Repeated replacement of an intrabacterial symbiont in the tripartite nested mealybug symbiosis.

Stable endosymbiosis of a bacterium into a host cell promotes cellular and genomic complexity. The mealybug Planococcus citri has two bacterial endosymbionts with an unusual nested arrangement: the γ-proteobacterium Moranella endobia lives in the cytoplasm of the ß-proteobacterium Tremblaya princeps These two bacteria, along with genes horizontally transferred from other bacteria to the P. citri genome, encode gene sets that form an interdependent metabolic patchwork. Here, we test the stability of this three-way symbiosis by sequencing host and symbiont genomes for five diverse mealybug species and find marked fluidity over evolutionary time. Although Tremblaya is the result of a single infection in the ancestor of mealybugs, the γ-proteobacterial symbionts result from multiple replacements of inferred different ages from related but distinct bacterial lineages. Our data show that symbiont replacement can happen even in the most intricate symbiotic arrangements and that preexisting horizontally transferred genes can remain stable on genomes in the face of extensive symbiont turnover.

Insights into selenate removal mechanism of hydrogen-based membrane biofilm reactor for nitrate-polluted groundwater treatment based on anaerobic biofilm analysis.

The selenate removal mechanism of hydrogen-based membrane biofilm reactor (MBfR) for nitrate-polluted groundwater treatment was studied based on anaerobic biofilm analysis. A laboratory-scale MBfR was operated for over 60 days with electron balance, structural analysis, and bacterial community identification. Results showed that anaerobic biofilm had an excellent removal of both selenate (95%) and nitrate (100%). Reduction of Selenate → Selenite → Se0 with hydrogen was the main pathway of anaerobic biofilm for selenate removal with amorphous Se0 precipitate accumulating in the biofilm. The element selenium was observed to be evenly distributed along the cross-sectional thin biofilm. A part of selenate (3%) was also reduced into methyl-selenide by heterotrophic bacteria. Additionally, Hydrogenophaga bacteria of ß-Proteobacteria, capable of both nitrate and selenate removal, worked as the dominant species (over 85%) in the biofilm and contributed to the stable removal of both nitrate and selenate. With the selenate input, bacteria with a capacity for both selenate and nitrate removal were also developed in the anaerobic biofilm community.

Epibiont growth on filamentous bacteria found in activated sludge: a morphological approach.

Occurrence of epibiont attachment on filamentous bacteria is a common phenomenon in activated sludge. In this study, an attempt has been made to elucidate the intrinsic nature of the attachment between the epibionts and filamentous bacteria based on microscopic observations. Characterization of the epiflora based on fluorescence in situ hybridization using group level probes revealed that the epibionts colonizing these filamentous bacteria largely belongs to the class Alphaproteobacteria, followed by Beta and Gammaproteobacteria. The ultrastructural examination using transmission electron microscopy pointed to the existence of a possible cell-to-cell interaction between epibionts and the selected filaments. Common bacterial appendages such as pili and fimbria were absent at the interface and further noted was the presence of cell membrane extensions on epibiont bacteria protruding towards the targeted filamentous cell. Fibrillar structures resembling amyloid-like proteins were observed within the filament cells targeted by the epibionts. An interaction was apparent between amyloid such as proteins and epibionts with regards to the direction of fibrillar structures and the distance of approaching epibiont bacteria. Due to the lack of visual evidence in support of penetration, the role of these amyloid-like fibrils as potential attachment sites for the epibionts was taken into consideration, and required further validation using conformational antibodies.

The reduced genome of Candidatus Kinetoplastibacterium sorsogonicusi, the endosymbiont of Kentomonas sorsogonicus (Trypanosomatidae): loss of the haem-synthesis pathway.

