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Epidemiologic typing of nosocomial microorganisms.

Hospital epidemiologists often rely upon "typing" of microorganisms to help determine their genetic relatedness. Most general clinical microbiology laboratories can determine biologic profiles (biotypes) and antimicrobial susceptibility patterns (antibiograms) of bacteria commonly isolated from specimens; occasionally serologic typing (serotype) is also performed. Special interest laboratories can provide serologic typing, bacteriophage susceptibility patterns (phage typing), bacteriocin production patterns, bacteriocin susceptibility patterns, plasmid analyses and chromosomal DNA analyses for a variety of bacteria, mycobacteria and fungi of nosocomial interest. Such laboratories can also provide serologic typing, restriction enzyme analyses and other special studies of viruses and related microorganisms. A useful and effective "typing" system should be (1) standardized, (2) reproducible, (3) sensitive, (4) stable, (5) available, (6) inexpensive, (7) applicable to a wide range of microorganisms, and (8) field tested in conjunction with epidemiologic investigation. Results should be reported in a standard manner with some discussion of the implications and limitations of the reported results. We suggest that a registry of typing methods be established to facilitate application of available methods to appropriate epidemiologic investigations.

Bacteria↗

Production of prostaglandin E2 by human amnion in vitro in response to addition of media conditioned by microorganisms associated with chorioamnionitis and preterm labor.

To examine the potential role of bacterial infection in the cause of spontaneous preterm labor, human amnion cells in tissue culture were exposed to medium conditioned by culturing each of 21 microorganisms previously found in association with chorioamnionitis and preterm labor. At a final concentration of 0.1% bacterial conditioned medium, a significant stimulation of prostaglandin E2 production from amnion cells was observed for this range of microorganisms. Conditioned medium obtained from culturing Bacteroides fragilis caused a dose-related increase in prostaglandin production, final concentrations of 0.02% to 0.1% being stimulatory but greater concentrations (0.1% to 10%) causing a progressive inhibition of prostaglandin synthesis. A similar concentration-related response in which stimulation was followed by inhibition occurred on addition of increasing concentrations of phospholipase A2 to amnion cells. These data suggest that bacterial phospholipase may release arachidonic acid from amnion leading to prostaglandin E2 synthesis, but excessive addition of phospholipase and consequent increased arachidonic acid availability may give rise to substrate inhibition of cyclooxygenase enzyme and inhibit prostaglandin E2 synthesis. Overall it appears that a wide variety of microorganisms associated with preterm labor may secrete phospholipase, which liberates amnion arachidonic acid for conversion to the oxytocic agent prostaglandin E2.

Amnion↗

Acid-base pH curves in vitro with mixtures of pure cultures of human oral microorganisms.

Pure cultures of microorganisms commonly found in supragingival plaque were incubated alone and in combinations to determine the bacterial contribution to the pH-fall-pH-rise that is the central characteristic of the Stephan-curve pH change seen in plaque in vivo after brief exposure to a sugar solution. To avoid the complicating conditions of saliva flow and plaque diffusion, experiments were done with bacterial suspensions in incubations in vitro. In an initial experimental series where each microorganism was incubated only with glucose, all but a few produced the initial pH fall. Some also showed a subsequent small, sharp rise in the pH which then quickly levelled off; this was due to metabolism of endogenous substrate accumulated by most microorganisms during their growth in culture. When arginolytic and non-arginolytic bacteria were each then incubated with both glucose and arginine present (the glucose substrate to stimulate a pH fall and the arginine to stimulate a pH rise), the non-arginolytic gave a progressively more acidic pH response with progressive increase in the cell concentration, whereas the arginolytic bacteria produced a much smaller and variable pH decrease with similar cell concentration increase. Mixing pure cultures of either arginolytic or non-arginolytic bacteria gave acid-base pH responses similar to those of their respective pure cultures, whereas mixing arginolytic with non-arginolytic bacteria resulted in an approximate averaging of their different curves. The organisms present in highest proportion in a mixture had the greatest effects. The outcome of mixing the most numerous streptococcal and actinomyces species found normally in supragingival plaque indicated that the well-established difference in the acidity level of the Stephan pH response of caries-active and caries-inactive plaques could be due to differences in the proportions of their arginolytic and non-arginolytic members.

