutants that disturb various ecosystems. The aim of this study was to characterize the short-lapse dynamics of soil microbial communities in response to hydrocarbon pollution and different bioremediation treatments. Replicate diesel-spiked soil microcosms were inoculated with either a defined bacterial consortium or a hydrocarbonoclastic bacterial enrichment and incubated for 12 weeks. The microbial community dynamics was followed weekly in microcosms using Illumina 16S rRNA gene sequencing. Both the bacterial consortium and enrichment enhanced hydrocarbon degradation in diesel-polluted soils. A pronounced and rapid bloom of a native gammaproteobacterium was observed in all diesel-polluted soils. A unique operational taxonomic unit (OTU) related to the Alkanindiges genus represented ∼0.1% of the sequences in the original community but surprisingly reached >60% after 6 weeks. Despite this Alkanindiges-related bloom, inoculated strains were maintained in the community and may explain the differences in hydrocarbon degradation. This study shows the detailed dynamics of a soil bacterial bloom in response to hydrocarbon pollution, resembling microbial blooms observed in marine environments. Rare community members presumably act as a reservoir of ecological functions in high-diversity environments, such as soils. This rare-to-dominant bacterial shift illustrates the potential role of a rare biosphere facing drastic environmental disturbances. Additionally, it supports the concept of “conditionally rare taxa,” in which rareness is a temporary state conditioned by environmental constraints.     

Effects of Spilled Oil on Bacterial Communities of Mediterranean Coastal Anoxic Sediments Chronically Subjected to Oil Hydrocarbon Contamination

The effects of spilled oil on sedimentary bacterial communities were examined in situ at 20 m water depth in a Mediterranean coastal area. Sediment collected at an experimental site chronically subjected to hydrocarbon inputs was reworked into PVC cores with or without a massive addition of crude Arabian light oil (∼20 g kg⁻¹ dry weight). Cores were reinserted into the sediment and incubated in situ at the sampling site (20 m water depth) for 135 and 503 days. The massive oil contamination induced significant shifts in the structure of the indigenous bacterial communities as shown by ribosomal intergenic spacer analysis (RISA). The vertical heterogeneity of the bacterial communities within the sediment was more pronounced in the oiled sediments particularly after 503 days of incubation. Response to oil of the deeper depth communities (8–10 cm) was slower than that of superficial depth communities (0–1 and 2–4 cm). Analysis of the oil composition by gas chromatography revealed a typical microbial alteration of n-alkanes during the experiment. Predominant RISA bands in oiled sediments were affiliated to hydrocarbonoclastic bacteria sequences. In particular, a 395-bp RISA band, which was the dominant band in all the oiled sediments for both incubation times, was closely related to hydrocarbonoclastic sulfate-reducing bacteria (SRB). These bacteria may have contributed to the main fingerprint changes and to the observed biodegradation of n-alkanes. This study provides useful information on bacterial dynamics in anoxic contaminated infralittoral sediments and highlights the need to assess more precisely the contribution of SRB to bioremediation in oil anoxic contaminated areas.     

Kinetics of Polycyclic Aromatic Hydrocarbon (PAH) Degradation in Long-term Polluted Soils during Bioremediation

Bioremediation experiments with ten different soil samples from former industrial sites which were long-term polluted with polycyclic aromatic hydrocarbons (PAHs) were carried out using outdoor pot trials. The degradation of 15 PAHs according to the US EPA was investigated for 168 weeks through repeated soil sampling and determination of the total PAH concentration. On average, degradation was largest for acenaphthene (88%; 63 to 99%) and smallest for anthracene (22%; no significant degradation to 89%). For most of the PAH single substances, degradation kinetics were characterised by a first initial phase of fast degradation. In a subsequent second phase, degradation diminished and residual PAH concentrations were approached within 168 weeks, resulting in a similar PAH pattern in the ten soil samples. Degradation kinetics was calculated through the selection of the appropriate differential rate equation from a set of seven equations. Kinetics of PAH degradation was best fitted by single and two coupled first order exponential equations with median R2 of 0.71 (0.01 to 1.00). Degradation rate constants of the rapid phase (k ₁) ranged from 0.05×10⁻² week⁻¹ for benzo[k]fluoranthene to 18.3 week⁻¹ for naphthalene and for the subsequent slow degradation phase (k ₂) they ranged from 0.01×10⁻² week⁻¹ for benzo[a]anthracene to 2.3×10⁻² week⁻¹ for fluoranthene. Degradation was governed by desorption and diffusion processes of different rates, while microbial activity did not influence the kinetics. Median disappearance times (DT₅₀) ranged from 6.1 weeks for naphthalene to 522 weeks for benzo[k]fluoranthene. With the exception of the 6-ring PAHs dibenzo[ah]anthracene and indeno[1,2,3-cd]pyrene, this sequence followed the PAHs’ degree of condensation. The total initial PAH concentration and the residual concentration were correlated with R2 of 0.69, with larger initial PAH concentrations leading to larger residual concentrations and degradation rates.     

