Rhodocyclaceae Family Research Articles

Integrated genomics provides insights into the evolution of the polyphosphate accumulation trait of Ca. Accumulibacter

Candidatus Accumulibacter, a prominent polyphosphate-accumulating organism (PAO) in wastewater treatment, plays a crucial role in enhanced biological phosphorus removal (EBPR). The genetic underpinnings of its polyphosphate accumulation capabilities, however, remain largely unknown. Here, we conducted a comprehensive genomic analysis of Ca. Accumulibacter-PAOs and their relatives within the Rhodocyclaceae family, identifying 124 core genes acquired via horizontal gene transfer (HGT) at its least common ancestor. Metatranscriptomic analysis of an enrichment culture of Ca. Accumulibacter revealed active transcription of 44 of these genes during an EBPR cycle, notably including the polyphosphate kinase 2 (PPK2) gene instead of the commonly recognized polyphosphate kinase 1 (PPK1) gene. Intriguingly, the phosphate regulon (Pho) genes showed minimal transcriptions, pointing to a distinctive fact of Pho dysregulation, where PhoU, the phosphate signaling complex protein, was not regulating the high-affinity phosphate transport (Pst) system, resulting in continuous phosphate uptake. To prevent phosphate toxicity, Ca. Accumulibacter utilized the laterally acquired PPK2 to condense phosphate into polyphosphate, resulting in the polyphosphate-accumulating feature. This study provides novel insights into the evolutionary emergence of the polyphosphate-accumulating trait in Ca. Accumulibacter, offering potential advancements in understanding the PAO phenotype in the EBPR process.

Identification of Antibiotic Resistance Gene Hosts in Treatment Wetlands Using a Single-Cell Based High-Throughput Approach

Determining the prevalence of antimicrobial resistance (AMR) in non-clinical settings is vital for better management of the global AMR crisis. Untreated and even treated wastewaters are important sources that release AMR into the environment. Methodologically, it is difficult to generate a comprehensive in situ profile of antibiotic resistance gene hosts. Here, we used epicPCR (emulsion, paired isolation, and concatenation PCR) as a cultivation-independent method to reveal the host profiles of the AMR indicator genes intI1, sul1, sul2, and dfrA1 in two constructed wetlands treating municipal wastewater. Overall, the epicPCR analysis revealed a profile of AMR indicator gene hosts that is consistent with literature data from cultivation-based approaches. Most carriers of antibiotic resistance (AR) genes and likely of class 1 integrons belonged to the Gammaproteobateria, particularly the Burkholderiaceae and Rhodocyclaceae families, followed by members of the Campylobacterota, Desulfobacterota, and Firmicutes. The analysis also identified several novel hosts for the indicator genes widely distributed in the wetlands, including the genera Legionella and Ralstonia. Therefore, the application of epicPCR has produced an expanded insight into the in situ indicator gene host profile, while highlighting the role of the environment as a reservoir for AMR.

Bacterial Communities in a Gradient of Abiotic Factors Near a Sulfide Thermal Spring in Northern Baikal

The structure and diversity of microbial communities developing in the combined gradient of temperature (44–19 °C), as well as concentration of oxygen (0–10 mg/L) and hydrogen sulfide (33–0.7 mg/L), were studied in the thermal sulfide spring on the coast of Northern Lake Baikal. The predominance of bacteria participating in sulfur and nitrogen cycles and significant changes in the composition of microbial communities were noted at changing physicochemical conditions. Thiovirga sp. (sulfur-oxidizing bacteria, up to 37%) and Azonexus sp. (nitrogen-fixing bacteria, up to 43%) were dominant at high temperatures and concentrations of hydrogen sulfide in two hydrotherms. In addition, a significant contribution of the Rhodocyclaceae family (up to 51%) which is involved in the denitrification processes, and Acetoanaerobium sp. (up to 20%) fixing carbon oxide were found in the spring water. In the stream, mainly oxygenic cyanobacteria (up to 56%) developed at a temperature of 33 °C, in the presence of hydrogen sulfide and oxygen. In addition, sulfur bacteria of the genus Thiothrix (up to 48%) found in epibiotic communities of benthic animals of Lake Baikal were present here. Thiothrix sp. formed massive fouling in the zone of mixing lake and thermal waters with a significant contribution of hydrogen-oxidizing bacteria of the genus Hydrogenophaga (up to 22.5%). As well as chemolitho- and phototrophic bacteria, chemoorganotrophs (phyla Firmicutes, Chloroflexi, Desulfobacterota, Nitrospirota, Fibrobacterota, etc.) have been identified in all communities. The chemical parameters of water in spring and coastal zones indicate a significant change in the composition of thermal waters occurring with the participation of diverse microbial communities that contribute to the assimilation of inorganic components of mineral thermal waters.