Arginine↗

Microorganism track reconstruction: an image processing approach.

This paper presents an automatic system for the analysis of microorganism behaviour. The movements of free swimming microorganisms are videotaped by means of a television camera mounted on a microscope. The analysis is performed off-line by digitizing the video signal through the use of an image processor unit. Microorganism tracks are reconstructed spatially and chronologically by means of image processing techniques. From these tracks cell movement parameters are obtained. The results of our experiment in testing photoinduced movements follow.

Algorithms↗

Tarnish of dental alloys by oral microorganisms.

Five dental alloys, on exposure to blood and chocolate media with and without inoculated microorganisms, showed varying degrees of tarnish. The results indicated a composition-dependent tarnish behavior of alloys in microorganism-inoculated media, indicating a potential role for the oral microorganisms in inducing clinically observed tarnish of dental alloys. Actinomyces viscosus and periodontal pocket specimens show a similarity in their activity to induce tarnish in base metal-containing dental alloys.

Actinomyces viscosus↗

Physiology of food poisoning microorganisms and the major problems in food poisoning control.

There remains considerable public concern regarding the current high level of food poisoning disease in Europe and the fact that, year by year, it continues to rise rather than fall. At the same time, there are strong and increasing demands from consumers for foods that are more convenient, fresher, more natural, less heavily processed (e.g. 'REPFEDS' and 'Sous Vide' foods, mildly heated and distributed at chill temperatures; Lund and Notermans, 1992), less heavily preserved (e.g. less acid, less salt, less sugar; Gould, 1995) and less reliant on additive preservatives than hitherto (e.g. sulphite, nitrite, organic acids and esters; Russell and Gould, 1991). Most of these trends result in a general reduction in the intrinsic preservation of foods. Furthermore, many food poisoning microorganisms escape the attention of preservation techniques altogether, reaching the consumer more or less directly from contaminated foods, most often foods of animal origin. It has therefore been argued that a substantial reduction in food poisoning in the near future will be difficult to achieve unless we obtain a greatly improved understanding of the physiology of the most important target organisms (Knochel and Gould, 1995). This knowledge must then be exploited in ways which effectively improve our means for the control of these hazards and reduce the risk to the consumer. A three year AAIR Concerted Action Programme (PL920630: 'Physiology of Food Poisoning Microorganisms') was therefore initiated in 1992 in order to bring together research groups working on the physiology and related aspects of food poisoning microorganisms. The principal objectives of the programme were: 1. To determine the physiological, biochemical and genetical bases of the organisms' survival of and responses to food-relevant stresses; 2. to determine the physiological and genetical factors influencing infectivity and toxinogenesis; 3. to understand the physiological bases of those synergistic systems that are already empirically applied or that have future potential; 4. to make a wide range of modern techniques in which particular members have expertise more widely available. As can be read in the subsequent contributions to this special issue, the area is a fruitful one for microbiological research and the Programme has been successful in bringing together disparate strands of the topic. It has also highlighted areas where this scientific knowledge may be better exploited in improving the microbiological safety of foods for the consumer.

Bacterial Physiological Phenomena↗

Microorganisms in honey.