双孢蘑菇培养料发酵过程中细菌群落动态变化

双孢蘑菇产量的最主要影响因素是培养料的堆肥发酵过程,而微生物是其中的主要推动力。为了揭示工厂化生产双孢蘑菇堆肥发酵过程中的微生物群落结构变化及演替规律,深入了解细菌在堆肥发酵过程中作用,文章以工厂化生产双孢蘑菇堆肥为研究对象,采用16S rRNA基因高通量测序技术分析堆肥前、一次发酵、二次发酵过程中细菌群落特征。共产生有效操作分类单元(OTUs) 854个,涵盖了19门259属的细菌。堆肥前优势类群为Proteobacteria,一次发酵优势类群为Firmicutes、Deinococcus-Thermus等,二次发酵时绿弯菌门Chloroflexi为主要类群。研究结果表明:微生物种类和数量随堆肥过程不断升高,并在二次发酵后降低,且微生物群落结构呈现连续变化的规律。高通量测序还发现了很多未分类细菌,说明双孢蘑菇培养料堆肥样品中还蕴含着大量未知的微生物种类。     

Bacterial Communities from Shoreline Environments (Costa da Morte,Northwestern Spain) Affected by the Prestige Oil Spill

The bacterial communities in two different shoreline matrices, rocks and sand, from the Costa da Morte, northwestern Spain, were investigated 12 months after being affected by the Prestige oil spill. Culture-based and culture-independent approaches were used to compare the bacterial diversity present in these environments with that at a nonoiled site. A long-term effect of fuel on the microbial communities in the oiled sand and rock was suggested by the higher proportion of alkane and polyaromatic hydrocarbon (PAH) degraders and the differences in denaturing gradient gel electrophoresis patterns compared with those of the reference site. Members of the classes Alphaproteobacteria and Actinobacteria were the prevailing groups of bacteria detected in both matrices, although the sand bacterial community exhibited higher species richness than the rock bacterial community did. Culture-dependent and -independent approaches suggested that the genus Rhodococcus could play a key role in the in situ degradation of the alkane fraction of the Prestige fuel together with other members of the suborder Corynebacterineae. Moreover, other members of this suborder, such as Mycobacterium spp., together with Sphingomonadaceae bacteria (mainly Lutibacterium anuloederans), were related as well to the degradation of the aromatic fraction of the Prestige fuel. The multiapproach methodology applied in the present study allowed us to assess the complexity of autochthonous microbial communities related to the degradation of heavy fuel from the Prestige and to isolate some of their components for a further physiological study. Since several Corynebacterineae members related to the degradation of alkanes and PAHs were frequently detected in this and other supralittoral environments affected by the Prestige oil spill along the northwestern Spanish coast, the addition of mycolic acids to bioremediation amendments is proposed to favor the presence of these degraders in long-term fuel pollution-affected areas with similar characteristics.Since the Polycommander accident, many other oil spills, such as the Urquiola (1976), Andros Patria (1978), and Aegean Sea (1992) spills, have occurred on the Galician coast (northwestern Spain), where intense maritime traffic takes place. On 13 November 2002, the oil tanker Prestige sprang a leak off Cape Finisterre (Galicia, northwestern Spain) and 6 days later its oil tank broke up and sank 240 km west of Galicia. The spill of 60,000 tons of heavy fuel oil polluted 500 miles of the Spanish coast, reaching the French coast. The Costa da Morte, northwestern Spain, was the most affected area (2). The oil residue released by the Prestige was devoid of the more labile fractions (boiling point, <300°C), with high levels of aromatic hydrocarbons (∼50%), as well as resins and asphaltenes (∼30%) (6).Information about the autochthonous microbial populations at maritime oil-polluted sites is scarce (23). The studies carried out after the Nakhodka spilled oil with a chemical composition similar to that of the Prestige fuel, gathered information on the marine microbial populations that adapted to heavy fuel oil. Different molecular approaches, mainly involving 16S rRNA gene analyses such as PCR-denaturing gradient gel electrophoresis (DGGE) (31), clone libraries, and specific oligonucleotide probes (43), were used to describe the bacterial communities established in different environments and at different time intervals after the oil spill.Most of the previous studies were focused either on the isolation of a few culturable degrading strains or just on detecting the 16S rRNA gene sequences of all of the bacteria present in polluted samples without gathering information on their physiology. As a consequence, more efforts should be made to understand community structures in situ and to isolate the key oil-degrading species present, with the aim to further investigate their requirements (23, 58) that could be used in the development of new bioremediation.In the present report, we describe a microbiological analysis of a cobblestone beach on the Costa da Morte, northwestern Spain, affected by the Prestige heavy fuel oil spill 12 months after the last fuel stranding. The microbial community was examined thoroughly by a triple-approach method based on different cultivation strategies and culture-independent methods such as DGGE and the screening of 16S rRNA gene clone libraries.     

