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PubMed2005

Real-time quantitative PCR with gene probe, fluorochrome and flow cytometry for microorganism analysis.

Microorganism concentrations and viability can be better understood and clarified by using both culture and non-culture methods. Here, using pure suspensions of E. coli, three non-culture methods, namely, flow cytometry (FCM), epifluorescence microscopy (EFM), and real-time quantitative polymerase chain reaction (real-time qPCR), were compared with a traditional culture-based method. Using fluorocome-labeling methods with FCM and EFM applications, acridine orange (AO) and propidium iodide (PI) dyes were used to determine the total cell concentration and microorganism viability, respectively. The results indicated that total cell concentrations determined using FCM were statistically higher (2.62-4.94 times) than those determined using EFM. The difference might be due to cell losses induced by extensive preparations needed for EFM. In addition, EFM and FCM were highly associated for both the total cell concentration and viability. FCM-measured viability was the highest, whereas the culture-measured viability was the lowest. Furthermore, DNA concentrations measured by real-time qPCR with gene probe were highly associated with the total number concentrations measured by either the EFM or FCM. In summary, the three non-culture methods compared here could provide rapid and accurate information about microorganism concentrations and viabilities.

Acridine Orange↗
PubMed2003

Microorganisms from canals of root-filled teeth with periapical lesions.

AIM: The objective of the present study was to identify the microbial flora within root canals of teeth with failed root-canal treatment and to determine the association of the various species with clinical features. METHODOLOGY: Sixty root-filled teeth with persisting periapical lesions were selected for this study. During nonsurgical endodontic re-treatment, the root-filling material was removed and the canals were sampled. Microbial sampling, isolation and species determination were performed using advanced microbiological techniques for anaerobic species. The association of microbiological findings with clinical features was investigated. RESULTS: Microorganisms were recovered from 51 teeth. In most cases, one or two strains per canal were found. Of the microbial species isolated, 57.4% were facultative anaerobic species and 83.3% Gram-positive microorganisms. Enterococcus faecalis was the most frequently recovered bacterial species. Obligate anaerobes accounted for 42.6% of the species and the most frequently isolated genera was Peptostreptococcus. which was associated with clinical symptoms (P < 0.01). Significant associations were also observed between: (a) pain or history of pain and polymicrobial infections or anaerobes (P < 0.05): (b) tenderness to percussion and Prevotella intermedia/P. nigrescens (P < 0.05); (c) sinus and Streptococcus spp. (P < 0.001) or Actinomyces spp. (P < 0.01); (d) coronally unsealed teeth and Streptococcus spp. or Candida spp. (both with P < 0.01). CONCLUSION: The microbial flora in canals after failure of root-canal treatment were limited to a small number of predominantly Gram-positive microbial species. Facultative anaerobes, especially E. faecalis, were the most commonly isolated microorganisms, however, polymicrobial infections and obligate anaerobes were frequently found in canals of symptomatic root-filled teeth.

Bacteria, Anaerobic↗
PubMed1997

Existence of Candida albicans and microorganisms in denture stomatitis patients.

The aetiology of denture stomatitis is not clear from the literature. Some studies show its aetiology as Candida albicans, while other reports point out the significance of microorganisms. In this study the existence of C. albicans and microorganisms was investigated in subjects with and without denture stomatitis. The results showed that a combination of C. albicans and microorganisms is more likely to be responsible for denture stomatitis.

Bacteria↗
PubMed1996

Photobiology of microorganisms: how photosensors catch a photon to initialize signalling.

