Biomedical subjects

R Moletta

Publications and source records attributed to R Moletta.

At least 37 records · Page 2Linked to original sources

PubMed2001

Evaluation of the denitrifying microbiota of anoxic reactors.

Removal of inorganic nitrogen compounds from wastewaters can be accomplished by a combination of the biological processes of nitrification and denitrification. The information on the microbiota present in denitrifying reactors is still scarce. In the present work the evaluation of the denitrifying microbiota of different reactor sludges was performed by specific activity measurements and MPN count of denitrifiers. We also present the isolation and physiological and phylogenetic characterisation of denitrifying bacteria from the anoxic reactor of a combined system treating landfill leachate. Specific denitrifying activity measurements were faster to perform and more reliable than MPN enumerations. 16S rDNA characterisation of the isolates showed that they belonged to the genera Thauera, Acidovorax and Alcaligenes and were closely related to microorganisms retrieved from ecosystems rich in recalcitrant compounds. Two of the isolates could grow on aromatic compounds as sole carbon source.

Journal Article↗

PubMed2001

New perspectives in anaerobic digestion.

The IWA specialised group on anaerobic digestion (AD) is one of the oldest working groups of the former IAWQ organisation. Despite the fact that anaerobic technology dates back more than 100 years, the technology is still under development, adapting novel treatment systems to the modern requirements. In fact, most advances were achieved during the last three decades, when high-rate reactor systems were developed and a profound insight was obtained in the microbiology of the anaerobic communities. This insight led to a better understanding of anaerobic treatment and, subsequently, to a broader application potential. The present "state-of-the-art" paper, which has been written by members of the AD management committee, reflects the latest achievements and sets future lines for further development.

Bacteria, Aerobic↗

PubMed2001

A molecular method to study population and activity dynamics in anaerobic digestors.

The applicability of a new molecular fingerprinting method (Single Strand Conformation Polymorphism) to study the microbial populations of anaerobic digestors was investigated. After extraction of total nucleic acids, the 16S rDNA and 16S rRNA molecules were amplified and the amplicons were separated by SSCP electrophoresis. Characteristic and complex peak patterns were obtained, where each peak could be correlated with the 16S rDNA sequence of one micro-organism. The rDNA peak patterns should consist of the most abundant sequences and thus would reflect the diversity of prominent species of different digestors. Ribosomal DNA patterns were compared to rRNA patterns and revealed the bacteria that were the most active metabolically. The SSCP method also revealed dynamic changes in the presence and activity of populations, following perturbations such as an acidic shock which caused an increase in activity of two species. After cloning the 16S rDNA, the species corresponding to the peaks of interest, such as the archaeal species, could be identified by screening the clones according to their SSCP patterns and sequencing the 16S rDNA.

Bacteria, Anaerobic↗

PubMed2001

Monitoring of the microbial community of a sequencing batch reactor bioaugmented to improve its phosphorus removal capabilities.

The acclimatisation of an activated sludge to enhanced biological phosphorus removal conditions was followed after and without bioaugmentation with a low amount of phosphorus-accumulating sludge. Phosphorus removal yields were monitored by conventional analytical methods and microbial communities evolutions were followed by a finger printing molecular technique (PCR-SSCP). While the benefit of the bioaugmentation seems real at the level of the reactor parameters, bioaugmentation speeded up the installation of good and stable phosphorus removal yield, the establishment of the inoculated microbial community in the bioaugmented reactor is still unclear. Both the bioaugmented and the control microbial communities evolved in a similar way to end up with apparently comparable populations. At the time of the experiment, the results suggest that the microbial community inoculated for the bioaugmentation did not establish in the reactor but compensated for phosphorus accumulation until the acclimatisation of an endogenous microbial community arose.

Biodegradation, Environmental↗

PubMed2001

Biological denitrifying phosphorus removal in SBR: effect of added nitrate concentration and sludge retention time.

