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PubMed2000

An evaluation of sampling and laboratory procedures for determination of heterotrophic plate counts in dental unit waterlines.

BACKGROUND: The high numbers of heterotrophic microorganisms that have been cultured from dental unit waterlines (DUWs) have raised concern that this water may exceed suggested limits for heterotrophic plate counts (HPCs). The main purpose of this investigation was to examine HPC variability in DUWs and to examine in detail the effect of laboratory processing of water samples on HPC values. METHODS: Water samples were collected from dental offices either at the beginning of or during the clinic day and were transported to the laboratory, where they were analyzed. RESULTS: Measuring HPC levels within an office would involve testing all units, because significant differences were found between units connected to the same municipal water supply. Within a unit, the average microbial count from high-speed lines was approximately twice the average count from air/water lines. The laboratory processing of water samples significantly affected the numbers of heterotrophic microorganisms that were recovered. Incubation temperature, time and media, as well as neutralization of residual chlorine, all had significant effects on the HPC values. However, no significant differences in microbial counts were found between samples plated with the spread plate method on R2A agar and those plated with the pour plate method with Plate Count Agar. CONCLUSIONS: Dental organizations have suggested target limits in terms of numbers of heterotrophic microorganisms recovered in water from dental units, but standards for laboratory handling must be established as well. A protocol for sample collection and laboratory handling is proposed.

Bacteriological Techniques↗
PubMedDate not supplied

[Theoretical evaluation of necessity of carbon dioxide assimilation by microorganisms during growth on various substrates].

The biological role of exogenous carbon dioxide during substrate assimilation with a various degree of reductivity is evaluated. The investigation of metabolic pathways of carbon dioxide incorporation into the metabolic processes of methaneoxidizing bacteria shows that the HCO3- ion assimilation is catalyzed by phosphoenolpyruvate carboxylase and in certain strains also by the key enzyme of autotrophic pathway of the carbon dioxide assimilation, ribulose-1,5-diphosphate carboxylase. The theoretical calculations and experimental studies indicate that exogenous carbon dioxide is a necessary participant of the metabolic processes of methane or methanol assimilation. It is also an acceptor of the excess electrons of these compounds. It is the degree of reductivity of the substrate metabolized that determines the activity of the exogenous carbon dioxide fixation by microorganisms. The carbon dioxide fixation by heterotrophic microorganisms must be considered, therefore, as a process which is mostly due to the elementary composition of the source of carbon under conversion.

Bacteria↗
PubMed2005

Characteristics and adaptability of iron- and sulfur-oxidizing microorganisms used for the recovery of metals from minerals and their concentrates.

Microorganisms are used in large-scale heap or tank aeration processes for the commercial extraction of a variety of metals from their ores or concentrates. These include copper, cobalt, gold and, in the past, uranium. The metal solubilization processes are considered to be largely chemical with the microorganisms providing the chemicals and the space (exopolysaccharide layer) where the mineral dissolution reactions occur. Temperatures at which these processes are carried out can vary from ambient to 80 degrees C and the types of organisms present depends to a large extent on the process temperature used. Irrespective of the operation temperature, biomining microbes have several characteristics in common. One shared characteristic is their ability to produce the ferric iron and sulfuric acid required to degrade the mineral and facilitate metal recovery. Other characteristics are their ability to grow autotrophically, their acid-tolerance and their inherent metal resistance or ability to acquire metal resistance. Although the microorganisms that drive the process have the above properties in common, biomining microbes usually occur in consortia in which cross-feeding may occur such that a combination of microbes including some with heterotrophic tendencies may contribute to the efficiency of the process. The remarkable adaptability of these organisms is assisted by several of the processes being continuous-flow systems that enable the continual selection of microorganisms that are more efficient at mineral degradation. Adaptability is also assisted by the processes being open and non-sterile thereby permitting new organisms to enter. This openness allows for the possibility of new genes that improve cell fitness to be selected from the horizontal gene pool. Characteristics that biomining microorganisms have in common and examples of their remarkable adaptability are described.

Journal Article↗
PubMed1997

Health impacts of large releases of radionuclides. Roles of micro-organisms in the environmental fate of radionuclides.