Hacking biofilm developed in a structured-bed reactor (SBRRIA) with integrated processes of nitrogen and organic matter removal.

Biomass samples from a structured-bed reactor subjected to recirculation and intermittent aeration (SBRRIA) were analyzed to investigate the bacterial community shift along with the changes in the C/N ratio. The C/N ratios tested were 7.6 ± 1.0 (LNC) and 2.9 ± 0.4 (HNC). The massive sequencing analyses revealed that the microbial community adjusted itself to different organic and nitrogenous applied loads, with no harm to reactor performance regarding COD and Total-N removal. Under LNC, conventional nitrification and heterotrophic denitrification steered the process, as indicated by the detection of microorganisms affiliated with Nitrosomonadaceae, Nitrospiraceae, and Rhodocyclaceae families. However, under HNC, the C/N ratio strongly affected the microbial community, resulting in the prevalence of members of Saprospiraceae, Chitinophagaceae, Xanthomonadaceae, Comamonadaceae, Bacillaceae, and Planctomycetaceae. These families include bacteria capable of using organic matter derived from cell lysis, ammonia-oxidizers under low DO, heterotrophic nitrifiers-aerobic denitrifiers, and non-isolated strains of Anammox. The DO profile confirmed that the stratification in aerobic, anoxic, and anaerobic zones enabled the establishment of different nitrogen degradation pathways, including the Anammox.

Alkylphenols and Chlorophenols Remediation in Vertical Flow Constructed Wetlands: Removal Efficiency and Microbial Community Response

This study aims to investigate the effect of two different groups of phenolic compounds (the alkylphenols nonylphenol (NP) and octylphenol (OP), and the chlorophenol pentachlorophenol (PCP)) on constructed wetlands (CWs) performance, including on organic matter, nutrients and contaminants removal efficiency, and on microbial community structure in the plant bed substrate. CWs were assembled at lab scale simulating a vertical flow configuration and irrigated along eight weeks with Ribeira de Joane (an urban stream) water not doped (control) or doped with a mixture of NP and OP or with PCP (at a 100 μg·L−1 concentration each). The presence of the phenolic contaminants did not interfere in the removal of organic matter or nutrients in CWs in the long term. Removals of NP and OP were >99%, whereas PCP removals varied between 87% and 98%, mainly due to biodegradation. Microbial richness, diversity and dominance in CWs substrate were generally not affected by phenolic compounds, with only PCP decreasing diversity. Microbial community structure, however, showed that there was an adaptation of the microbial community to the presence of each contaminant, with several specialist genera being enriched following exposure. The three more abundant specialist genera were Methylotenera and Methylophilus (methylophilaceae family) and Hyphomicrobium (hyphomicrobiaceae family) when the systems were exposed to a mixture of NP and OP. When exposed to PCP, the three more abundant genera were Denitromonas (Rhodocyclaceae family), Xenococcus_PCC_7305 (Xenococcaceae family) and Rhodocyclaceae_uncultured (Rhodocyclaceae family). To increase CWs efficiency in the elimination of phenolic compounds, namely PCP which was not totally removed, strategies to stimulate (namely biostimulation) or increase (namely bioaugmentation) the presence of these bacteria should be explore. This study clearly shows the potential of vertical flow CWs for the removal of phenolic compounds, a still little explored subject, contributing to promote the use of CWs as nature-based solutions to remediate water contaminated with different families of persistent and/or emergent contaminants.

Enhancing the Rice Seedlings Growth Promotion Abilities of Azoarcus sp. CIB by Heterologous Expression of ACC Deaminase to Improve Performance of Plants Exposed to Cadmium Stress.