Optimizing anoxic biological phosphorus removal in the anaerobic-anoxic sequencing batch reactor (A2 SBR) was observed to depend on two parameters: the amount of added nitrate and the sludge retention time (SRT). The concentration of 120 mg N-NO3.l-1 in the anoxic medium and the SRT of 15 days were determined as optimal for a complete phosphorus removal in the A2 SBR. The reactor was supplied with synthetic wastewater containing 800 mg COD.l-1 acetic acid, 240 mg N-NH4.l-1 and 30 mg P-PO4.l-1. This study was completed by microscopic observations which revealed three morphological types of phosphate-accumulating bacteria (PAB).

Bacteria↗

PubMed2001

The anaerobic SBR process: basic principles for design and automation.

This study has determined the purification performance and the basic principles for the design of an anaerobic SBR (ASBR) to be used to treat wastewater generated in the food industries. Two ASBR's were set up and one fed with a slaughterhouse effluent at low concentration, the other with concentrated dairy wastewater. The maximum loading rate applied should not exceed 4.5 g of COD/L/day for the dilute effluent and 6 g of COD/L/day for the concentrated effluent. At higher loading rates, the reactors become difficult to operate, mainly because of sludge removal problems, and purification efficiency declines. A detailed study of the kinetics (TOC, VFA, rate of biogas production) throughout one treatment cycle led to the development of a simple control strategy based on the monitoring of the biogas production rate which was then applied to the reactor treating the dairy wastewater. After automation, the reactor worked free of problems at an average pollution load of 5.4 g of COD/L/day.

Abattoirs↗

PubMed2001

SBR as a relevant technology to combine anaerobic digestion and denitrification in a single reactor.

Two laboratory-scale sequencing batch reactors were used to study biological treatment of carbon and nitrogen in a new combined anaerobic-aerobic system. Piggery wastewater was used as a model effluent. The anaerobic reactor, fed with raw wastewater and nitrite/nitrate recycling from the aerobic reactor, carried out denitrification and anaerobic digestion of organic carbon. In the aerobic reactor, organic carbon removal and nitrification of ammonia to mainly nitrite occurred. Three recycle-to-influent ratios from 1 to 3 were tested. The higher the recycle-to-influent ratio, the lower the concentrations of nitrogen oxides in the final effluent. Carbon removal efficiency was quite stable, whatever the ratio. However, the effect of this ratio was attenuated because of denitrification in the aerobic reactor, which increased the performances of the process. The use of sequencing batch reactors was essential to apply the configuration proposed, in order to combine denitrification and methanogenesis that require antagonistic conditions.

Aerobiosis↗

PubMed2001

The SBR process: an efficient and economic solution for the treatment of wastewater at small cheesemaking dairies in the Jura mountains.

The small cheesemaking plants in the Jura Mountains (N.E. France) need self-contained solutions for treating their wastewater. Any solution must suit the characteristics of the effluent produced by these small production units and be simple, robust and financially feasible in regard to both capital and running costs. Given this context, a technique based on an SBR was recommended and the operations of the first treatment plant established on an industrial scale, monitored for the first year, with particular attention paid to operating constraints. The results obtained show that the SBR is, from a technical point of view, perfectly adapted to treating cheese production wastewater, with purification levels at 97.7% for total COD and 99.8% for BOD5. In the course of this study, it has been shown that the SBR process, thanks to its simple design and operation, can easily be run by the cheesemaker who will need to devote a minimum of time to it. Furthermore, with treatment costs of around 2 centimes (0.33 US cent) per litre of milk delivered, the SBR process meets perfectly both the technical and financial conditions for treatment as laid down by the Federation of Cooperatives of the Doubs and Jura départements.

Bioreactors↗

PubMed2000

Aerobic Denitrifiers Isolated from Diverse Natural and Managed Ecosystems.