Photobiological processes are relevant for microorganisms for energy generation, protection against excess and/or damaging radiation, and for signalling. In this review we give an overview of the knowledge on the functioning of photosensors in microorganisms, with special emphasis on the conformational changes that lead to signal generation and transduction. Light is absorbed by specific chromophores, which are tuned, by their proteinaceous environment, to function optimally. These chromophores belong to three classes: tetrapyrroles, polyenes and aromatics. The chemical structure of photosensing pigment/protein complexes has been resolved for many of the photobiological processes that have a characteristic sensitivity in the visible and infrared part of the spectrum of (solar) radiation. However, knowledge about the structure of photoreceptors responsible for several physiologically well-characterized photoresponses to UV- and blue light is still lacking. For a limited number of phototransduction processes, the details of light-induced signal transfer are beginning to be understood in atomic detail. This applies particularly to two photosensors involved in phototactic responses in bacteria: sensory rhodopsin I (SR-I) from Halobacterium salinarium and photoactive yellow protein (PYP) from Ectothiorhodospira halophila. The SR-1 system is of special interest because the transducer accepting the signal from SR-1 was recently identified as Htr-1, a homologue of the methyl-accepting chemotaxis proteins which have been characterized in Escherichia coli. PYP, on the other hand, may be the first photosensor to actually reveal all relevant details of the kinetics, thermodynamics, and molecular motion of light-induced signal generation, through an understanding of how the photo-isomerization of the chromophore forces the sensor protein into the signalling state. Here we compare these photosensors and discuss common themes in the initiation of photosensory signal transduction in microorganisms in terms of the molecular properties of photosensors and their signalling state.

Bacteria↗
PubMed2001

Clinical significance of potential pathogenic microorganisms of sputum in patients with pulmonary tuberculosis.

BACKGROUND: Bacterial culture of sputum is frequently positive in patients with pulmonary tuberculosis (TB). However, it remains to be clarified whether detection of potential pathogenic microorganisms (PPM) in sputum represents bacterial infection or only colonization of the respiratory tract. In the present study, we investigated the clinical significance of PPM in patients with pulmonary TB. METHODOLOGY: Sputum culture for PPM was studied in 174 pulmonary TB patients (117 males and 57 females, mean age of 71 years) on the day of admission. Ninety-seven patients (63 males and 34 females, mean age of 67.1 years) also underwent transtracheal aspirates (TTA). Quantitative culture of sputum and TTA was performed to distinguish infection from colonization. The results were evaluated as follows: more than 106 or 105 c.f.u./mL indicated infection in the case of sputum or TTA samples, respectively. We also compared various clinical parameters between patients with PPM and patients with non-potential pathogenic microorganisms (non-PPM). RESULTS: Potential pathogenic microorganisms were positive in 44% and 24% of sputum and TTA specimens, respectively. However, quantitative culture for PPM showed positive findings in only 11% and 3% of sputum and TTA specimens, respectively. A comparative study of the clinical features revealed that PPM patients had a lower bodyweight and lower serum albumin levels than non-PPM patients. A fatal outcome was also more common in PPM patients than in non-PPM patients. Logistic regression analysis further confirmed that PPM clearly contributed to a fatal outcome in addition to the previously established parameters including age, performance status, haemoglobin, albumin and radiographic disease extent. CONCLUSION: Although sputum PPM represent only colonization of the upper respiratory tract in TB patients, they are associated with a poor prognosis.

Aged↗
PubMed2002

Prevalence and microdiversity of Alteromonas macleodii-like microorganisms in different oceanic regions.

The presence, prevalence and variability of microorganisms related to the species Alteromonas macleodii, a well known culturable gamma-Proteobacterium, has been studied in different seawater samples from diverse geographical locations, in both the Northern and Southern hemispheres, and tested with two molecular techniques (rRNA hybridization and gene cloning and sequencing). Results show that A. macleodii-like microorganisms are present in high proportions in North Atlantic and, especially, Mediterranean waters, being higher at deep samples and particle-associated fractions, in agreement with previous findings. In contrast, Southern samples (all from very cold areas near Antarctica) presented no significant hybridization signals. The analysis of the ribosomal ITS (16S-23S internal transcribed spacers) revealed that A. macleodii-like microorganisms from Mediterranean, North Atlantic, Caribbean and Red Sea waters differed in both size and sequence, mostly depending on their geographical origin, with Mediterranean and North Atlantic clones clustering into two main groups whereas Caribbean and Red Sea clones appeared separated.

Alteromonas↗
PubMed2004

Microorganisms and volatile organic compounds in airborne dust from damp residences.