Trypanosomatids of the genera Angomonas and Strigomonas (subfamily Strigomonadinae) have long been known to contain intracellular beta-proteobacteria, which provide them with many important nutrients such as haem, essential amino acids and vitamins. Recently, Kentomonas sorsogonicus, a divergent member of Strigomonadinae, has been described. Herein, we characterize the genome of its endosymbiont, Candidatus Kinetoplastibacterium sorsogonicusi. This genome is completely syntenic with those of other known Ca. Kinetoplastibacterium spp., but more reduced in size (~742 kb, compared with 810-833 kb, respectively). Gene losses are not concentrated in any hot-spots but are instead distributed throughout the genome. The most conspicuous loss is that of the haem-synthesis pathway. For long, removing haemin from the culture medium has been a standard procedure in cultivating trypanosomatids isolated from insects; continued growth was considered as an evidence of endosymbiont presence. However, we demonstrate that, despite bearing the endosymbiont, K. sorsogonicus cannot grow in culture without haem. Thus, the traditional test cannot be taken as a reliable criterion for the absence or presence of endosymbionts in trypanosomatid flagellates. It remains unclear why the ability to synthesize such an essential compound was lost in Ca. K. sorsogonicusi, whereas all other known bacterial endosymbionts of trypanosomatids retain them.

Microbial N Transformations and N2O Emission after Simulated Grassland Cultivation: Effects of the Nitrification Inhibitor 3,4-Dimethylpyrazole Phosphate (DMPP).

Grassland cultivation can mobilize large pools of N in the soil, with the potential for N leaching and N2O emissions. Spraying with the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) before cultivation was simulated by use of soil columns in which the residue distribution corresponded to plowing or rotovation to study the effects of soil-residue contact on N transformations. DMPP was sprayed on aboveground parts of ryegrass and white clover plants before incorporation. During a 42-day incubation, soil mineral N dynamics, potential ammonia oxidation (PAO), denitrifying enzyme activity (DEA), nitrifier and denitrifier populations, and N2O emissions were investigated. The soil NO3- pool was enriched with 15N to trace sources of N2O. Ammonium was rapidly released from decomposing residues, and PAO was stimulated in soil near residues. DMPP effectively reduced NH4+ transformation irrespective of residue distribution. Ammonia-oxidizing archaea (AOA) and bacteria (AOB) were both present, but only the AOB amoA transcript abundance correlated with PAO. DMPP inhibited the transcription of AOB amoA genes. Denitrifier genes and transcripts (nirK, nirS, and clades I and II of nosZ) were recovered, and a correlation was found between nirS mRNA and DEA. DMPP showed no adverse effects on the abundance or activity of denitrifiers. The 15N enrichment of N2O showed that denitrification was responsible for 80 to 90% of emissions. With support from a control experiment without NO3- amendment, it was concluded that DMPP will generally reduce the potential for leaching of residue-derived N, whereas the effect of DMPP on N2O emissions will be significant only when soil NO3- availability is limiting. IMPORTANCE: Residue incorporation following grassland cultivation can lead to mobilization of large pools of N and potentially to significant N losses via leaching and N2O emissions. This study proposed a mitigation strategy of applying 3,4-dimethylpyrazole phosphate (DMPP) prior to grassland cultivation and investigated its efficacy in a laboratory incubation study. DMPP inhibited the growth and activity of ammonia-oxidizing bacteria but had no adverse effects on ammonia-oxidizing archaea and denitrifiers. DMPP can effectively reduce the potential for leaching of NO3- derived from residue decomposition, while the effect on reducing N2O emissions will be significant only when soil NO3- availability is limiting. Our findings provide insight into how DMPP affects soil nitrifier and denitrifier populations and have direct implications for improving N use efficiency and reducing environmental impacts during grassland cultivation.

Mechanism of electron transport during thiosulfate oxidation in an obligately mixotrophic bacterium Thiomonas bhubaneswarensis strain S10 (DSM 18181T).

This study describes the thiosulfate-supported respiratory electron transport activity of Thiomonas bhubaneswarensis strain S10 (DSM 18181T). Whole-genome sequence analysis revealed the presence of complete sox (sulfur oxidation) gene cluster (soxCDYZAXB) including the sulfur oxygenase reductase (SOR), sulfide quinone reductase (SQR), sulfide dehydrogenase (flavocytochrome c (fcc)), thiosulfate dehydrogenase (Tsd), sulfite dehydrogenase (SorAB), and intracellular sulfur oxidation protein (DsrE/DsrF). In addition, genes encoding respiratory electron transport chain components viz. complex I (NADH dehydrogenase), complex II (succinate dehydrogenase), complex III (ubiquinone-cytochrome c reductase), and various types of terminal oxidases (cytochrome c and quinol oxidase) were identified in the genome. Using site-specific electron donors and inhibitors and by analyzing the cytochrome spectra, we identified the shortest thiosulfate-dependent electron transport chain in T. bhubaneswarensis DSM 18181T. Our results showed that thiosulfate supports the electron transport activity in a bifurcated manner, donating electrons to quinol (bd) and cytochrome c (Caa 3 ) oxidase; these two sites (quinol oxidase and cytochrome c oxidase) also showed differences in their phosphate esterification potential (oxidative phosphorylation efficiency (P/O)). Further, it was evidenced that the substrate-level phosphorylation is the major contributor to the total energy budget in this bacterium.