Micro-organisms play important roles in the environmental fate of radionuclides in both aquatic and terrestrial ecosystems, with a multiplicity of physico-chemical and biological mechanisms effecting changes in mobility and speciation. Physico-chemical mechanisms of removal, which may be encompassed by the general term 'biosorption', include adsorption, ion exchange and entrapment. These are features of living and dead organisms as well as their derived products. In living cells biosorptive processes can be directly and indirectly influenced by metabolism, and may be reversible and affected by changing environmental conditions. Metabolism-dependent mechanisms of radionuclide immobilization include metal precipitation as sulfides, sequestration by metal-binding proteins and peptides, and transport and intracellular compartmentation. Chemical transformations of radionuclide species, particularly by reduction, can result in immobilization. Microbial processes involved in solubilization include autotrophic and heterotrophic leaching, complexation by siderophores and other metabolites, and chemical transformations. Such mechanisms are important components of natural biogeochemical cycles for radionuclides and should be considered in any analyses of environmental radionuclide contamination. Several micro-organism-based biotechnologies, e.g. those based on biosorption or precipitation, are of potential use for the treatment of radionuclide contamination.

Adsorption↗
PubMed2004

Biotechnological production and applications of the omega-3 polyunsaturated fatty acid docosahexaenoic acid.

Docosahexaenoic acid (DHA) is a polyunsaturated fatty acid composed of 22 carbon atoms and six double bonds. Because the first double bond, as counted from the methyl terminus, is at position three, DHA belongs to the so-called omega-3 group. In recent years, DHA has attracted much attention because of its beneficial effect on human health. At present, fish oil is the major source of DHA, but alternatively it may be produced by use of microorganisms. Marine microorganisms may contain large quantities of DHA and are considered a potential source of this important fatty acid. Some of these organisms can be grown heterotrophically on organic substrates without light. These processes can be well controlled and DHA with constant quality can be produced all year round. This paper reviews recent advances in the biotechnological production of DHA by marine microorganisms.

Animals↗
PubMed2004

Diversity and classification of mycorrhizal associations.

Most mycorrhizas are 'balanced' mutualistic associations in which the fungus and plant exchange commodities required for their growth and survival. Myco-heterotrophic plants have 'exploitative' mycorrhizas where transfer processes apparently benefit only plants. Exploitative associations are symbiotic (in the broad sense), but are not mutualistic. A new definition of mycorrhizas that encompasses all types of these associations while excluding other plant-fungus interactions is provided. This definition recognises the importance of nutrient transfer at an interface resulting from synchronised plant-fungus development. The diversity of interactions between mycorrhizal fungi and plants is considered. Mycorrhizal fungi also function as endophytes, necrotrophs and antagonists of host or non-host plants, with roles that vary during the lifespan of their associations. It is recommended that mycorrhizal associations are defined and classified primarily by anatomical criteria regulated by the host plant. A revised classification scheme for types and categories of mycorrhizal associations defined by these criteria is proposed. The main categories of vesicular-arbuscular mycorrhizal associations (VAM) are 'linear' or 'coiling', and of ectomycorrhizal associations (ECM) are 'epidermal' or 'cortical'. Subcategories of coiling VAM and epidermal ECM occur in certain host plants. Fungus-controlled features result in 'morphotypes' within categories of VAM and ECM. Arbutoid and monotropoid associations should be considered subcategories of epidermal ECM and ectendomycorrhizas should be relegated to an ECM morphotype. Both arbuscules and vesicles define mycorrhizas formed by glomeromycotan fungi. A new classification scheme for categories, subcategories and morphotypes of mycorrhizal associations is provided.

Fungi↗
PubMed2006

Reduction of the pea ferredoxin-NADP(H) reductase catalytic efficiency by the structuring of a carboxyl-terminal artificial metal binding site.

Ferredoxin (flavodoxin)-NADP(H) reductases (FNRs) are ubiquitous flavoenzymes that deliver NADPH or low-potential one-electron donors (ferredoxin, flavodoxin, and adrenodoxin) to redox-based metabolisms in plastids, mitochondria, and bacteria. The FNRs from plants and most eubacteria constitute a unique family, the plant-type ferredoxin-NADP(H) reductases. Plastidic FNRs are quite efficient at sustaining the demands of the photosynthetic process. At variance, FNRs from organisms with heterotrophic metabolisms or anoxygenic photosynthesis display turnover numbers that are 20-100-fold lower than those of their plastidic and cyanobacterial counterparts. To gain insight into the FNR structural features that modulate enzyme catalytic efficiency, we constructed a recombinant FNR in which the carboxyl-terminal amino acid (Tyr308) is followed by an artificial metal binding site of nine amino acids, including four histidine residues. This added structure binds Zn2+ or Co2+ and, as a consequence, significantly reduces the catalytic efficiency of the enzyme by decreasing its kcat. The Km for NADPH and the Kd for NADP+ were increased 2 and 3 times, respectively, by the addition of the amino acid extension in the absence of Zn2+. Nevertheless, the structuring of the metal binding site did not change the Km for NADPH or the Kd for NADP+ of the FNR-tail enzyme. Our results provide experimental evidence which indicates that mobility of the carboxyl-terminal backbone region of the FNR, mainly Tyr308, is essential for obtaining an FNR enzyme with high catalytic efficiency.