Environmental pollutants can generate stress in plants causing increased ethylene production that leads to the inhibition of plant growth. Ethylene production by the stressed plant may be lowered by Plant Growth-Promoting Bacteria (PGPB) that metabolizes the immediate precursor of ethylene 1-aminocyclopropane-1-carboxylate (ACC). Thus, engineering PGPB with ACC deaminase activity can be a promising alternative to mitigate the harmful effects of pollutants and thus enhance plant production. Here we show that the aromatics-degrading and metal-resistant Azoarcus sp. CIB behaves as a PGP-bacterium when colonizing rice as an endophyte, showing a 30% increment in plant weight compared to non-inoculated plants. The cloning and expression of an acdS gene led to a recombinant strain, Azoarcus sp. CIB (pSEVA237acdS), possessing significant ACC deaminase activity (6716 nmol mg−1 h−1), constituting the first PGPB of the Rhodocyclaceae family equipped with this PGP trait. The recombinant CIB strain acquired the ability to protect inoculated rice plants from the stress induced by cadmium (Cd) exposure and to increase the Cd concentration in rice seedlings. The observed decrease of the levels of reactive oxygen species levels in rice roots confirms such a protective effect. The broad-host-range pSEVA237acdS plasmid paves the way to engineer PGPB with ACC deaminase activity to improve the growth of plants that might face stress conditions.

Adaptation of methane recovery, sludge characteristics and evolution of microbial community response to elevated nitrate under the methanogenic condition

Coupling of methanogenesis and denitrification with elevated nitrate under the methanogenic condition shows potential advantages for (N-) heterocyclic compounds removal and methane recovery simultaneously. However, methane production, sludge characteristics and microbial evolution response to elevated nitrate were seldom reported. Here, an experiment lasing about 300 days under the methanogenic condition with sequential influent nitrate addition as COD/NO3−-N ratio from 486.03 to 8.00 was carried out. More than 50% decline of removal rate of COD was observed when the COD/NO3−-N ratio varied from 19.97 to 8.00 (232 d). At last, the removal rate of COD recovered to around 94% at the COD/NO3−-N ratio of 8.00. About 90% of nitrate was removed through denitrification, and a lower content of methane (40–45%) and effluent acetate and propionate (<10 mg/L) were observed with elevated nitrate. Damage of the granule-shaped and multi-layer structure of the sludge with the lift of nitrate addition was studied. Results of high-throughput sequencing hinted that bacteria accounted for about 70% of total microbes and Proteobacteria, Bacteroidetes, Firmicute and Chloroflexi were the dominant phyla shared with sludge at different COD/NO3−-N ratio. Pseudomonas genus and Enterobacteriaceae family (Gammaproteobacteria class, Proteobacteria phylum), Brachymonas genus and Rhodocyclaceae family (Betaproteobacteria class, Proteobacteria phylum) might be vital for the denitrification process under the methanogenic condition. An increase of Smithella and Syntrophobacterale genus with elevated nitrate could be partly contributed to denitrification from the transformation of intermediates as propionate and butyrate. Methanosaeta and Methanobacterium genus were two major methanogens, and Methanosaeta seemed to be less adaptable for elevated nitrate than Methanobacterium. A suitable nitrate addition as influent COD/NO3−-N ratio ranged from 121.43 to 15.18 was recommended for the coupling of methanogenesis and denitrification. The results of this work would enhance the understanding of the effects and adaptation with elevated nitrate under the methanogenic condition and instruct engineering practice for low COD/NO3−-N wastewater treatment and methane recovery.

Induction of Microbial Oxidative Stress as a New Strategy to Enhance the Enzymatic Degradation of Organic Micropollutants in Synthetic Wastewater.

Organic micropollutants (OMPs) are pervasive anthropogenic contaminants of receiving waters where they can induce various adverse effects to aquatic life. Their ubiquitous environmental occurrence is primarily attributed to discharge from wastewater treatment plants due to incomplete removal by common biological wastewater treatment processes. Here, we assess a new strategy for promoting the degradation of six representative OMPs (i.e., sulfamethoxazole, carbamazepine, tylosin, atrazine, naproxen, and ibuprofen) by intentionally stimulating the production of microbial oxidoreductases to counter oxidative stress caused by oxygen perturbations. Mixed microbial cultures from a dairy farm wastewater were subjected to cyclic perturbations of dissolved oxygen (DO). A distance-based redundancy analysis was used to show that DO perturbations correlate with the abundance of Pseudomonadaceae and Rhodocyclaceae families, activities of peroxidases and cytochromes, and the degradation of OMPs. DO perturbation of 0.25 and 0.5 cycles/h led to most abundance of Pseudomonadaceae and Rhodocyclaceae families, showed higher activity of peroxidase and cytochrome, and gave largest removal of OMPs (removal of 92 ± 3% for sulfamethoxazole, 84 ± 3% for naproxen, 82 ± 3% for ibuprofen, 66 ± 2% for carbamazepine, 57 ± 15% for tylosin, and 88 ± 1% for atrazine).

