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Dissimilation of tryptophan and related indolic compounds by ruminal microorganisms in vitro.

Intraruminal doses of L-tryptophan cause acute pulmonary edema and emphysema in cattle. The D and L isomers of tryptophan and 22 related indolic compounds were incubated with ruminal microorganisms in vitro. Incubation of L-[U-benzene ring-(14)C]tryptophan with ruminal microorganisms for 24 h resulted in 39% of the added radioactivity being incorporated into skatole, 7% into indole, and 4% into indoleacetate (IAA). D-Tryptophan was not degraded to any of these metabolites. The major pathway of skatole formation from L-tryptophan appeared to be by the decarboxylation of IAA. Incubation of [2-(14)C]IAA with ruminal microorganisms for 24 h resulted in 38% incorporation into skatole. L-[5-Hydroxy]tryptophan was degraded to 5-hydroxyskatole and 5-hydroxyindole, whereas 5-hydroxyindoleacetate was degraded to only 5-hydroxyskatole. Incubation of indolepyruvate, indolelactate, and indolealdehyde with ruminal microorganisms resulted in the formation of both skatole and indole. Under similar conditions, indoleacetaldehyde was converted to IAA and tryptophol. The addition of increasing concentrations of glucose (0 to 110 mM) reduced the formation of both skatole and indole from L-tryptophan and resulted in the accumulation of IAA. Antibiotics reduced the degradation of L-tryptophan to skatole and indole, with kanamycin and neomycin particularly effective in reducing the decarboxylation of IAA to skatole.

5-Hydroxytryptophan↗

Extraction and partial characterization of a leukotoxin from a plaque-derived Gram-negative microorganism.

The plaque-derived gram-negative microorganism Y4 identified as a member of the genus Actinobacillus, was tested for a soluble cytotoxic factor(s). Sonication or incubation of viable Y4 microorganisms in distilled water or normal human serum resulted in liberation of a soluble material which was cytotoxic in vitro for human polymorphonuclear leukocytes (PMNs). The Y4 soluble sonic extract was also cytotoxic to human peripheral blood monocytes. However, human lymphocytes, platelets, and fibroblasts, as well as rabbit, rat, and mouse leukocytes and chicken embryo fibroblasts, were not killed by exposure to the Y4 sonic extract. No hemolytic activity was detected in the Y4 sonic extract. No hemolytic activity was detected in the Y4 sonic extract. Consequently, the factor(s) in the Y4 sonic extract was referred to as Y4 leukotoxin. The Y4 leukotoxin was inactive at 4 degrees C, heat sensitive (56 degrees C, 30 min), and inactivated by proteases. The cytotoxic effect of Y4 leukotoxin on PMNs was dose, time, and temperature dependent. The leukotoxin did not bind to viable PMNs at 4 degrees C but did bind to dead PMN membrane components at both 4 and 37 degrees C. The addition of bovine serum albumin (51 mg/ml) to PMN-Y4 leukotoxin cultures inhibited the release of lactate dehydrogenase from the PMNs, but did not prevent the death of the cells as indicated by electron microscopy. Lysosomal markers were released in parallel to the cytoplasmic enzyme lactate dehydrogenase from Y4 leukotoxin-treated PMNs. The addition of 0.02 M ethylenedinitrilotetraacetic acid to these cultures inhibited release of lysosomal markers but enhanced the release of lactate dehydrogenase. These results suggested that a soluble leukotoxin with specificity for only human PMNs and monocytes can be liberated from viable Y4. What role this leukotoxin plays in the pathogenicity of the Y4 microorganism is not yet known. However, this leukotoxin is one of the first materials from a plaque-derived microorganism with a potential role in the pathogenesis of juvenile periodontitis.

Actinobacillus↗

Effect of atypical antibiotic resistance on microorganism identification by pattern recognition.