Twenty-eight bacterial strains were isolated from an ecosystem adapted to fluctuating oxic-anoxic conditions. This ecosystem comprised a mixture of different natural and wastewater treatment environments. Among the 28 strains isolated, 10 exhibited aerobic denitrifying activity, i.e., co-respiration of oxygen and nitrate and simultaneous production of nitrite by 4 of them and of nitrogen gas by the remaining 6. Comparisons between the 16S rDNA sequences of the 10 strains showed that 3 of them were identical to M. aerodenitrificans, whereas RAPD profiles showed that the 3 strains were identical to each other but that they were different from M. aerodenitrificans. This implies that alternating aerobic-anoxic conditions allowed the isolation of a new strain of this aerobic denitrifier. Moreover, other denitrifying bacteria belonging to the genera Paracoccus, Thiobacillus, Enterobacter, Comamonas, and Sphingomonas were isolated in this way. These data imply that a wide variety of bacteria are able to carry out this type of metabolism. M. aerodenitrificans was also detected in methanogenic, denitrifying, nitrifying, phosphate removal, and activated sludge ecosystems by two-step PCR amplification. After 4 months of acclimation to oxic-anoxic phases, the strain was also detected in a canal and in a pond. This suggests that there is no specific natural ecological niche for aerobic denitrifiers but, as soon as selective pressure such as alternating aeration conditions is applied, this metabolism is amplified.

Journal Article↗

PubMed2000

Effect of dissolved oxygen and carbon-nitrogen loads on denitrification by an aerobic consortium.

Four samples of natural ecosystems and one sample from an activated sludge treatment plant were mixed together and progressively adapted to alternating aerobic/anoxic phases in the presence of nitrate in order to enrich the microflora in aerobic denitrifiers. Aerobic denitrifying performances of this mixed ecosystem at various dissolved oxygen concentrations and various carbon-nitrogen loads were evaluated and compared to those obtained with the aerobic denitrifier Microvirgula aerodenitrificans. The consortium and the pure strain exhibited an aerobic denitrifying activity at air saturation conditions (7 mg dissolved oxygen 1(-1)), i.e. there was co-respiration of the two electron acceptors with significant specific nitrate reduction rates. Dissolved oxygen concentrations had no influence on denitrifying performances above a defined threshold: 0.35 mg l(-1) for the consortium and 4.5 mg l(-1) for M. aerodenitrificans respectively. Under these thresholds, decreasing the dissolved oxygen concentrations enhanced the denitrifying activity of each culture. The higher the carbon and nitrogen loads, the higher the performance of the aerobic denitrifying ecosystem. However, for M. aerodenitrificans, the nitrate reduction percentage was affected more by variations in nitrogen load than in carbon load.

Aerobiosis↗

PubMed2000

Effect of culture conditions on the formation of struvite by Myxococcus xanthus.

The amount of struvite (MgNH4PO4 x 6H2O) produced by Myxococcus xanthus as well as the culture parameter values (pH, total phosphorus, total Kjeldahl nitrogen) were dependent on the culture medium used. Struvite formation started during the exponential phase and the maximum concentration was observed at the beginning of stationary growth phase. The addition of each medium component to the liquid culture influenced the amount of crystal produced. This amount did not depend on the pH increase during the culture period. The moment of the bacterial growth phase, at which each medium component was added, influenced the struvite formation.

Bacteriological Techniques↗

PubMed2000

Examination of two years of community dynamics in an anaerobic bioreactor using fluorescence polymerase chain reaction (PCR) single-strand conformation polymorphism analysis.

The structures of the bacterial and archaeal communities in an anaerobic digester were monitored over a 2 year period. The study was performed on a fluidized bed reactor fed with vinasse. The objective was to characterize the population dynamics over a long time period under constant environmental parameters. Total bacterial and archaeal populations were measured independently by fluorescence-based polymerase chain reaction (PCR) single-strand conformation polymorphism (SSCP) analysis using an automated DNA sequencer. With the current level of accuracy, the technique was able to monitor 45 bacterial and seven archaeal 16S rDNA molecules. The community dynamics were compared with molecular inventories of the microbial community based on 16S rDNA sequences done at the beginning of the study. The six archaeal and the 22 most frequent bacterial operational taxonomic units (OTUs) identified were associated with their SSCP peak counterparts. Overall, the data indicated that, throughout the period of the study, rapid significant shifts in the species composition of the bacterial community occurred, whereas the archaeal community remained relatively stable.