Rethinking growth and decay kinetics in activated sludge - towards a new adaptive kinetics approach.

Growth kinetics in activated sludge modelling (ASM) are typically assumed to be the result of intrinsic growth and decay properties and thus process parameters are deemed to be constant. The activity change in a microbial population is expressed in terms of variance of the active biomass fraction and not actual shifts in bacterial cellular activities. This approach is limited, in that it does not recognise the reality that active biomass is highly physiologically adaptive. Here, a strong correlation between maximum specific growth rate (µmax) and decay rate (be) of ordinary heterotrophic organisms was revealed in both low solids retention times (SRT) and high SRT activated sludge systems. This relationship is indicative of physiological adaptation either for growth (high µmax and be) or survival optimization (low µmax and be). Further, the nitrifier decay process was investigated using molecular techniques to measure decay rates of ammonia oxidizing bacteria and nitrite oxidizing bacteria over a range of temperatures. This approach revealed decay rates 10-12% lower than values previously accepted and used in ASM. These findings highlight potential benefits of incorporating physiological adaptation of heterotrophic and nitrifying populations in future ASM.

Predicting N2O emissions from nitrifying and denitrifying biofilms: a modeling study.

Wastewater treatment plants can be significant sources of nitrous oxide (N2O), a potent greenhouse gas. While our understanding of N2O emissions from suspended-growth processes has advanced significantly, less is known about emissions from biofilm processes. Biofilms may behave differently due to their substrate gradients and microbial stratification. In this study, we used mathematical modeling to explore the mechanisms of N2O emissions from nitrifying and denitrifying biofilms. Our ammonia-oxidizing bacteria biofilm model suggests that N2O emissions from biofilm can be significantly greater than from suspended-growth systems. The driving factor is the diffusion of hydroxylamine, a nitrification intermediate, from the aerobic to the anoxic regions of the biofilm. The presence of nitrite-oxidizing bacteria further increased emissions. For denitrifying biofilms, our results suggest that emissions are generally greater than for suspended-growth systems. However, the magnitude of the difference depends on the bulk dissolved oxygen, chemical oxygen demand, and nitrate concentrations, as well as the biofilm thickness. Overall, the accumulation and diffusion of key intermediates, i.e. hydroxylamine and nitrite, distinguish biofilms from suspended-growth systems. Our research suggests that the mechanisms of N2O emissions from biofilms are much more complex than suspended-growth systems, and that emissions may be higher in many cases.

Comparison of different two-pathway models for describing the combined effect of DO and nitrite on the nitrous oxide production by ammonia-oxidizing bacteria.

The aim of this work is to compare the capability of two recently proposed two-pathway models for predicting nitrous oxide (N2O) production by ammonia-oxidizing bacteria (AOB) for varying ranges of dissolved oxygen (DO) and nitrite. The first model includes the electron carriers whereas the second model is based on direct coupling of electron donors and acceptors. Simulations are confronted to extensive sets of experiments (43 batches) from different studies with three different microbial systems. Despite their different mathematical structures, both models could well and similarly describe the combined effect of DO and nitrite on N2O production rate and emission factor. The model-predicted contributions for nitrifier denitrification pathway and hydroxylamine pathway also matched well with the available isotopic measurements. Based on sensitivity analysis, calibration procedures are described and discussed for facilitating the future use of those models.

Betaproteobacteria Limnohabitans strains increase fecundity in the crustacean Daphnia magna: symbiotic relationship between major bacterioplankton and zooplankton in freshwater ecosystem.