Enhanced shortcut nitrogen removal and metagenomic analysis of functional microbial communities in a double sludge system treating ammonium-rich wastewater

ABSTRACTBiological nitrogen removal processes based on partial nitrification are promising for ammonium-rich wastewater treatment. In this study, a partial nitrification-denitrification double sludge system was applied to treat synthetic ammonium-rich wastewater. Metagenomic analysis of functional genes and metabolic pathways was conducted, also with the evaluation of system performance and nitrous oxide (N2O) emission. In the nitrifying sequencing batch reactor (SBRPN), the removal percentage of ammonium nitrogen reached to 99.98% with a high nitritation efficiency of 93.24%, and the N2O emission factor was 0.88%. In the denitrifying sequencing batch reactor (SBRDN), there was almost no nitrate nitrogen and nitrite nitrogen in the effluent, and the maximum N2O emission was 0.078 mg N/L. The dominant ammonia oxidizing bacteria was Nitrosomonas in SBRPN (13.6%), and the main potential denitrifiers in SBRDN were Thauera (14.6%), an uncultured genus in the Comamonadaceae family (4.0%), an uncultured genus in Rhodocyclaceae family (2.4%) and Comamonas (1.1%). Metagenomic analysis revealed that amo mainly distributed in Nitrosomonas eutropha (38.3%), Nitrosomonas europaea (27.1%), Nitrosomonas sp. GH22 (20.5%) and Nitrosomonas sp. TK794 (15.0%), and Bacteroidetes had the N2O reduction potential in SBRPN.

Microbiota of the Esophagus and Stomach in Patients with Gastroesophageal Reflux Disease and Healthy Volunteers

This paper is aimed at investigating the microbiota of the esophagus and stomach in patients with gastroesophageal reflux disease (GERD) and healthy volunteers.Materials and methods. The study included 15 patients suffering from GERD and 6 healthy volunteers. All subjects underwent sampling of esophageal and gastric contents. The study of the microbiota in the obtained samples was performed by sequencing the 16S gene of ribosomal RNA (rRNA).Results. The most common types of bacteria in the esophagus and stomach in patients with GERD and healthy volunteers are found to be Bacteroidetes, Firmicutes, Actinobacteria, Proteobacteria, Fusobacterium. By comparing the relative contents of the main types of bacteria in the esophageal mucus and gastric contents, a significant decrease in the proportion of Proteobacteria was observed in patients with GERD as compared to healthy volunteers. The decrease in the relative number of bacteria belonging to the Acetobacteraceae, Bacillaceae, Bdellovibrionaceae, Clostridiales Insertae Sedis XI, Fusobacteriaceae, Moraxellaceae, Pasteurellaceae and Rhodocyclaceae families was observed in the esophagus in patients with GERD as compared to healthy volunteers. A higher bacterial content of the Leptotrichiaceae and Veillonellaceae families was detected in the stomach of patients with GERD.Conclusions. The obtained results indicate differences in the intraluminal microbiota of the esophagus and stomach in patients with GERD and healthy volunteers. Further study should be carried out to study the effect of changes in bacterial composition on those in the esophagus and stomach.

Exploring microbial N2 O reduction: a continuous enrichment in nitrogen free medium.

N2 O is a potent greenhouse gas, but also a potent electron acceptor. In search of thermodynamically favourable - yet undescribed - metabolic pathways involving N2 O reduction, we set up a continuous microbial enrichment, inoculated with activated sludge, fed with N2 O as the sole electron acceptor and acetate as an electron donor. A nitrogen-free mineral medium was used with the intention of creating a selective pressure towards organisms that would use N2 O directly as source of nitrogen for cell synthesis. Instead, we obtained a culture dominated by microorganisms of the Rhodocyclaceae family growing by N2 O reduction to N2 coupled to N2 fixation. Biomass yields of this culture were 40% lower than those of a previously reported culture grown under comparable conditions but with an NH4+-amended medium, as expected from the extra energy expense of N2 fixation. Interestingly, we found no significant difference in yields whether N2 O or acetate was the growth-limiting substrate in the chemostat in contrast to the study with NH4+-amended medium, in which biomass yields were roughly 30% lower during acetate limiting conditions.