Archaea↗

PubMed2000

Ecological study of a bioaugmentation failure.

A nitrifying sequencing batch reactor was inoculated twice with the aerobic denitrifying bacterium Microvirgula aerodenitrificans and fed with acetate. No improvement was obtained on nitrogen removal. The second more massive inoculation was even followed by a nitrification breakdown, while at the same time, nitrification remained stable in a second reactor operated under the same conditions without bioaugmentation. Fluorescent in situ hybridization with rRNA-targeted probes revealed that the added bacteria almost disappeared from the reactor within 2 days, and that digestive vacuoles of protozoa gave strong hybridization signals with the M. aerodenitrificans-specific probe. An overgrowth of protozoa, coincident with the disappearance of free-living bacteria, was monitored by radioactive dot-blot hybridization only in the bioaugmented reactor. Population dynamics were analysed with a newly developed in situ quantification procedure of the probe-targeted bacteria. The nitrifying groups of bacteria decreased in a similar way in the bioaugmented and non-bioaugmented reactors. Other bacterial groups evolved differently. The involvement of different ecological parameters are discussed separately for each reactor. These results underline the importance of predator-prey interaction and illustrate the undesirable effects of massive bioaugmentation.

Animals↗

PubMed2000

Monitoring of activity dynamics of an anaerobic digester bacterial community using 16S rRNA polymerase chain reaction--single-strand conformation polymorphism analysis.

The influence of parameter changes on the bacterial community of a laboratory-scale anaerobic digester fed with glucose was investigated using a culture-independent approach based on single-strand conformation polymorphism (SSCP) analysis of total 16S rDNA and 16S rRNA amplification products. With the digester operating at steady state, the 16S rDNA SSCP patterns of the bacterial community showed eight peaks, whereas the 16S rRNA patterns showed six peaks with a very prominent one corresponding to a Spirochaetes-related bacterium. An acidic shock at pH 6 caused an increase in the 16S rRNA level of two Clostridium-related bacteria. After a 1 week starvation period, the major bacteria present reverted to a basal 16S rRNA level proportional to their 16S rDNA level. Starvation revealed the presence of a previously undetected peak whose corresponding sequence was deeply branched into the low G+C Gram-positive bacteria phylum. Twenty-four hours after a spiked addition to the starved digester community of starch, glucose, lactate or sulphate, an upsurge in several new 16S rRNA-derived peaks was observed. Thus, the perturbation approach combined with 16S rRNA analysis revealed bacteria that had not been detected through 16S rDNA analysis.

Bacteria, Anaerobic↗

PubMed2000

Investigations on the changes in anaerobic biodegradability and biotoxicity of an industrial microbial biomass induced by a thermochemical pretreatment.

In the anaerobic digestion of solid wastes, hydrolysis is the rate limiting step and physicochemical pretreatment is often required to promote solubilization of organic matter. As an illustration, anaerobic digestion was limited by the substrate solubilization step during continuous cultures performed with an industrial microbial biomass. In optimal operating conditions determined for the hydrolysis-acidogenesis step (T = 35 degrees C; pH = 8.5; OLR = 5.4 g COD/l.d) 50.6% COD solubilization was achieved. A thermochemical pretreatment based on sodium hydroxide addition, was used in order to enhance COD solubilization. Optimal conditions for COD solubilization were pH = 12, T = 140 degrees C for 30 minutes. In these conditions, 70% COD solubilization was achieved. However, anaerobic biodegradability of the pretreated substrate was not improved and remained near 40%. The poor anaerobic biodegradability performances were attributed to the soluble molecules generated during the thermochemical pretreatment that were refractory and/or inhibitory to anaerobic microorganisms. Fractionation of the soluble pretreated microbial biomass by two methods (treatment with adsorbent resins and precipitation by pH adjustment) demonstrated that high molecular weight compounds (> 100 kDa) are involved in the poor biodegradability and in the biotoxicity observed. Partial decolorization through resin use and acid precipitation remove these compounds. The consequence of their removal was an increase of the production of biogas.