How symbioses between bacteria and aquatic animals influence food webs in freshwater ecosystems is a fundamental question in ecology. We investigated symbiosis between a crustacean zooplankton Daphnia magna and its dominant bacterial symbiont Limnohabitans, an abundant and globally distributed freshwater Betaproteobacteria. Aposymbiotic juvenile Daphnia were prepared and exposed to any of four Limnohabitans sp. - Limnohabitans strains DM1, 2KL-3, 2KL-7 and Limnohabitans planktonicus strain II-D5, all previously found in D. magna digestive tract or culture. Re-infected Daphnia were cultured until they produced the first clutch of juveniles. Limnohabitans strain DM1 and L. planktonicus strain II-D5 successfully re-infected Daphnia through single exposure at the first instar juvenile stage. In contrast to aposymbiotic Daphnia that produced non-viable juveniles, re-infected Daphnia produced viable juveniles and increased fecundity to levels of that of symbiotic Daphnia. Re-infected Daphnia did not increase their number of eggs nor growth rates. Limnohabitans strains 2KL-7 and 2KL-3 could not recover fecundity even in multiple exposures during culture. This study shows the functional evidence demonstrating that a single bacterium Limnohabitans regulates fecundity of the consumer Daphnia through symbiosis. Our results indicated that symbiotic relationship between major bacterioplankton and zooplankton is important for maintaining the population of zooplankton in freshwater ecosystems.

Nitrous Oxide Reduction Kinetics Distinguish Bacteria Harboring Clade I NosZ from Those Harboring Clade II NosZ.

UNLABELLED: Bacteria capable of reduction of nitrous oxide (N2O) to N2 separate into clade I and clade II organisms on the basis of nos operon structures and nosZ sequence features. To explore the possible ecological consequences of distinct nos clusters, the growth of bacterial isolates with either clade I (Pseudomonas stutzeri strain DCP-Ps1, Shewanella loihica strain PV-4) or clade II (Dechloromonas aromatica strain RCB, Anaeromyxobacter dehalogenans strain 2CP-C) nosZ with N2O was examined. Growth curves did not reveal trends distinguishing the clade I and clade II organisms tested; however, the growth yields of clade II organisms exceeded those of clade I organisms by 1.5- to 1.8-fold. Further, whole-cell half-saturation constants (Kss) for N2O distinguished clade I from clade II organisms. The apparent Ks values of 0.324 ± 0.078 µM for D. aromatica and 1.34 ± 0.35 µM for A. dehalogenans were significantly lower than the values measured for P. stutzeri (35.5 ± 9.3 µM) and S. loihica (7.07 ± 1.13 µM). Genome sequencing demonstrated that Dechloromonas denitrificans possessed a clade II nosZ gene, and a measured Ks of 1.01 ± 0.18 µM for N2O was consistent with the values determined for the other clade II organisms tested. These observations provide a plausible mechanistic basis for why the relative activity of bacteria with clade I nos operons compared to that of bacteria with clade II nos operons may control N2O emissions and determine a soil's N2O sink capacity. IMPORTANCE: Anthropogenic activities, in particular fertilizer application for agricultural production, increase N2O emissions to the atmosphere. N2O is a strong greenhouse gas with ozone destruction potential, and there is concern that nitrogen may become the major driver of climate change. Microbial N2O reductase (NosZ) catalyzes N2O reduction to environmentally benign dinitrogen gas and represents the major N2O sink process. The observation that bacterial groups with clade I nosZ versus those with clade II nosZ exhibit distinct affinities to N2O has implications for N2O flux models, and these distinct characteristics may provide opportunities to curb N2O emissions from relevant soil ecosystems.

Seasonal growth potential of rare lake water bacteria suggest their disproportional contribution to carbon fluxes.

We studied the seasonal growth potential of opportunistic bacterial populations in Lake Zurich (Switzerland) by a series of grazer-free dilution culture assays. Pronounced shifts in the composition of the bacterial assemblages were observed within one doubling of total cell numbers, from initially abundant Actinobacteria to other fast-growing microbial lineages. Small populations with growth potentials far above community average were detected throughout the year with striking seasonal differences in their respective taxonomic affiliations. Members of Cytophaga-Flavobacteria (CF) were disproportionally proliferating only during phytoplankton blooms in spring and summer, while Beta- and Gammaproteobacteria showed superior growth at all other occasions. Growth rates of Alphaproteobacteria and esp. Sphingomonadaceae were significantly correlated to water temperatures and were far above community average in summer. Within the genus Flavobacterium, two species-like populations showed a tendency for fast growth in most experiments, while four others were exclusively proliferating either during a spring or during a summer phytoplankton bloom. Their high growth potentials but low in situ abundances hint at a tight control by bacterivorous grazers and at a consequently accelerated carbon flux to higher trophic levels.