We classified microorganisms from the clinical laboratory by using information provided by the Gram stain and antibiotic sensitivity profiles obtained with the Bauer-Kirby technique. Approximately 4,000 microorganisms, routinely identified and tested for antibiotic sensitivities in a large hospital microbiology laboratory, were used as a data set for several pattern recognition classification methods: K--nearest-neighbor analysis, statistical isolinear multicomponent analysis, Bayesian inference, and linear discriminant analysis. K--nearest-neighbor analysis yielded the highest prospective classification accuracy for gram-negative organisms, 90%. When those organisms displaying an atypical antibiotic resistance pattern were excluded from the data, the gram-negative classification accuracy improved to 95%. These results are inferior to currently accepted biochemical identification methods. Microorganisms with atypical antibiotic resistance patterns are likely to be misidentified and are common enough (17% of our isolates) to limit the feasibility of routine identification of microorganisms from their antibiotic sensitivities.

Anti-Bacterial Agents↗

Metagenomics: application of genomics to uncultured microorganisms.

Metagenomics (also referred to as environmental and community genomics) is the genomic analysis of microorganisms by direct extraction and cloning of DNA from an assemblage of microorganisms. The development of metagenomics stemmed from the ineluctable evidence that as-yet-uncultured microorganisms represent the vast majority of organisms in most environments on earth. This evidence was derived from analyses of 16S rRNA gene sequences amplified directly from the environment, an approach that avoided the bias imposed by culturing and led to the discovery of vast new lineages of microbial life. Although the portrait of the microbial world was revolutionized by analysis of 16S rRNA genes, such studies yielded only a phylogenetic description of community membership, providing little insight into the genetics, physiology, and biochemistry of the members. Metagenomics provides a second tier of technical innovation that facilitates study of the physiology and ecology of environmental microorganisms. Novel genes and gene products discovered through metagenomics include the first bacteriorhodopsin of bacterial origin; novel small molecules with antimicrobial activity; and new members of families of known proteins, such as an Na(+)(Li(+))/H(+) antiporter, RecA, DNA polymerase, and antibiotic resistance determinants. Reassembly of multiple genomes has provided insight into energy and nutrient cycling within the community, genome structure, gene function, population genetics and microheterogeneity, and lateral gene transfer among members of an uncultured community. The application of metagenomic sequence information will facilitate the design of better culturing strategies to link genomic analysis with pure culture studies.

Biotechnology↗

Influence of aerobic microorganisms upon virus survival in soil.

Survival of human poliovirus type 1 in a sandy loam soil appeared to be deleteriously influenced by aerobic microorganisms. This effect was determined by comparing the survival of virus in soil under four different possible combinations of aerobic versus anaerobic (H2-CO2) atmosphere and sterile versus nonsterile condition. Storage of samples was done in humid chambers to prevent soil desiccation. The effect attributed to aerobic microorganisms was measurable and statistically significant at all three incubation temperatures used in the study (1, 23, and 37 degrees C), with the increase in inactivation rate attributable to aerobic microorganisms generally being two to threefold. No comparable effect was observed to occur for anaerobic microorganisms under the sets of conditions employed in the study.

Aerobiosis↗

Survey, purification, and properties of sugar phosphate phosphohydrolase among microorganisms.