Candidatus Dactylopiibacterium carminicum, a Nitrogen-Fixing Symbiont of Dactylopius Cochineal Insects (Hemiptera: Coccoidea: Dactylopiidae).

The domesticated carmine cochineal Dactylopius coccus (scale insect) has commercial value and has been used for more than 500 years for natural red pigment production. Besides the domesticated cochineal, other wild Dactylopius species such as Dactylopius opuntiae are found in the Americas, all feeding on nutrient poor sap from native cacti. To compensate nutritional deficiencies, many insects harbor symbiotic bacteria which provide essential amino acids or vitamins to their hosts. Here, we characterized a symbiont from the carmine cochineal insects, Candidatus Dactylopiibacterium carminicum (betaproteobacterium, Rhodocyclaceae family) and found it in D. coccus and in D. opuntiae ovaries by fluorescent in situ hybridization, suggesting maternal inheritance. Bacterial genomes recovered from metagenomic data derived from whole insects or tissues both from D. coccus and from D. opuntiae were around 3.6 Mb in size. Phylogenomics showed that dactylopiibacteria constituted a closely related clade neighbor to nitrogen fixing bacteria from soil or from various plants including rice and other grass endophytes. Metabolic capabilities were inferred from genomic analyses, showing a complete operon for nitrogen fixation, biosynthesis of amino acids and vitamins and putative traits of anaerobic or microoxic metabolism as well as genes for plant interaction. Dactylopiibacterium nif gene expression and acetylene reduction activity detecting nitrogen fixation were evidenced in D. coccus hemolymph and ovaries, in congruence with the endosymbiont fluorescent in situ hybridization location. Dactylopiibacterium symbionts may compensate for the nitrogen deficiency in the cochineal diet. In addition, this symbiont may provide essential amino acids, recycle uric acid, and increase the cochineal life span.

Dynamics of the diversity and structure of the overall and nitrifying microbial community in activated sludge along gradient copper exposures.

Diversity and composition of the microbial community, especially the nitrifiers, are essential to the treatment efficiency of wastewater in activated sludge systems. Heavy metals commonly present in the wastewater influent such as Cu can alter the community structure of nitrifiers and lower their activity. However, the dynamics of microbial community along a gradient of metal exposure have largely been unexplored, partially due to the limitations in traditional molecular methods. This study explored the dynamics regarding the diversity and community structures of overall and nitrifying microbial communities in activated sludge under intermittent Cu gradient loadings using Illumina sequencing. We created a new local nitrifying bacterial database for sequence BLAST searches. High Cu loadings (>10.9mg/L) impoverished microbial diversity and altered the microbial community. Overall, Proteobacteria was the predominant phylum in the activated sludge system, in which Zoogloea, Thauera, and Dechloromonas (genera within the Rhodocyclaceae family of the Beta-proteobacteria class) were the dominant genera in the presence of Cu. The abundance of unclassified bacteria at the phylum level increased substantially with increasing Cu loadings. Nitrosomonas and Nitrospira were the predominant nitrifiers. The nitrifying bacterial community changed through increasing abundance and shifting to Cu-tolerant species to reduce the toxic effects of Cu. Our local nitrifying bacterial database helped to improve the resolution of bacterial identification. Our results provide insights into the dynamics of microbial community in response to various metal concentrations in activated sludge systems and improve our understanding regarding the effect of metals on wastewater treatment efficiency.

Uranium removal and microbial community in a H2-based membrane biofilm reactor

We evaluated a hydrogen-based membrane biofilm reactor (MBfR) for its capacity to reduce and remove hexavalent uranium [U(VI)] from water. After a startup period that allowed slow-growing U(VI) reducers to form biofilms, the MBfR successfully achieved and maintained 94–95% U(VI) removal over 8 months when the U surface loading was 6–11 e− mEq/m2-day. The MBfR biofilm was capable of self-recovery after a disturbance due to oxygen exposure. Nanocrystalline UO2 aggregates and amorphous U precipitates were associated with vegetative cells and apparently mature spores that accumulated in the biofilm matrix. Despite inoculation with a concentrated suspension of Desulfovibrio vulgaris, this bacterium was not present in the U(VI)-reducing biofilm. Instead, the most abundant group in the biofilm community contained U(VI) reducers in the Rhodocyclaceae family when U(VI) was the only electron acceptor. When sulfate was present, the community dramatically shifted to the Clostridiaceae family, which included spores that were potentially involved in U(VI) reduction.