Acetates↗

PubMed1999

Polyphosphate-accumulating and denitrifying bacteria isolated from anaerobic-anoxic and anaerobic-aerobic sequencing batch reactors

In this study, phosphate-accumulating bacteria achieved complete phosphate removal in two different systems: an anaerobic-anoxic sequencing batch reactor and an anaerobic-aerobic sequencing batch reactor. This result shows that phosphate-accumulating bacteria in the A2 SBR can use nitrate as terminal electron acceptor instead of oxygen. Phosphate-accumulating bacteria accumulated phosphate with a rates between 30 and 70 mg P/L/h in the A/O SBR and between 15 and 32 mg P/L/h in the A2 SBR. Twenty denitrifying isolates were screened from A2 SBR and nine from A/O SBR. Identification of these isolates by the Biolog system and the API 20 NE identification kit revealed that the most active denitrifiers in both SBRs reactors were species of Ochrobactrum, Pseudomonas, Corynebacterium, Agrobacterium, Aquaspirillum, Haemophilus, Xanthomonas, Aeromonas, and Shewanella. The most active phosphate accumulating and denitrifying bacteria were identified as Agrobacterium tumefaciens B, Aquaspirillum dispar, and Agrobacterium radiobacter. This study showed that the active phosphate accumulating-bacteria were also the most efficient denitrifying bacteria in both reactors.

Journal Article↗

PubMed1999

Kinetic behavior of some polyphosphate-accumulating bacteria isolates in the presence of nitrate and oxygen.

This paper studies the phosphate uptake by pure cultures of Aeromonas hydrophila, Klebsiella oxytoca, Agrobacterium tumefaciens, and Aquaspirillum dispar in the presence of both nitrate and oxygen. It is shown that species were able to respire both electron acceptors for phosphate accumulation. A. tumefaciens and A. dispar accumulated overall phosphate both in oxic and anoxic culture conditions, whereas A. hydrophila and K. oxytoca eliminated overall phosphate only in oxic conditions. A. dispar was able to remove phosphate by reducing oxygen and nitrate simultaneously with the production of dinitrogen gas. The anoxic denitrification observed in the cultures of adapted and nonadapted cells to nitrate showed that only A. dispar have a denitrification rate superior when the cells were adapted to nitrate.

Aeromonas↗

PubMed1998

Mixing and phase hold-ups variations due to gas production in anaerobic fluidized-bed digesters: influence on reactor performance

The influence of mixing and phase hold-ups on gas-producing fluidized-bed reactors was investigated and compared with an ideal flow reactor performance (CSTR). The liquid flow in the anaerobic fluidized bed reactor could be described by the classical axially dispersed plug flow model according to measurements of residence time distribution. Gas effervescence in the fluidized bed was responsible for bed contraction and for important gas hold-up, which reduced the contact time between the liquid and the bioparticles. These results were used to support the modeling of large-scale fluidized-bed reactors. The biological kinetics were determined on a 180-L reactor treating wine distillery wastewater where the overall total organic carbon uptake velocity could be described by a Monod model. The outlet concentration and the concentration profile in the reactor appeared to be greatly influenced by hydrodynamic limitations. The biogas effervescence modifies the mixing characteristics and the phase hold-ups. Bed contraction and gas hold-up data are reported and correlated with liquid and gas velocities. It is shown that the reactor performance can be affected by 10% to 15%, depending on the mode of operation and recycle ratio used. At high organic loading rates, reactor performance is particularly sensitive to gas effervescence effects. Copyright 1998 John Wiley & Sons, Inc.

Journal Article↗