Amino Acid Sequence↗
PubMed2003

Enrichment from activated sludges of aerobic mixed cultures capable to degrade vinyl chloride (VC) as the sole carbon source.

Two microbial cultures able to degrade high concentrations of VC as the sole carbon source have been obtained by enrichment from activated sludge. The cultures began consuming VC (0.02 mmol l(-1)) only after a long initial acclimation period (1-2 months). After then the concentration of VC was gradually increased (from 0.02 to 0.8 mmol l(-1)) and the cultures were able to maintain VC degrading ability for long time (over 500 days). VC-degrading biomass in the two cultures was characterized by low specific maximum growth rates (0.19-0.21 d(-1)) compared to heterotrophic organisms typically present in activated sludge processes. Monod half-saturation constant was rather low (0.7-1.6 mg VC l(-1)) indicating that it is possible to effectively remove VC to low residual concentrations. The cultures were highly sensitive to even short periods of VC lack (with quick decrease of VC degradation rates) whereas they were not to sudden load increases (up to 3.4 mmol l(-1)). After being cultured with only ethene as the sole carbon and energy source, the cultures kept the ability of degrading VC. Possibility of maintaining the mixed cultures on non-toxic ethene, without loosing VC degradation ability, is very promising for bioaugmentation treatments.

Bacteria, Aerobic↗
PubMed1995

Euglena gracilis chloroplast psbB, psbT, psbH and psbN gene cluster: regulation of psbB-psbT pre-mRNA processing.

A 2.4 kb region of the Euglena gracilis chloroplast genome containing the genes psbT, psbH and psbN was characterized. The mRNAs transcribed from psbB, psbT, psbH and psbN were analyzed by northern hybridization, S1 nuclease protection analysis and primer extension RNA sequencing. The gene pairs psbB-psbT and psbH-psbN are cotranscribed from opposite strands. The 5' end of the psbN-psbH transcript and the intercistronic cleavage sites between psbB-psbT and psbN-psbH were determined. The extent of psbB-psbT intercistronic cleavage is greater during photoautotrophic than heterotrophic growth and thus may be developmentally regulated. Processing is absent in the non-photosynthetic E. gracilis mutant Y9Z1NaL.

Animals↗
PubMed2004

Effect of Alcaligenes faecalis on nitrous oxide emission and nitrogen removal in three phase fluidized bed process.

Nitrous oxide (N2O), one of the greenhouse effect gases, has not been known that how much N2O is produced from municipal wastewater treatment and what its management should be. In this study, for controlling nitrous oxide emission and removing nitrogen from municipal wastewater, we experimented the three phase fluidized bed process equipped with draft tube along with immobilized Alcaligenes faecalis, a typical heterotrophic nitrifer and a predominant genus. Also we evaluated the optimum treatment condition of the three phase fluidized bed process for emitting nitrous oxide. The results of this study showed that the three phase fluidized bed process was more effective than the activated sludge process for controlling nitrous oxide emission and removing nitrogen. Increasing amount of A. faecalis in reactor should be encouraged for controlling nitrous oxide emission and removing nitrogen. In addition, the activated sludge process using immobilized A. faecalis as a carrier had more nitrogen removal efficiency than conventional activated sludge process. The accumulation of NO2-N, NO3-N resulted in high N2O emission. Therefore, we suggested that it is necessary to reduce NO2-N and NO3-N for both reducing N2O emission and improving nitrogen removal.

Alcaligenes↗
PubMed1983

Comparison between direct methods for determination of microbial cell volume: electron microscopy and electronic particle sizing.

Size frequency distributions of different phototrophic and heterotrophic microorganisms were determined by means of scanning and transmission electron microscopy and electronic particle sizing. Statistically significant differences existed among the three techniques used in this study. Cells processed for electron microscopy showed lower mean cellular volumes than those processed for electronic particle sizing, reflecting a shrinkage by factors ranging from 1.1 to 6.2 (mean, 2.3). Processing of cells for scanning electron microscopy caused higher shrinkage than processing for transmission electron microscopy. Shrinkage was dependent neither on the size nor on the cell wall type of the microorganism. When processed for scanning electron microscopy, phototrophic bacteria were strongly shrunken, whereas heterotrophic microorganisms were less affected. A direct relationship existed among phototrophic bacteria between percentage of shrinkage and specific pigment content. This was probably a consequence of the pigment extraction by organic solvents during the dehydration process, previous to the critical point drying, necessary to examine the specimens under the scanning electron microscope.