The Impact of Environmental Heterogeneity and Life Stage on the Hindgut Microbiota of Holotrichia parallela Larvae (Coleoptera: Scarabaeidae)

Gut microbiota has diverse ecological and evolutionary effects on its hosts. However, the ways in which it responds to environmental heterogeneity and host physiology remain poorly understood. To this end, we surveyed intestinal microbiota of Holotrichia parallela larvae at different instars and from different geographic regions. Bacterial 16S rRNA gene clone libraries were constructed and clones were subsequently screened by DGGE and sequenced. Firmicutes and Proteobacteria were the major phyla, and bacteria belonging to Ruminococcaceae, Lachnospiraceae, Enterobacteriaceae, Desulfovibrionaceae and Rhodocyclaceae families were commonly found in all natural populations. However, bacterial diversity (Chao1 and Shannon indices) and community structure varied across host populations, and the observed variation can be explained by soil pH, organic carbon and total nitrogen, and the climate factors (e.g., mean annual temperature) of the locations where the populations were sampled. Furthermore, increases in the species richness and diversity of gut microbiota were observed during larval growth. Bacteroidetes comprised the dominant group in the first instar; however, Firmicutes composed the majority of the hindgut microbiota during the second and third instars. Our results suggest that the gut's bacterial community changes in response to environmental heterogeneity and host's physiology, possibly to meet the host's ecological needs or physiological demands.

Aerobic granular-type biomass development in a continuous stirred tank reactor

A continuous stirred tank reactor (CSTR) was operated in order to define the appropriate operational conditions to obtain aerobic biomass grown in the form of granules. A selection pressure of 10m/h of solids settling velocity was imposed while the hydraulic retention time (HRT) was gradually decreased from 6 to 1h to promote the wash-out of the suspended biomass. At HRT values of 6 and 3h, filamentous bacteria were dominant in the biomass. The formation of aerobic granular-type biomass was only achieved when the HRT was fixed at 1h. The physical properties of these granules were: sludge volumetric index of 127mL/g TSS, density of 11g VSS/Lgranule, settling velocity of 36–48m/h and average diameter of 6.8mm. Abundance of filamentous species was different at the tested HRTs. When the HRT was fixed at 6h filamentous shape bacteria appeared in large amounts, while at the HRT of 1h the bacterial shape was mainly bacillus and filamentous shape bacteria were absent. Microbial populations were mainly composed by members of Comamonadaceae and Rhodocyclaceae families within β-Proteobacteria, and members of the subclass γ-Proteobacteria.

Mechanism of uranium (VI) removal by two anaerobic bacterial communities

The mechanism of uranium (VI) removal by two anaerobic bacterial consortia, recovered from an uncontaminated site (consortium A) and other from an uranium mine (consortium U), was investigated. The highest efficiency of U (VI) removal by both consortia (97%) occurred at room temperature and at pH 7.2. Furthermore, it was found that U (VI) removal by consortium A occurred by enzymatic reduction and bioaccumulation, while the enzymatic process was the only mechanism involved in metal removal by consortium U. FTIR analysis suggested that after U (VI) reduction, U (IV) could be bound to carboxyl, phosphate and amide groups of bacterial cells. Phylogenetic analysis of 16S rRNA showed that community A was mainly composed by bacteria closely related to Sporotalea genus and Rhodocyclaceae family, while community U was mainly composed by bacteria related to Clostridium genus and Rhodocyclaceae family.