Sugar phosphate phosphohydrolase was purified approximately 500- to 600-fold to apparent homogeneity from Escherichia coli B, Escherichia coli C, Escherichia coli var. communior, Escherichia acidilactici, Enterobacter aerogenes, Neisseria meningitidis, and Saccharomyces cereviseae. The molecular weights of the enzyme as estimated by gel filtration ranged from 97 X 10(3) to 101 X 10(3). The enzyme was composed of two subunits with the same molecular weight which ranged from 50 X 10(3) to 52 X 10(3), as determined by sodium dodecyl sulfate gel electrophoresis. Homogeneous enzyme preparations hydrolyse all the tested alpha-D-aldohexose 1-phosphate, D-(keto or aldo)hexose 6-phosphate, and pentose phosphate substrates significantly. When the microorganisms were transferred from growth medium with 1% glucose to that without glucose, there were dramatic increases in both the specific and total enzyme activities. At least three isozymes appeared to be present in S. cereviseae, and two appeared to be present in E. coli B, E. coli var. communior, and N. meningitidis. Rabbit antiserum immunized against sugar phosphate phosphohydrolase purified from E. coli B cross-reacted with both the crude extracts and purified preparations of the enzyme from the other microorganisms. The presence of neither sugar phosphate phosphohydrolase activity nor immunocrossreacting material was detected in the following microorganisms: Aspergillus niger, Azotobacter chroococcum, Bacillus subtilis, Bacillus pumilis, Citrobacter freundii, Clostridium butyricum, Corynebacterium xerosis, Flavobacterium aquatile, Flavobacterium synxanthum, Lactobacillus bulgaricus, Micrococcus coralinus, Neisseria perflava, Neurospora crassa, Penicilium expansum, Penicilium notatum, Proteus mirabilis, Proteus vulgaris, Pseudomonas fluorescens, Saccharomyces fermenti, Sarcina lutea, and Streptomyces antibioticus. At present, no conclusive relationship can be established between the phosphoenolpyruvate phosphotransferase system and the enzyme sugar phosphate phosphohydrolase among microorganisms. The physiological role of sugar phosphate phosphohydrolase as a transferase and regulatory enzyme is discussed.

Bacteria↗

Microorganism transport in the human endolymphatic duct.

There are indications that endolymphatic sac (ES) may be an immunologically active part of the inner ear. So far, no microorganisms or foreign substances have been localized in this area under 'normal' conditions. Only a limited number of human specimens, including the entire endolymphatic duct (ED) and ES, have been collected and analyzed from cadavers or surgical biopsy specimens. In this study, 6 human ED and ES collected from cadavers and at surgery were analyzed by light and electron microscopy. This was done in order to investigate if microorganisms may normally be drained at this route into the ES. Some microorganisms (Mycoplasma pneumoniae) were found in the lumen and subepithelial tissue of 1 human ED. These observations suggest that microorganisms may also be locally processed and disposed at the level of the ED. These results add further evidence as to the immunodefensive role of the human ES.

Endolymphatic Duct↗

Oropharyngeal flora in asthma and in chronic obstructive pulmonary disease. Indigenous oropharyngeal microorganisms in outpatients with asthma or chronic obstructive pulmonary disease.

In vitro and in vivo studies have shown that various strains of "viridans streptococci" (nongroupable alpha-hemolytic streptococci) inhabiting the oropharynx suppress the growth of gram-positive and gram-negative microorganisms. We conducted an inventory of the oropharyngeal flora from ambulatory asthma and chronic obstructive pulmonary disease (COPD) patients and a control group to examine the interaction between viridans streptococci and potential pathogens in vivo. In addition, the difference in colonization patterns of these bacteria was studied. Oral washings from 195 patients, 48 asthma (24.6%), 147 COPD (75.4%), and 157 control subjects were examined microbiologically on two occasions with a 2-wk interval, resulting in a total of 384 and 295 oral washings, respectively. All patients were in a stable phase of disease throughout the study. The distribution of low (< or = 10(4)/ml) or high (> or = 10(5)/ml) concentrations of viridans streptococci did not differ substantially between asthma or COPD patients and control subjects. Potentially pathogenic microorganisms found in a low (< or = 10(4)/ml) or high (> or = 10(5)/ml) concentration were equally distributed between the two groups. Staphylococcus aureus and beta-hemolytic streptococci were found significantly less often in the asthma and COPD group (p < 0.005 and p < 0.0005, respectively), but the prevalence of Enterobacteriaceae species was significantly higher (p < 0.0005). No correlation was found between the concentration of viridans streptococci and the prevalence of gram-negative microorganisms. These findings suggest that viridans streptococci are probably not responsible for growth control of gram-negative microorganisms in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Surface germicidal effects of ozone for microorganisms.