Airborne dust samples from damp (n = 9) and control (n = 9) residences were analyzed for microorganisms (molds and bacteria), bacterial markers (3-hydroxy fatty acids and muramic acid), and adsorbed volatile organic compounds (VOCs). The number of mold species was greater in the damp residences than in the controls (23 vs.18) and nine mold species were found only in damp residences. The levels of 3-hydroxy fatty acids and muramic acid correlated better in damp residences than in controls, indicating that damp conditions affect the bacterial flora of airborne dust. Identifications made by culture and microscopy of the major molds found, i.e. Aspergillus, Cladosporium, and Penicillum, coincided with the identification of VOCs known to be produced by these species. A number of additional VOCs irritating to the skin, eyes, or respiratory tract were also found. The results from this pilot study illustrate the diversity of microorganisms and VOCs present in the indoor environment and suggest that analysis of airborne dust may help to assess human exposure to microorganisms and chemical compounds.

Air Pollution, Indoor↗
PubMed1997

Utilization of algal polysaccharides by human colonic bacteria, in axenic culture or in association with hydrogenotrophic microorganisms.

The ability of different hydrolytic bacteria from the human colon to grow on various algal polymers (carrageenans, Palmaria palmata xylan, ulvan, desulphated ulvan and laminaran) was investigated and the interactions between Bacteroides thetaiotaomicron and H2-utilizing microorganisms (one methanogenic archaea and an acetogenic bacterium) were studied during laminaran degradation. None of the algal polysaccharides supported the growth of any of the hydrolytic species tested, except for laminaran, which allowed substantial growth of B thetaiotaomicron. This suggested that bacterial consortia were involved in algal polymer breakdown rather than one specific bacterial species. The presence of H2-utilizing microorganisms did not increase the extent of laminaran degradation by B thetaiotaomicron. Whereas the decrease in formate and H2 concentrations attested to their utilization by both hydrogenotrophic microorganisms, the large increase in acetate production observed in the coculture with acetogenic bacteria was mainly due to acetogenic fermentation of sugars released during laminaran hydrolysis.

Bacteroides↗
PubMed2005

Utilising the synergy between plants and rhizosphere microorganisms to enhance breakdown of organic pollutants in the environment.

BACKGROUND: Phytoremediation is a promising technology for the cleanup of polluted environments. The technology has so far been used mainly to remove toxic heavy metals from contaminated soil, but there is a growing interest in broadening its applications to remove/degrade organic pollutants in the environment. Both plants and soil microorganisms have certain limitations with respect to their individual abilities to remove/breakdown organic compounds. A synergistic action by both rhizosphere microorganisms that leads to increased availability of hydrophobic compounds, and plants that leads to their removal and/or degradation, may overcome many of the limitations, and thus provide a useful basis for enhancing remediation of contaminated environments. MAIN FEATURES: The review of literature presented in this article provides an insight to the nature of plant-microbial interactions in the rhizosphere, with a focus on those processes that are relevant to the breakdown and/or removal of organic pollutants. Due consideration has been given to identify opportunities for utilising the plant-microbial synergy in the rhizosphere to enhance remediation of contaminated environments, RESULTS AND DISCUSSION: The literature review has highlighted the existence of a synergistic interaction between plants and microbial communities in the rhizosphere. This interaction benefits both microorganisms through provision of nutrients by root exudates, and plants through enhanced nutrient uptake and reduced toxicity of soil contaminants. The ability of the plant-microbial interaction to tackle some of the most recalcitrant organic chemicals is of particular interest with regard to enhancing and extending the scope of remediation technologies. CONCLUSIONS: Plant-microbial interactions in the rhizosphere offer very useful means for remediating environments contaminated with recalcitrant organic compounds. OUTLOOK: A better knowledge of plant-microbial interactions will provide a basis for improving the efficacy of biological remediations. Further research is, however, needed to investigate different feedback mechanisms that select and regulate microbial activity in the rhizosphere.

Biodegradation, Environmental↗
PubMed1998

Antibacterial effects of a bioactive glass paste on oral microorganisms.