Knowledge of the moisture and temperature conditions influencing growth of microorganisms in honey has long been used to control the spoilage of honey. However, the need for additional microbiological data on honey will increase as new technologies for, and uses of honey develop. Microorganisms in honey may influence quality or safety. Due to the natural properties of honey and control measures in the honey industry, honey is a product with minimal types and levels of microbes. Microbes of concern in post-harvest handling are those that are commonly found in honey (i.e., yeasts and spore-forming bacteria), those that indicate the sanitary or commercial quality of honey (i.e., coliforms and yeasts), and those that under certain conditions could cause human illness. Primary sources of microbial contamination are likely to include pollen, the digestive tracts of honey bees, dust, air, earth and nectar, sources which are very difficult to control. The same secondary (after-harvest) sources that influence any food product are also sources of contamination for honey. These include air, food handlers, cross-contamination, equipment and buildings. Secondary sources of contamination are controlled by good manufacturing practices. The microbes of concern in honey are primarily yeasts and spore-forming bacteria. Total plate counts from honey samples can vary from zero to tens of thousands per gram for no apparent reason. Most samples of honey contain detectable levels of yeasts. Although yeast counts in many honey samples are below 100 colony forming units per gram (cfu/g), yeasts can grow in honey to very high numbers. Standard industry practices control yeast growth. Bacterial spores, particularly those in the Bacillus genus, are regularly found in honey. The spores of C. botulinum are found in a fraction of the honey samples tested-normally at low levels. No vegetative forms of disease-causing bacterial species have been found in honey. Bacteria do not replicate in honey and as such high numbers of vegetative bacteria could indicate recent contamination from a secondary source. Certain vegetative microbes can survive in honey, at cool temperatures, for several years. However, honey has anti-microbial properties that discourage the growth or persistence of many microorganisms. Typically, honey can be expected to contain low numbers and a limited variety of microbes. A routine microbiological examination of honey might include several different assays. A standard plate count provides general information. Specialized tests, such as a count of yeasts and an assay for bacterial spore-formers, may also be useful. An indicator of sanitary quality as provided by coliform counts might be included. Additional tests, to explain unusually high counts or address a certain problem, may be needed. The use of honey in products that receive no or limited heat treatment may require additional tests. More information on the source and control of microbes in honey is needed to answer the concerns currently facing the industry.

Anti-Bacterial Agents↗

Distribution of adenosine 5'-triphosphate (ATP)-dependent hexose kinases in microorganisms.

A systematic study of adenosine triphosphate (ATP)-dependent hexose kinases among microorganisms has been undertaken. Sixteen hexose kinases of five major types were partially purified from 12 microorganisms and characterized with respect to specificity for sugar and nucleotide substrates and Michaelis constants for the sugar substrates. Glucokinase activities that phosphorylate glucose and glucosamine are inhibited by N-acetyl-glucosamine and xylose, were found to be present in the non-sulphur photosynthetic bacteria Rhodospirillum rubrum, the blue-green algae Anacystis montana, and the protists Chlorella pyrenoidosa and Chlamydomonas reinhardtii (green algae), Hypochytrium catenoides (Hypochytridiomycete) and Saprolegnia Iitoralis (Oomycete). The myxobacteria Stigmatella aurantiaca contains a glucokinase activity with a different specificity pattern. Anacystis and Chlorella, besides their glucokinase activities, contain highly specific fructokinases, although that from Anacystis can also phosphorylate fructosamine; fructokinase from Anacystis has a molecular weight of 20 000, and exhibits a sigmoidal saturation curve for ATP when the Mg2+/ATP ratio is 2; this curve is transformed to a Michaelian one when under the same conditions an excess of Mg2+ (5 mM) is added. Saprolegnia however, besides the glucokinase, contains a mannofructokinase activity that phosphorylates mannose (Km 0.06 mM) and fructose (1 mM). On the other hand, hexokinase, a low specificity enzyme, was detected in the protist Allomyces arbuscula (Chytridiomycete) and in fungi Mucor hiemalis and Phycomyces blakesleeanus (Zygomycetes), and Schizophyllum commune (Basidiomycete). Schizophyllum contains a glucomannokinase activity together with hexokinase activity. The pattern of distribution of ATP-dependent hexose kinases among microorganisms seems to parallel that reported for biosynthetic pathways for lysine. The correlation with other biochemical parameters is also considered.