Evidence and Long-Term Feasibility of Enhanced Biological Phosphorus Removal in Oxidation-Ditch Type of Aerated-Anoxic Activated Sludge Systems

This study investigated the potential of four full-scale oxidation ditches to accomplish enhanced biological phosphorus removal (EBPR). Despite the fact that none of the tested oxidation ditches were designed to perform EBPR, mixed liquors from all four ditches showed good specific phosphorus release and uptake rates, a typical characteristic of a typical EBPR biomass. The specific phosphorus release rates ranged from 0.042- to 0.254-mg P/mg VSS-d and the specific phosphorus uptake rates ranged from 0.023- to 0.125-mg P/mg VSS-d for the tested full-scale plants. The EBPR potential of one of the full-scale plants (Central Davis Sewer District) was further studied by changing the aeration patterns in the ditch. The mixed liquor in this full-scale plant exhibited good phosphorus release and uptake trends and dissolved phosphorus, as low as 1.26 mg/L, could be accomplished in the final effluent of this plant as a result of this optimization. The long-term feasibility of the EBPR in this full-scale was tested by running a bench-scale EBPR reactor, in which the anaerobic phase was replaced with aerated-anaerobic phase to simulate the mixed liquor environment that exists in Central Davis mixed liquor and, in general, in all oxidation-ditch-type activated sludge configurations. The bench-scale reactor showed consistent EBPR activity in the absence of an anaerobic environment and more than 85% phosphorus removal could be maintained in the reactor for more than 400 days. The intrafloc microanaerobic/anoxic zones, which may be present in the mixed liquor, did not seem to add to the EBPR efficiency under aerated-anaerobic conditions. Cloning and sequencing using Rhodocyclus specific forward primer RHC439 showed the abundance of organisms most closely falling in Rhodocyclaceae family but not related to CandidatusAccumulibacter phosphatis. Simultaneous 4′-6–Diamidino-2–phenylindole (DAPI) staining and fluorescent in situ hybridization (FISH) using RHC439 probe clearly demonstrated the participation of polyphosphate accumulating organism (PAOs) targeted by RHC439 (i.e., in Rhodocyclaceae family). Microautoradiography assisted FISH using RHC439 further confirmed the participation of PAOs in Rhodocyclaceae family.

Effect of uranium (VI) on two sulphate-reducing bacteria cultures from a uranium mine site

This work was conducted to assess the impact of uranium (VI) on sulphate-reducing bacteria (SRB) communities obtained from environmental samples collected on the Portuguese uranium mining area of Urgeiriça. Culture U was obtained from a sediment, while culture W was obtained from sludge from the wetland of that mine. Temperature gradient gel electrophoresis (TGGE) was used to monitor community changes under uranium stress conditions. TGGE profiles of dsrB gene fragment demonstrated that the initial cultures were composed of SRB species affiliated with Desulfovibrio desulfuricans, Desulfovibrio vulgaris and Desulfomicrobium spp. (sample U), and by species related to D. desulfuricans (sample W). A drastic change in SRB communities was observed as a result of uranium (VI) exposure. Surprisingly, SRB were not detected in the uranium removal communities. Such findings emphasize the need of monitoring the dominant populations during bio-removal studies. TGGE and phylogenetic analysis of the 16S rRNA gene fragment revealed that the uranium removal consortia are composed by strains affiliated to Clostridium genus, Caulobacteraceae and Rhodocyclaceae families. Therefore, these communities can be attractive candidates for environmental biotechnological applications associated to uranium removal.

Anaerobic bio-removal of uranium (VI) and chromium (VI): Comparison of microbial community structure

Several microbial communities, obtained from uranium contaminated and non-contaminated samples, were investigated for their ability to remove uranium (VI) and the cultures capable for this removal were further assessed on their efficiency for chromium (VI) removal. The highest efficiency for removal of both metals was observed on a consortium from a non-contaminated soil collected in Monchique thermal place, which was capable to remove 91% of 22 mg L −1 U(VI) and 99% of 13 mg L −1 Cr(VI). This study revealed that uranium (VI) removing communities have also ability to remove chromium (VI), but when uranium (VI) was replaced by chromium (VI) several differences in the structure of all bacterial communities were observed. TGGE and phylogenetic analysis of 16S rRNA gene showed that the uranium (VI) removing bacterial consortia are mainly composed by members of Rhodocyclaceae family and Clostridium genus. On the other hand, bacteria from Enterobacteriaceae family were detected in the community with ability for chromium (VI) removal. The existence of members of Enterobacteriaceae and Rhodocyclaceae families never reported as chromium or uranium removing bacteria, respectively, is also a relevant finding, encouraging the exploitation of microorganisms with new abilities that can be useful for bioremediation.