Response of Archaeal Communities in Beach Sediments to Spilled Oil and Bioremediation

While the contribution of Bacteria to bioremediation of oil-contaminated shorelines is well established, the response of Archaea to spilled oil and bioremediation treatments is unknown. The relationship between archaeal community structure and oil spill bioremediation was examined in laboratory microcosms and in a bioremediation field trial. 16S rRNA gene-based PCR and denaturing gradient gel analysis revealed that the archaeal community in oil-free laboratory microcosms was stable for 26 days. In contrast, in oil-polluted microcosms a dramatic decrease in the ability to detect Archaea was observed, and it was not possible to amplify fragments of archaeal 16S rRNA genes from samples taken from microcosms treated with oil. This was the case irrespective of whether a bioremediation treatment (addition of inorganic nutrients) was applied. Since rapid oil biodegradation occurred in nutrient-treated microcosms, we concluded that Archaea are unlikely to play a role in oil degradation in beach ecosystems. A clear-cut relationship between the presence of oil and the absence of Archaea was not apparent in the field experiment. This may have been related to continuous inoculation of beach sediments in the field with Archaea from seawater or invertebrates and shows that the reestablishment of Archaea following bioremediation cannot be used as a determinant of ecosystem recovery following bioremediation. Comparative 16S rRNA sequence analysis showed that the majority of the Archaea detected (94%) belonged to a novel, distinct cluster of group II uncultured Euryarchaeota, which exhibited less than 87% identity to previously described sequences. A minor contribution of group I uncultured Crenarchaeota was observed.     

Role of Bacterial Exopolysaccharides (EPS) in the Fate of the Oil Released during the Deepwater Horizon Oil Spill

Halomonas species are recognized for producing exopolysaccharides (EPS) exhibiting amphiphilic properties that allow these macromolecules to interface with hydrophobic substrates, such as hydrocarbons. There remains a paucity of knowledge, however, on the potential of Halomonas EPS to influence the biodegradation of hydrocarbons. In this study, the well-characterized amphiphilic EPS produced by Halomonas species strain TG39 was shown to effectively increase the solubilization of aromatic hydrocarbons and enhance their biodegradation by an indigenous microbial community from oil-contaminated surface waters collected during the active phase of the Deepwater Horizon oil spill. Three Halomonas strains were isolated from the Deepwater Horizon site, all of which produced EPS with excellent emulsifying qualities and shared high (97-100%) 16S rRNA sequence identity with strain TG39 and other EPS-producing Halomonas strains. Analysis of pyrosequence data from surface water samples collected during the spill revealed several distinct Halomonas phylotypes, of which some shared a high sequence identity (≥97%) to strain TG39 and the Gulf spill isolates. Other bacterial groups comprising members with well-characterized EPS-producing qualities, such as Alteromonas , Colwellia and Pseudoalteromonas , were also found enriched in surface waters, suggesting that the total pool of EPS in the Gulf during the spill may have been supplemented by these organisms. Roller bottle incubations with one of the Halomonas isolates from the Deepwater Horizon spill site demonstrated its ability to effectively produce oil aggregates and emulsify the oil. The enrichment of EPS-producing bacteria during the spill coupled with their capacity to produce amphiphilic EPS is likely to have contributed to the ultimate removal of the oil and to the formation of oil aggregates, which were a dominant feature observed in contaminated surface waters.     

石油集输系统中微生物群落结构研究

采用16SrRNA基因克隆一变性梯度凝胶电泳分析方法研究了石油集输系统原油和油田产水中的微生物群落结构。变性梯度凝胶电泳图谱显示：油田产水中微生物群落远比原油中的菌群丰富。所有的油田水样和原油样本中都存在与Ochrobactrum sp．和Stenotrophomonas sp．相关的细菌;原油样本中检测出与Burkholderia sp．、Brevundimonas sp．和Propionibacterium sp．相关的细菌,而这些细菌在油田水样中未检出;在油田水样中检出与Hippea sp．、Acidovorax sp．、Arcobacter sp．、Pseudomonas sp．、Thiomicrospira sp．、Brevibacterium sp．、Tissierella sp．和Peptostreptococcus sp．相关的细菌,而这些细菌在原油样本中未检出。用古细菌特异性引物进行检测发现在油田水样中存在与Methanomicrobials和Methanosarcinales相关的产甲烷菌,而这些细菌在原油样本中未检出。在石油集输过程中,油田水样和原油中微生物群落的相似性分别为83．3％和88．2％,说明微生物群落结构较为稳定。     