Assessment of microbial communities in mung bean (Vigna radiata) rhizosphere upon exposure to phytotoxic levels of Copper.

Pollution of agricultural soils by Cu is of concern as it could bring about alterations in microbial communities, ultimately eliminating certain plant beneficial bacteria thus disturbing soil fertility and plant growth. To understand the response of rhizobacterial communities upon Cu perturbation, mung bean (Vigna radiata) plants were grown in agricultural soil amended with CuSO4 (0-1000 mg kg(-1) ) under laboratory conditions. Culture-independent and -dependent Denaturing Gradient Gel Electrophoresis (CI-DGGE and CD-DGGE) fingerprinting techniques were employed to monitor rhizobacterial community shifts upon Cu amendment. In group specific PCR-DGGE, a negative impact was seen on α-Proteobacteria followed by ß-Proteobacteria resulting in a concomitant decrease in diversity indices with increased Cu concentration. No significant changes were observed in Firmicutes and Actinomycetes populations. In CD-DGGE rhizobacterial community shift was observed above 500 mg kg(-1) (CuSO4 ), however certain bands were predominantly present in all treatments. Plants showed toxic effects by reduction in growth and elevated Cu accumulation, with root system being affected prominently. From this study it is evident that above 250 mg kg(-1) , rhizobacterial communities are adversely affected. α-Proteobacteria was found to be a sensitive bio-indicator for Cu toxicity and is of particular significance since this group includes majority of plant growth promoting rhizobacteria.

Coproporphyrin III excretion identifies the anaerobic coproporphyrinogen III oxidase HemN as a copper target in the Cu⁺-ATPase mutant copA⁻ of Rubrivivax gelatinosus.

Two genes encoding structurally similar Copper P1B -type ATPases can be identified in several genomes. Notwithstanding the high sequence and structural similarities these ATPases held, it has been suggested that they fulfil distinct physiological roles. In deed, we have shown that the Cu(+) -ATPase CtpA is required only for the activity of cuproproteins in the purple bacterium Rubrivivax gelatinosus; herein, we show that CopA is not directly required for cytochrome c oxidase but is vital for copper tolerance. Interestingly, excess copper in the copA(-) mutant resulted in a substantial decrease of the cytochrome c oxidase and the photosystem under microaerobic and anaerobic conditions together with the extrusion of coproporphyrin III. The data indicated that copper targeted the tetrapyrrole biosynthesis pathway at the level of the coproporphyrinogen III oxidase HemN and thereby affects the oxidase and the photosystem. This is the first in vivo demonstration that copper, like oxygen, affects tetrapyrrole biosynthesis presumably at the level of the SAM and [4Fe-4S] containing HemN enzyme. In light of these results and similar findings in Escherichia coli, the potential role of copper ions in the evolution of [4Fe-4S] enzymes and the Cu(+) -ATPases is discussed.

Uncovering a microbial enigma: isolation and characterization of the streamer-generating, iron-oxidizing, acidophilic bacterium "Ferrovum myxofaciens".