Bacteria↗

Microbial production of hydroxy and oxo fatty acids by several microorganisms as a model of adipocere formation.

Some varieties of aerobic or anaerobic microorganisms from the human stool and adipocere were separated and identified. These separated microorganisms together with other authentic ones produced 10-hydroxystearic acid from oleic acid. Some bacteria could convert oleic acid to 10-oxostearic acid as well as 10-hydroxystearic acid. These findings indicate that the microbial enzyme(s) catalyzes the hydration of oleic acid and probably the oxidation of this hydrated product. Aerobic bacteria as well as anaerobic microorganisms were found to be involved in the formation of adipocere.

Bacillus subtilis↗

Harmonic 'signatures' of microorganisms.

The frequency/amplitude effect of various microorganisms exposed to periodic (time varying) electric fields, when proximate to immersed electrodes, has been studied using a novel analytical instrument. The harmonic distribution, in complex signals caused by cells exposed to harmonic free waveforms and occupying part of the electrode/suspension interface volume, was shown to be almost entirely due to the change in the standing interfacial transfer function by the (dielectrically nonlinear) presence of cells. Thus, the characteristic interfacial non-linearity is viewed as variable, being uniquely modulated by the presence of particular cells in the interfacial region. Little can be attributed to bulk (far field) effects. The tendency for subtle (characteristic) signal distortion to occur as a function of particulate (cell or molecular) occupancy of the near electrode interfacial region under controlled current conditions leads to the method of sample characterisation by harmonic (Fourier) analysis. We report here, as a sequel to our original studies (Hutchings et al., 1993; Hutchings and Blake-Coleman, 1993), preliminary results of the harmonic analysis of microbial suspensions under controlled signal conditions using a three-electrode configuration. These data provide three-dimensional graphical representations producing harmonic 'surfaces' for various microorganisms. Thus, cell type differences are characterised by their 'harmonic signature'. The visual distinction provided by these 'surface' forming three-dimensional plots is striking and gives a convincing impression of the ability to identify and enumerate specific microorganisms by acquisition of cell-modulated electrode interfacial Fourier spectra.

Candida↗

Surface transport of microorganisms by water.

Several studies have reported on the quality of runoff from land that has received either an application of livestock waste or been utilized as a pasture for livestock. Unfortunately, these studies have not directed their efforts to understanding and developing the relationships among several of the important parameters that influence runoff quality. One of the reasons for this deficiency is that the list of influencing parameters is quite long. Nevertheless, it is important to identify the parameters and their probable impact on movement of organisms in water. The microbiological aspects are influenced by the fate of organisms in the environment. Radiant energy (sunlight), temperature, available nutrients, presence of toxic materials, available moisture (precipitation and humidity), and soil pH all influence the death/growth rate of the organisms in question. Site characteristics, such as slope, vegetative cover, antecedent moisture content, soil type, organic matter content, infiltration rate, and surface condition of the soil, all influence microorganism movement. Hydrologic factors, such as frequency, duration, and intensity of rainfall, are very critical in determining the characteristics of runoff events that provide the transportation to move introduced organisms from their application site. There are very few models today that can be used to calculate the microorganism population in runoff. While many of the influencing parameters have been identified, there has been little research on the surface transport of microorganisms.

Animals↗

Fatty acid biosynthesis in microorganisms being used for Single Cell Oil production.

Single cell oils (SCOs) are now produced by various microorganisms as commercial sources of arachidonic acid (ARA) and docosahexaenoic acid (DHA). These oils are now used extensively as dietary supplements in infant formulas. An understanding of the underlying biochemistry and genetics of oil accumulation in such microorganisms is therefore essential if lipid yields are to be improved. Also an understanding of the biosynthetic pathways involved in the production of these polyunsaturated fatty acids (PUFAs) is also highly desirable as a prerequisite to increasing their content in the oils. An account is provided of the biosynthetic machinery that is necessary to achieve oil accumulation in an oleaginous species where it can account for lipid build up in excess of 70% of the cell biomass. Whilst PUFA production in most microorganisms uses a conventional fatty acid synthase (FAS) system followed by a series of desaturases and elongases, in Schizochytrium sp., and probably related thraustochytrid marine protists, PUFA synthesis now appears to be via a polyketide synthase (PKS) route. This route is discussed. It clearly represents a major departure from conventional fatty acid biosynthesis, possibly as a means of decreasing the amount of NADPH that is needed in the overall process.