Protozoa Drive the Dynamics of Culturable Biocontrol Bacterial Communities

Some soil bacteria protect plants against soil-borne diseases by producing toxic secondary metabolites. Such beneficial biocontrol bacteria can be used in agricultural systems as alternative to agrochemicals. The broad spectrum toxins responsible for plant protection also inhibit predation by protozoa and nematodes, the main consumers of bacteria in soil. Therefore, predation pressure may favour biocontrol bacteria and contribute to plant health. We analyzed the effect of Acanthamoeba castellanii on semi-natural soil bacterial communities in a microcosm experiment. We determined the frequency of culturable bacteria carrying genes responsible for the production of the antifungal compounds 2,4-diacetylphloroglucinol (DAPG), pyrrolnitrin (PRN) and hydrogen cyanide (HCN) in presence and absence of A. castellanii. We then measured if amoebae affected soil suppressiveness in a bioassay with sugar beet seedlings confronted to the fungal pathogen Rhizoctonia solani. Amoebae increased the frequency of both DAPG and HCN positive bacteria in later plant growth phases (2 and 3 weeks), as well as the average number of biocontrol genes per bacterium. The abundance of DAPG positive bacteria correlated with disease suppression, suggesting that their promotion by amoebae may enhance soil health. However, the net effect of amoebae on soil suppressiveness was neutral to slightly negative, possibly because amoebae slow down the establishment of biocontrol bacteria on the recently emerged seedlings used in the assay. The results indicate that microfaunal predators foster biocontrol bacterial communities. Understanding interactions between biocontrol bacteria and their predators may thus help developing environmentally friendly management practices of agricultural systems.     

Dynamics of Seawater Bacterial Communities in a Shellfish Hatchery

Bacterial disease is a significant issue for larviculture of several species of shellfish, including oysters. One source of bacteria is the seawater used throughout the hatchery. In this study carried out at a commercial oyster hatchery in Tasmania, Australia, the diversity of the bacterial community and its relationship with larval production outcomes were studied over a 2-year period using terminal restriction fragment length polymorphism and tag-encoded pyrosequencing. The bacterial communities were very diverse, dominated by the Alphaproteobacteria, Gammaproteobacteria, Flavobacteria and Cyanobacteria. The communities were highly variable on scales of days, weeks and seasons. The difference between the intake seawater and treated clean seawater used in the hatchery was smaller than the observed temporal differences in the seawater throughout the year. No clear correlation was observed between production outcomes and the overall bacterial community structure. However, one group of Cyanobacterial sequences was more abundant when mass mortality events occurred than when healthy spat were produced although they were always present.     

Bacterial Communities Associated with Culex Mosquito Larvae and Two Emergent Aquatic Plants of Bioremediation Importance

Microbes are important for mosquito nutrition, growth, reproduction and control. In this study, we examined bacterial communities associated with larval mosquitoes and their habitats. Specifically, we characterized bacterial communities associated with late larval instars of the western encephalitis mosquito (Culextarsalis), the submerged portions of two emergent macrophytes (California bulrush, Schoenoplectuscalifornicus and alkali bulrush, Schoenoplectusmaritimus), and the associated water columns to investigate potential differential use of resources by mosquitoes in different wetland habitats. Using next-generation sequence data from 16S rRNA gene hypervariable regions, the alpha diversity of mosquito gut microbial communities did not differ between pond mesocosms containing distinct monotypic plants. Proteobacteria, dominated by the genus Thorsellia (Enterobacteriaceae), was the most abundant phylum recovered from C. tarsalis larvae. Approximately 49% of bacterial OTUs found in larval mosquitoes were identical to OTUs recovered from the water column and submerged portions of the two bulrushes. Plant and water samples were similar to one another, both being dominated by Actinobacteria, Bacteroidetes, Cyanobacteria, Proteobacteria and Verrucomicrobia phyla. Overall, the bacterial communities within C. tarsalis larvae were conserved and did not change across sampling dates and between two distinct plant habitats. Although Thorsellia spp. dominated mosquito gut communities, overlap of mosquito gut, plant and water-column OTUs likely reveal the effects of larval feeding. Future research will investigate the role of the key indicator groups of bacteria across the different developmental stages of this mosquito species.