Bacteria↗
PubMed2005

Heterotrophic fixation of CO(2) in soil.

The occurrence of heterotrophic CO(2) fixation by soil microorganisms was tested in several mineral soils differing in pH and two artificial soils (a mixture of silica sand, alfalfa powder, and nutrient medium inoculated with a soil suspension). Soils were incubated at ambient ( approximately 0.05 vol%) and elevated ( approximately 5 vol%) CO(2) concentrations under aerobic conditions for up to 21 days. CO(2) fixation was detected using either a technique for determining the natural abundance of (13)C or by measuring the distribution of labeled (14)C-CO(2) in soil and bacteria. The effects of elevated CO(2) on microbial biomass (direct counts, chloroform fumigation extraction method), composition of microbial community (phospholipid fatty acids), microbial activity (respiration, dehydrogenase activity), and turnover rate were also measured. Heterotrophic CO(2) fixation was proven in all soils under study, being higher in neutral soils. The main portion of the fixed CO(2) (98-99%) was found in extracellular metabolites while only approximately 1% CO(2) was incorporated into microbial cells. High CO(2) concentration always induced an increase in microbial activity, changes in the composition of the microbial community, and a decrease in microbial turnover. The results suggest that heterotrophic CO(2) fixation could be a widespread process in soils.

Bacteria↗
PubMed2005

Biodiesel production from heterotrophic microalgal oil.

The present study introduced an integrated method for the production of biodiesel from microalgal oil. Heterotrophic growth of Chlorella protothecoides resulted in the accumulation of high lipid content (55%) in cells. Large amount of microalgal oil was efficiently extracted from these heterotrophic cells by using n-hexane. Biodiesel comparable to conventional diesel was obtained from heterotrophic microalgal oil by acidic transesterification. The best process combination was 100% catalyst quantity (based on oil weight) with 56:1 molar ratio of methanol to oil at temperature of 30 degrees C, which reduced product specific gravity from an initial value of 0.912 to a final value of 0.8637 in about 4h of reaction time. The results suggested that the new process, which combined bioengineering and transesterification, was a feasible and effective method for the production of high quality biodiesel from microalgal oil.

Biotechnology↗
PubMed2002

Calorimetry: a tool for assessing microbial activity under aerobic and anoxic conditions.

For many years, calorimetric measurements have been used for understanding, modelling, controlling, and optimising chemical reactions. Calorimetry could be as well utilised to investigate biological processes, which however, involve very small amount of heat and therefore require very sensitive instruments. For this purpose, a Mettler Toledo RCI (Reaction calorimeter) was modified, changing both hardware and software, increasing its resolution up to 5 10m W/l. Such sensitivity allows the monitoring of aerobic and anoxic processes. This paper points out the excellent agreement between calorimetric and respirometric data, obtained simultaneously under aerobic conditions using activated sludge from a lab-scale scale reactor. Heat production rate can be directly converted in oxygen uptake rate by means of a correlation factor, whose value is approximately the same for all aerobic respiratory metabolisms. Taking into account this factor, calorimetric data were introduced in a chemical oxygen demand based model and processed for the estimation of kinetic parameters of heterotrophic biomass. Aerobic heterotrophic, denitrifying, and autotrophic nitrifying activity were determined by specific calorimetric tests. The effect of potentially toxic or inhibitory substances on the activity of all microbial communities was as well pointed out in these measurements.

Bacteria, Aerobic↗
PubMed2002

Biotransformation of 2,4,6-trinitrotoluene in a continuous-flow Anabaena sp. system.

Reductive transformation of 2,4,6-trinitrotoluene (TNT) was observed in a continuous-flow system of Anabaena sp. operated for 33 d with a 5.7 d hydraulic retention time and a range of influent TNT concentrations of 1-58 mg/l. The TNT removal efficiency of the continuous-flow system at the highest influent TNT concentration of 58 mg/l was 96.7 +/- 1.7% (mean +/- 95% confidence interval). Culture chlorosis and growth inhibition were not observed during this study. The pseudo-first order TNT transformation rate constant values corresponding to the system performance range (0.14-0.46/h) were lower than the values previously recorded for batch Anabaena sp. cultures with less than 10 mg/l initial TNT concentrations, possibly due to an inhibition of the TNT transformation process by either TNT and/or TNT transformation products. Heterotrophic bacterial populations developed in the continuous-flow Anabaena sp. cultures also transformed TNT, but at a much lower rate than the Anabaena sp. Less than 1% of the overall TNT transformation observed in the continuous-flow system was attributed to the heterotrophic bacterial populations. The only TNT reduction products identified in both the culture media and in biomass extracts were azoxytetranitrotoluene isomers and low levels of aminodinitrotoluene isomers. TNT and TNT transformation products identified in the culture effluent and the biomass extract accounted for only about 24% of the TNT added to the system (on a molar basis). Production of soluble, polar metabolites, uptake, partial mineralization and/or sequestration of TNT and its transformation products by Anabaena may be responsible for the relatively low contaminant recovery and mass balance observed in this study.