Acetyltransferases↗

Compost microbiomes as reservoirs of cellulolytic microorganisms for cellulosic textile degradation.

Cellulosic textiles, constituting over 30% of global fibre production, are biodegradable but remain challenging to recycle at scale owing to their high crystallinity, chemical finishes, and heterogeneous waste streams. Although microorganisms drive cellulose turnover in natural ecosystems, their potential for transforming anthropogenic cellulosic waste remains largely unexplored. In this study, composting was evaluated both as a sustainable approach to textile biodegradation and a reservoir of cellulolytic microorganisms with biotechnological potential. Biodegradation assays of cotton and lyocell were integrated with shotgun metagenomics and targeted cultivation to identify microbial taxa and enzymes involved in cellulose degradation. Composting trials showed that degradation was strongly influenced by both composting system and fibre composition. Community composting achieved near-complete textile disintegration, while shredded textiles exhibited the highest degradation rates, reaching up to 97%. Shotgun metagenomic revealed a bacterial-dominated community enriched in Actinomycetota and Bacillota and characterised by an abundance of glycoside hydrolases. Culture-based screening recovered 62 microbial isolates, of which Neurospora and Aspergillus exhibited the highest cellulolytic activity (>60%). In vitro assays further showed that cotton was more readily degraded than lyocell, with several isolates achieving >70% mass loss. Metagenomic approach revealed a predominantly bacterial composting community at the sampled stage, whereas cultivation preferentially recovered fungi that, despite their low relative abundance in situ, exhibited strong cellulolytic potential. These findings highlight the potential of composting as a sustainable end-of-life strategy for cellulosic textiles and identify compost microbiomes as valuable reservoirs of cellulolytic microorganisms for the development of sustainable bioprocesses for textile waste treatment.

Cellulose↗

Genomics, metagenomics and proteomics in biomining microorganisms.

The use of acidophilic, chemolithotrophic microorganisms capable of oxidizing iron and sulfur in industrial processes to recover metals from minerals containing copper, gold and uranium is a well established biotechnology with distinctive advantages over traditional mining. A consortium of different microorganisms participates in the oxidative reactions resulting in the extraction of dissolved metal values from ores. Considerable effort has been spent in the last years to understand the biochemistry of iron and sulfur compounds oxidation, bacteria-mineral interactions (chemotaxis, quorum sensing, adhesion, biofilm formation) and several adaptive responses allowing the microorganisms to survive in a bioleaching environment. All of these are considered key phenomena for understanding the process of biomining. The use of genomics, metagenomics and high throughput proteomics to study the global regulatory responses that the biomining community uses to adapt to their changing environment is just beginning to emerge in the last years. These powerful approaches are reviewed here since they offer the possibility of exciting new findings that will allow analyzing the community as a microbial system, determining the extent to which each of the individual participants contributes to the process, how they evolve in time to keep the conglomerate healthy and therefore efficient during the entire process of bioleaching.

Bacteria↗

Biotechnological approaches for the production of polyhydroxyalkanoates in microorganisms and plants - a review.