In this study the influences of microorganism species, relative humidity, and ozone dosage on ozone surface disinfection were evaluated. Bacterial and fungal cultures were spread on agar plates and exposed to ozone. The selected microorganisms included Escherichia coli, Bacillus subtilis, Candida famata, and Penicillium citrinum. Results showed that microorganism survival fraction and ozone dosage (ozone concentration times exposure time) have an exponential relationship. Results also indicated that E. coli was the most sensitive organism to ozone exposure. E. coli required only very low ozone doses of 2-2.5 and 3.5-4 mg to obtain 50 and 80% inactivation, respectively. In addition, P. citrinum was more resistant than E. coli and required ozone doses of 40-60 and 60-120 mg to obtain 50 and 80% inactivation. In addition, spores of B. subtilis were observed to be the most resistant organism, requiring ozone doses of 40-75 and 145-150 mg to obtain 50 and 80% inactivation. Yeast was less resistant than P. citrinum and B. subtilis, requiring ozone doses of 10 and 15-19 mg to obtain 50 and 80% inactivation. It was clearly indicated that the ozone dose differences for 80% microorganism inactivation could be as high as 40 times between B. subtilis and E. coli. Ozone surface germicidal efficiency increased as relative humidity increased, which could be related to more radicals generated from ozone reaction with more water vapor at higher relative humidity. It was concluded that ozone should be highly effective and provide a reliable safety factor in treating contaminated surface. In addition, workers might need to wear suitable respiratory protection at high ozone level operation.

Bacteria↗

Opsonic activity of cord blood sera against various species of microorganism.

The results of the present study on the opsonic activity of cord blood serum against various microorganisms (Staphylococcus aureus, Escherichia coli, and group B streptococci) show that the activity of cord blood serum in promoting IgG-mediated ingestion is equal to that of sera of healthy adults. This implies that IgG concentrations, as assessed by immunochemical methods, in cord blood and adult sera represent functionally similar IgG activities. Ingestion of microorganisms involving complement-dependent opsonization was found to be of the same level for cord blood and adult sera, when the opsonization occurred via the classical pathway of complement activation. However, due to decreased concentrations of factors B, P, and D in cord blood serum, optimal opsonization of microorganisms requiring the alternative pathway of complement was impaired. Taken together, these results indicate that an opsonic defect of cord blood serum affects mainly microorganisms requiring opsonization via the alternative pathway of complement.

Blood Bactericidal Activity↗

Prevalence of microorganisms in root canals of human deciduous teeth with necrotic pulp and chronic periapical lesions.

The objective of this study was to evaluate bacterial prevalence in 31 root canals of human deciduous teeth with necrotic pulp and periapical lesions using bacterial culture. After crown access, the material was collected using absorbent paper points for microbiological evaluation and determination of colony forming units (CFU). Anaerobic microorganisms were found in 96.7% of the samples, black-pigmented bacilli in 35.5%, aerobic microorganisms in 93.5%, streptococci in 96.7%, and S. mutans in 48.4%. We concluded that in human deciduous teeth root canals with necrotic pulp and periapical lesions the infection is polymicrobial, with a large number of microorganisms and a predominance of streptococci and anaerobic microorganisms.

Child↗

Structure of dried cellular alginate matrix containing fillers provides extra protection for microorganisms against UVC radiation.