A betaproteobacterium, shown by molecular techniques to have widespread global distribution in extremely acidic (pH 2 to 4) ferruginous mine waters and also to be a major component of "acid streamer" growths in mine-impacted water bodies, has proven to be recalcitrant to enrichment and isolation. A modified "overlay" solid medium was devised and used to isolate this bacterium from a number of mine water samples. The physiological and phylogenetic characteristics of a pure culture of an isolate from an abandoned copper mine ("Ferrovum myxofaciens" strain P3G) have been elucidated. "F. myxofaciens" is an extremely acidophilic, psychrotolerant obligate autotroph that appears to use only ferrous iron as an electron donor and oxygen as an electron acceptor. It appears to use the Calvin-Benson-Bassham pathway to fix CO2 and is diazotrophic. It also produces copious amounts of extracellular polymeric materials that cause cells to attach to each other (and to form small streamer-like growth in vitro) and to different solid surfaces. "F. myxofaciens" can catalyze the oxidative dissolution of pyrite and, like many other acidophiles, is tolerant of many (cationic) transition metals. "F. myxofaciens" and related clone sequences form a monophyletic group within the Betaproteobacteria distantly related to classified orders, with genera of the family Nitrosomonadaceae (lithoautotrophic, ammonium-oxidizing neutrophiles) as the closest relatives. On the basis of the phylogenetic and phenotypic differences of "F. myxofaciens" and other Betaproteobacteria, a new family, "Ferrovaceae," and order, "Ferrovales," within the class Betaproteobacteria are proposed. "F. myxofaciens" is the first extreme acidophile to be described in the class Betaproteobacteria.

Potential role of nitrite for abiotic Fe(II) oxidation and cell encrustation during nitrate reduction by denitrifying bacteria.

Microorganisms have been observed to oxidize Fe(II) at neutral pH under anoxic and microoxic conditions. While most of the mixotrophic nitrate-reducing Fe(II)-oxidizing bacteria become encrusted with Fe(III)-rich minerals, photoautotrophic and microaerophilic Fe(II) oxidizers avoid cell encrustation. The Fe(II) oxidation mechanisms and the reasons for encrustation remain largely unresolved. Here we used cultivation-based methods and electron microscopy to compare two previously described nitrate-reducing Fe(II) oxidizers ( Acidovorax sp. strain BoFeN1 and Pseudogulbenkiania sp. strain 2002) and two heterotrophic nitrate reducers (Paracoccus denitrificans ATCC 19367 and P. denitrificans Pd 1222). All four strains oxidized ∼8 mM Fe(II) within 5 days in the presence of 5 mM acetate and accumulated nitrite (maximum concentrations of 0.8 to 1.0 mM) in the culture media. Iron(III) minerals, mainly goethite, formed and precipitated extracellularly in close proximity to the cell surface. Interestingly, mineral formation was also observed within the periplasm and cytoplasm; intracellular mineralization is expected to be physiologically disadvantageous, yet acetate consumption continued to be observed even at an advanced stage of Fe(II) oxidation. Extracellular polymeric substances (EPS) were detected by lectin staining with fluorescence microscopy, particularly in the presence of Fe(II), suggesting that EPS production is a response to Fe(II) toxicity or a strategy to decrease encrustation. Based on the data presented here, we propose a nitrite-driven, indirect mechanism of cell encrustation whereby nitrite forms during heterotrophic denitrification and abiotically oxidizes Fe(II). This work adds to the known assemblage of Fe(II)-oxidizing bacteria in nature and complicates our ability to delineate microbial Fe(II) oxidation in ancient microbes preserved as fossils in the geological record.

Population dynamics and growth rates of endosymbionts during Diaphorina citri (Hemiptera, Liviidae) ontogeny.

The infection density of symbionts is among the major parameters to understand their biological effects in host-endosymbionts interactions. Diaphorina citri harbors two bacteriome-associated bacterial endosymbionts (Candidatus Carsonella ruddii and Candidatus Profftella armatura), besides the intracellular reproductive parasite Wolbachia. In this study, the density dynamics of the three endosymbionts associated with the psyllid D. citri was investigated by real-time quantitative PCR (qPCR) at different developmental stages. Bacterial density was estimated by assessing the copy number of the 16S rRNA gene for Carsonella and Profftella, and of the ftsZ gene for Wolbachia. Analysis revealed a continuous growth of the symbionts during host development. Symbiont growth and rate curves were estimated by the Gompertz equation, which indicated a negative correlation between the degree of symbiont-host specialization and the time to achieve the maximum growth rate (t*). Carsonella densities were significantly lower than those of Profftella at all host developmental stages analyzed, even though they both displayed a similar trend. The growth rates of Wolbachia were similar to those of Carsonella, but Wolbachia was not as abundant. Adult males displayed higher symbiont densities than females. However, females showed a much more pronounced increase in symbiont density as they aged if compared to males, regardless of the incorporation of symbionts into female oocytes and egg laying. The increased density of endosymbionts in aged adults differs from the usual decrease observed during host aging in other insect-symbiont systems.