The increasing effect of non-degradable plastic wastes is a growing concern. Polyhydroxyalkanoates (PHAs), macromolecule-polyesters naturally produced by many species of microorganisms, are being considered as a replacement for conventional plastics. Unlike petroleum-derived plastics that take several decades to degrade, PHAs can be completely bio-degraded within a year by a variety of microorganisms. This biodegradation results in carbon dioxide and water, which return to the environment. Attempts based on various methods have been undertaken for mass production of PHAs. Promising strategies involve genetic engineering of microorganisms and plants to introduce production pathways. This challenge requires the expression of several genes along with optimization of PHA synthesis in the host. Although excellent progress has been made in recombinant hosts, the barriers to obtaining high quantities of PHA at low cost still remain to be solved. The commercially viable production of PHA in crops, however, appears to be a realistic goal for the future.

Alkanes↗

Mechanism by which Bombyx mori hemocytes recognize microorganisms: direct and indirect recognition systems for PAMPs.

Hemocytes play an important role in cellular reactions in the immune system. Although the recognition of pathogens is thought to involve pattern-recognition proteins (PRPs) in insects, the exact mechanisms by which insect hemocytes recognize pathogens are not clear. This study examined the mechanism by which Bombyx mori hemocytes recognize microorganisms and pathogen-associated molecular patterns (PAMPs) using flow cytometry and fluorescence microscopy. Fluorescence-labeled bacterial or fungal cells were observed to bind to hemocytes and this binding was inhibited by adding lipoteichoic acid (LTA) or beta-1,3-glucan. Lipopolysaccharide (LPS) bound to hemocytes directly. These results suggest that hemocytes have a mechanism that recognizes LPS, LTA, and beta-1,3-glucan directly. Previously, we identified two types of C-type lectin (BmLBP and BmMBP) and showed that they recognize a variety of PAMPs leading to the induction of nodule formation. These lectins enhanced hemocyte binding to microorganisms and their direct binding to hemocytes suggests that hemocytes have a mechanism for recognizing microorganisms using lectin receptors.

Animals↗

Removal of triazine herbicides from freshwater systems using photosynthetic microorganisms.

The uptake of the triazine herbicides, atrazine and terbutryn, was determined for two freshwater photosynthetic microorganisms, the green microalga Chlorella vulgaris and the cyanobacterium Synechococcus elongatus. An extremely rapid uptake of both pesticides was recorded, although uptake rate was lower for the cyanobacterium, mainly for atrazine. Other parameters related to the herbicide bioconcentration capacity of these microorganisms were also studied. Growth rate, biomass, and cell viability in cultures containing herbicide were clearly affected by herbicide uptake. Herbicide toxicity and microalgae sensitivity were used to determine the effectiveness of the bioconcentration process and the stability of herbicide removal. C. vulgaris showed higher bioconcentration capability for these two triazine herbicides than S. elongatus, especially with regard to terbutryn. This study supports the usefulness of such microorganisms, as a bioremediation technique in freshwater systems polluted with triazine herbicides.

Atrazine↗

Water activity affects heat resistance of microorganisms in food powders.

To study the factors and mechanisms involved in microorganisms' death or resistance to temperature in low-water-activity environments, a previous work dealt with the viability of dried microorganisms immobilized in thin-layer on glass beads. This work is intended to check the efficiency of a rapid heating-cooling treatment to destroy microorganisms that were dried after mixing with wheat flour or skim milk. The thermoresistance of the yeast Saccharomyces cerevisiae and the bacterium Lactobacillus plantarum were studied. Heat stress was applied at two temperatures (150 or 200 degrees C) for treatments of one of four durations (5, 10, 20, or 30 s) and at seven levels of initial water activity (a(w)) in the range 0.10 to 0.70. This new treatment achieved a microbial destruction of eight log reductions. A specific initial water activity was defined for each strain at which it was most resistant to heat treatments. On wheat flour, this initial a(w) value was in the range 0.30-0.50, with maximal viability value at a(w)=0.35 for L. plantarum, whatever the temperature studied, and 0.40 for S. cerevisiae. For skim milk, a variation in microbial viability was observed, with optimal resistance in the range 0.30-0.50 for S. cerevisiae and 0.20-0.50 for L. plantarum, with minimal destruction at a(w)=0.30 whatever the heating temperature is.

Food Contamination↗