Soil microorganisms in general and biocontrol agents in particular are very sensitive to UV light. The packaging of biocontrol microorganisms into cellular solids has been developed as a means of reducing loss caused by exposure to environmental UV radiation. The bacterial and fungal biocontrol agents Pantoea agglomerans and Trichoderma harzianum were immobilized in freeze-dried alginate beads containing fillers and subjected to 254 nm UV radiation (UVC). Immobilization of cells in freeze-dried alginate-glycerol beads resulted in greater survival after UV irradiation than for a free cell suspension. Adding chitin, bentonite or kaolin as fillers to the alginate-glycerol formulation significantly increased bacterial survival. Immobilization in alginate-glycerol-kaolin beads resulted in the highest levels of survival. The transmissive properties of the dried hydrocolloid cellular solid had a major influence on the amount of protection by the cell carrier. Dried alginate matrix (control) transmitted an average of 7.2% of the radiation. Filler incorporation into the matrix significantly reduced UV transmission: Alginate with kaolin, bentonite and chitin transmitted an average of 0.15, 0.38 and 3.4% of the radiation, respectively. In addition, the filler inclusion had a considerable effect on the bead's average wall thickness, resulting in a approximately 1.5- to threefold increase relative to beads based solely on alginate. These results suggest that the degree of protection of entrapped microorganisms against UVC radiation is determined by the UV-transmission properties of the dried matrix and the cellular solid's structure. It is concluded that for maximum protection against UV-radiation-induced cell loss, biocontrol microorganisms should be immobilized in alginate-glycerol beads containing kaolin.

Alginates↗

The analysis of microorganisms by microcalorimetry in the pharmaceutical industry.

Many features of microorganisms make them pre-eminently suitable for study by microcalorimetry. They have thus, in the past, been the basis of fundamental studies in metabolism and cellular physiology. In this review we look at the application of calorimetry to the impact of bacteria and fungi on the pharmaceutical industry both in the exploitation of useful microorganisms and the fight against harmful ones. Obviously they are of great relevance to the pharmaceutical industry as agents of human disease, with more antimicrobial products registered for production than for any other type of human affliction. Microcalorimetry offers the opportunity to study microorganisms in real time and in heterogeneous systems, allowing for more descriptive and representative analysis. Other advantages that microcalorimetry confers over traditional microbiological techniques are reductions in time, better reproducibility and simplicity. Also the manufacture of all pharmaceutical products requires the exclusion of microorganisms to a greater or lesser degree. The enumeration and identification of such contaminants is of great importance for the well-being of patients and to maintain the integrity of the product. New techniques are required to increase the reliability and sensitivity over conventional methods. Finally, as our understanding of biology develops, the sophistication of therapeutic agents available, such as vaccines, cytokines and engineered antibodies, is increasing. Necessarily, prokaryotic and eukaryotic cells, possibly transformed with the appropriate genes, are the producers of such proteins. Microcalorimetry offers a sensitive means of developing the conditions for optimum production of such products in active form since it gives instantaneous information on the physiology of the producer cell.

Bacteria↗

[Survival conditions of microorganisms under extremely severe environment].

The survival conditions of microorganisms under extremely severe environment are of interest in various areas of biology, sterilization, and space engineering, especially where resistance to microorganisms is concerned. Despite the interest, the resistance to microorganisms under extremely severe environment such as space environment or other planetary environment is not known well. In order to investigate survival conditions of microorganisms under extremely severe environment, surviving fractions for spores and vegetative cells of Bacillus subtilis were surveyed in various chemical species of atmosphere at various pressures and various temperatures, and the dependence on time for surviving fractions was examined. The results show: (i) Surviving fractions depend on chemical species of atmosphere. (ii) At high pressure and high temperature, surviving fractions are low and the resistance of spores is stronger than that of vegetative cells. (iii) Surviving fractions decrease as first-order reaction along with time elapsed.

Argon↗

[Microbial diversity of deep-sea extremophiles--Piezophiles, Hyperthermophiles, and subsurface microorganisms].