Anabaena↗
PubMed1996

Influence of microorganisms on the environmental fate of radionuclides.

Microorganisms have a significant influence on the environmental fate of radionuclides in aquatic and terrestrial ecosystems with a multiplicity of physico-chemical and biological mechanisms effecting changes in mobility and speciation. Physico-chemical mechanisms of removal include association with extracellular materials, metabolites and cell walls which are features of living and dead organisms. In living cells, some physico-chemical processes are reversible, influenced by metabolism and changing environmental conditions. Metabolism-dependent mechanisms of radionuclide immobilization include sulphide precipitation, transport and intracellular compartmentation and/or sequestration by proteins and peptides. In addition, chemical reduction to less soluble forms can result in immobilization. Microbial processes involved in radionuclide solubilization include autotrophic and heterotrophic leaching, and complexation by siderophores and other metabolites. Such mechanisms are important components of biogeochemical cycles for radionuclides and should be considered in any analyses of environmental radionuclide contamination. In addition, several microorganism-based biotechnologies are receiving interest as potential treatment methods.

Absorption↗
PubMed2006

In situ analysis of nitrogen fixation and metabolic switching in unicellular thermophilic cyanobacteria inhabiting hot spring microbial mats.

Genome sequences of two Synechococcus ecotypes inhabiting the Octopus Spring microbial mat in Yellowstone National Park revealed the presence of all genes required for nitrogenase biosynthesis. We demonstrate that nif genes of the Synechococcus ecotypes are expressed in situ in a region of the mat that varies in temperature from 53.5 degrees C to 63.4 degrees C (average 60 degrees C); transcripts are only detected at the end of the day when the mat becomes anoxic. Nitrogenase activity in mat samples was also detected in the evening. Hitherto, N2 fixation in hot spring mats was attributed either to filamentous cyanobacteria (not present at >50 degrees C in these mats) or to heterotrophic bacteria. To explore how energy-generating processes of the Synechococcus ecotypes track natural light and O2 conditions, we evaluated accumulation of transcripts encoding proteins involved in photosynthesis, respiration, and fermentation. Transcripts from photosynthesis (cpcF, cpcE, psaB, and psbB) and respiration (coxA and cydA) genes declined in the evening. In contrast, transcripts encoding enzymes that may participate in fermentation fell into two categories; some (ldh, pdhB, ald, and ackA) decreased in the evening, whereas others (pflB, pflA, adhE, and acs) increased at the end of the day and remained high into the night. Energy required for N2 fixation during the night may be derived from fermentation pathways that become prominent as the mat becomes anoxic. In a broader context, our data suggest that there are critical regulatory switches in situ that are linked to the diel cycle and that these switches alter many metabolic processes within the microbial mat.

Energy Metabolism↗
PubMed2002

Who eats what? Classifying microbial populations based on diurnal profiles of rRNA levels.

Identifying the relationships between various bacterial populations and the substrates they consume is central to the understanding of population dynamics and to the development of process control in activated sludge. However, linking a heterotrophic population to its activity in situ is difficult because ribosomal RNA (rRNA) techniques, while allowing the rapid identification of populations, provide little information about their heterotrophic activity. Activated sludge models describe biodegradation kinetics by classifying substrates into two types: readily and slowly degradable substrates. Assuming that bacterial populations specialize in degrading one type of substrate, their growth rate should be affected differently if the COD loading rate varies diurnally as for a municipal activated sludge system. Modeling results suggested that the growth rates of populations consuming readily degradable substrates vary according to variations in COD loading rate. On the other hand, the growth rates of populations consuming slowly degradable substrates do not change despite the variation in COD loading rate. Since the cellular rRNA level is positively correlated with the growth rate, we hypothesized that the rRNA levels of some populations in municipal activated sludge should increase throughout the day, while they should stay constant for other populations. This hypothesis was verified by monitoring the rRNA level of Acinetobacter (a model population consuming readily degradable substrates) and Gordonia (a model population consuming slowly degradable substrates) in the mixed liquor of a full-scale municipal activated sludge reactor for three weeks.

Bacteria↗