Survival of prokaryotes in a polluted waste dump during remediation by alkaline hydrolysis.

A combination of culture-dependent and culture-independent techniques was used to characterize bacterial and archaeal communities in a highly polluted waste dump and to assess the effect of remediation by alkaline hydrolysis on these communities. This waste dump (Breakwater 42), located in Denmark, contains approximately 100 different toxic compounds including large amounts of organophosphorous pesticides such as parathions. The alkaline hydrolysis (12 months at pH >12) decimated bacterial and archaeal abundances, as estimated by 16S rRNA gene-based qPCR, from 2.1 × 10(4) and 2.9 × 10(3) gene copies per gram wet soil respectively to below the detection limit of the qPCR assay. Clone libraries constructed from PCR-amplified 16S rRNA gene fragments showed a significant reduction in bacterial diversity as a result of the alkaline hydrolysis, with preferential survival of Betaproteobacteria, which increased in relative abundance from 0 to 48 %. Many of the bacterial clone sequences and the 27 isolates were related to known xenobiotic degraders. An archaeal clone library from a non-hydrolyzed sample showed the presence of three main clusters, two representing methanogens and one representing marine aerobic ammonia oxidizers. Isolation of alkalitolerant bacterial pure cultures from the hydrolyzed soil confirmed that although alkaline hydrolysis severely reduces microbial community diversity and size certain bacteria survive a prolonged alkaline hydrolysis process. Some of the isolates from the hydrolyzed soil were capable of growing at high pH (pH 10.0) in synthetic media indicating that they could become active in in situ biodegradation upon hydrolysis.

Strengthening the growth of Rubrivivax gelatinosus in sewage purification through ferric ion regulated photophosphorylation and respiration.

Rubrivivax gelatinosus has the potential of biomass resource recycling combined with sewage purification. However, low biomass production and yield restricts the potential for sewage purification. Thus, this research investigated the improvement of biomass production and yield and organics reduction by Fe(3+) in R. gelatinosus wastewater treatment. Results showed that 10-30 mg/L Fe(3+) improved biomass yield in wastewater to a level found in culture medium. With optimal dosage (20 mg/L), biomass production reached 4,300 mg/L, which was 1.67 times that of the control group. Biomass yield was improved by 43.3%. Chemical oxygen demand (COD) removal reached above 91%. Hydraulic retention time was shortened by 25%. Mechanism analysis indicated that Fe(3+) enhanced the succinate and NADH dehydrogenase activities and, bacteriochlorophyll content in three energy metabolism pathways. These effects then enhanced adenosine triphosphate (ATP) production, which led to more biomass accumulation and COD removal. With 20 mg/L Fe(2+) dosage, succinate and NADH dehydrogenase, coproporphyrinogen III oxidase activities, bacteriochlorophyll content and ATP production were improved, respectively, by 48.4, 50.8, 50, 67 and 56% compared to those of the control group.

Photo-induced electron transfer in intact cells of Rubrivivax gelatinosus mutants deleted in the RC-bound tetraheme cytochrome: insight into evolution of photosynthetic electron transport.

Deletion of two of the major electron carriers, the reaction center-bound tetrahemic cytochrome and the HiPIP, involved in the light-induced cyclic electron transfer pathway of the purple photosynthetic bacterium, Rubrivivax gelatinosus, significantly impairs its anaerobic photosynthetic growth. Analysis on the light-induced absorption changes of the intact cells of the mutants shows, however, a relatively efficient photo-induced cyclic electron transfer. For the single mutant lacking the reaction center-bound cytochrome, we present evidence that the electron carrier connecting the reaction center and the cytochrome bc(1) complex is the High Potential Iron-sulfur Protein. In the double mutant lacking both the reaction center-bound cytochrome and the High Potential Iron-sulfur Protein, this connection is achieved by the high potential cytochrome c(8). Under anaerobic conditions, the halftime of re-reduction of the photo-oxidized primary donor by these electron donors is 3 to 4 times faster than the back reaction between P(+) and the reduced primary quinone acceptor. This explains the photosynthetic growth of these two mutants. The results are discussed in terms of evolution of the type II RCs and their secondary electron donors.