Knowledge of our Planet's biosphere has increased tremendously during the last 10 to 20 years. In the field of Microbiology in particular, scientists have discovered novel "extremophiles", microorganisms capable of living in extreme environments such as highly acidic or alkaline conditions, at high salt concentration, with no oxygen, extreme temperatures (as low as -20 degrees C and as high as 300 degrees C), at high concentrations of heavy metals and in high pressure environments such as the deep-sea. It is apparent that microorganisms can exist in any extreme environment of the Earth, yet already scientists have started to look for life on other planets; the so-called "Exobiology" project. But as yet we have little knowledge of the deep-sea and subsurface biosphere of our own planet. We believe that we should elucidate the Biodiversity of Earth more thoroughly before exploring life on other planets, and these attempts would provide deeper insight into clarifying the existence of extraterrestrial life. We focused on two deep-sea extremophiles in this article; one is "Piezophiles", and another is "Hyperthermophiles". Piezophiles are typical microorganisms adapted to high-pressure and cold temperature environments, and located in deep-sea bottom. Otherwise, hyperthermophiles are living in high temperature environment, and located at around the hydrothermal vent systems in deep-sea. They are not typical deep-sea microorganisms, but they can grow well at high-pressure condition, just like piezophiles. Deming and Baross mentioned that most of the hyperthermophilic archaea isolated from deep-sea hydrothermal vents are able to grow under conditions of high temperature and pressure, and in most cases their optimal pressure for growth was greater than the environmental pressure they were isolated from. It is possible that originally their native environment may have been deeper than the sea floor and that there had to be a deeper biosphere. This implication suggests that the deep-sea hydrothermal vents are the windows to a deep subsurface biosphere. A vast array of chemoautotrophic deep-sea animal communities have been found to exist in cold seep environments, and most of these animals are common with those found in hydrothermal vent environments. Thus, it is possible to consider that the cold seeps are also one of slit windows to a deep subsurface biosphere. We conclude that the deep-sea extremophiles are very closely related into the unseen majority in subsurface biosphere, and the subsurface biosphere probably concerns to consider the "exobiology".

Archaea↗

Infection of colonized cat fleas, Ctenocephalides felis (Bouché), with a rickettsia-like microorganism.

We report the ultrastructure of a rickettsia-like microorganism in a colonized population of the cat flea Ctenocephalides felis (Bouché). The microorganism occurs principally in the cytoplasm of midgut cells, but similar microorganisms were detected in the tracheal matrix, muscle, hypodermis, ovaries, and the epithelial sheath of the testes. The microorganism has a well-defined cell membrane consistent with rickettsia and measures 0.25-0.45 microns in diameter with lengths up to 1.5 microns. It was observed repeatedly in fleas of 1 laboratory colony, including newly emerged non-bloodfed specimens, but not in specimens from several other sources.

Alphaproteobacteria↗

An annotated checklist of pathogenic microorganisms associated with migratory birds.

The potential for transport and dissemination of certain pathogenic microorganisms by migratory birds is of concern. Migratory birds might be involved in dispersal of microorganisms as their biological carriers, mechanical carriers, or as carriers of infected hematophagous ecto-parasites (e.g., ixodid ticks). Many species of microorganisms pathogenic to homeothermic vertebrates including humans have been associated with free-living migratory birds. Migratory birds of diverse species can play significant roles in the ecology and circulation of some arboviruses (e.g., eastern and western equine encephalomyelitis and Sindbis alphaviruses, West Nile and St. Louis encephalitis flaviviruses), influenza A virus, Newcastle disease virus, duck plague herpes-virus, Chlamydophila psittaci, Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Campylobacter jejuni, Salmonella enterica, Pasteurella multocida, Mycobacterium avium, Candida spp., and avian hematozoans. The efficiency of dispersal of pathogenic microorganisms depends on a wide variety of biotic and abiotic factors affecting the survival of the agent in, or disappearance from, a habitat or ecosystem in a new geographic area.

Animal Migration↗

Deoxyribonucleic acid typing methods for medically important microorganisms.

The ability to type microorganisms to a sub-species level plays an essential role in the diagnosis, treatment and control of human infection. Traditionally, differentiation of microorganisms has involved analysis of phenotypic markers. However, these methods are not universally applicable to all microorganisms, and results may be influenced by environmental factors. Recent developments in DNA analysis, together with the limitations of phenotypic methods, have resulted in an increasing use of procedures based on DNA analysis for the typing of clinically important microorganisms. The aim of this review is to provide an overview of the advantages and disadvantages of the genetic typing techniques currently available.

Bacterial Typing Techniques↗