Bioactive glasses contain oxides of calcium, sodium, phosphorus, and silicon in a proportion that provides the material with surface activity and concomitantly with the property of forming a strong bond with bone. Bioactive glasses have been tested as bone substitutes in different clinical situations. In an aqueous environment, Ca2+, Na+, PO4(3-) , and Si4+ are released from the glass, resulting in a rise in pH and in osmotic pressure in its vicinity. Since these are factors that potentially influence the viability of oral microorganisms at the dentogingival margin, we studied the effects of bioactive glass S53P4 on the oral microorganisms Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Actinomyces naeslundii, Streptococcus mutans, and Streptococcus sanguis. This was done by incubating each microbe in a suspension, in the presence of bioactive glass S53P4 in powder form. A. naeslundii was found to lose its viability within 10 min under the experimental conditions. A. actinomycetemcomitans, P. gingivalis, and S. mutans lost their viability within 60 min. Also for S. sanguis a significant loss of viability was seen within 60 min, but it was the only microbe that had any viable cells left after 60 min. Thus, in aqueous solutions the powdered bioactive glass S53P4 appears to have a broad antimicrobial effect on microorganisms of both supra- and subgingival plaque. Consequently, it could be useful as an ingredient in tooth-care products that may have beneficial effects on oral health both from a cariologic and a periodontal point of view.

Actinomyces↗
PubMed1998

Different pharmacokinetics of (4R)-hexahydro-7,7-dimethyl-6-oxo-1,2,5-(3-14C)dithiazocine-4-carboxyli c acid between the fasting and non-fasting rat: role of intestinal microorganisms.

1. We report differences in the pharmacokinetics of (4R)-hexahydro-7,7-dimethyl-6-oxo-1,2,5-(3-14C)dithiazocine-4-carb oxylic acid (14C-SA3443) between the normal fasting and non-fasting rat, especially in the blood concentration-time curves and respiratory excretion. Exhalation of 14CO2 was an important route of elimination and accounted for 21.2% of the dose in the non-fasting rat but only 3.7% in fasting animals. 2. In the intestinal microorganism-compromised rat, we found little differences in the pharmacokinetics of 14C-SA3443 between fasting and non-fasting states. No respiratory excretion was observed in the intestinal microorganism-compromised animal. 3. In the reaction mixture of 14C-SA3443 with the cecal contents of rat, 14C-acetic acid and 14C-butyric acid were detected and 14CO2 barely detected. 4. The amounts of 14C-acetic acid and 14C-butyric acid in the reaction mixture of 14C-SA3443 with non-fasting rat cecal contents were more than those with fasting rat cecal contents. 5. We concluded that the reason for the different pharmacokinetics of 14C-SA3443 between the fasting and non-fasting rat was the differences in participation of the metabolism of 14C-SA3443 by intestinal microorganisms.

Adjuvants, Immunologic↗
PubMedDate not supplied

Unidentified bacterial microorganisms entrapped within blood capillary spaces of tissue from different epidemiological types of Kaposi's sarcoma.

Tissue specimens of different epidemiological types of Kaposis sarcoma (KS) from various geographical regions were investigated by transmission electron microscopy. Freshly fixed KS biopsies originated from 9 German patients: 3 classic KS cases, 5 AIDS-associated KS cases, and 1 atypical classic KS case. Additionally, KS autopsy material from the brain of a German AIDS patient was examined. Further biopsies came from 29 Ugandan patients: 16 endemic KS cases and 13 AIDS-associated KS cases. While investigating the ultrastructure, we discovered relatively small-sized bacterial microorganisms within blood capillary spaces of tumor tissue from 5 KS cases of different epidemiological type. The microorganisms often occurred in clusters. They were of coccoid-bacillary form and limited by a wrinkled multilayered cell wall. Many of them were encapsulated. They were not observed outside of the capillary lumen. The bacterial structures were often seen attached to capillary endothelial cells, which sometimes showed blistering into the capillary lumen. The observed bacterial microorganisms obviously represented agents of a bloodstream infection and must have been entrapped and accumulated within capillary spaces of KS tissue. The bacteria, which had an almost identical morphology in all 5 KS cases, could not be identified. If they are of pathogenic significance, it remains unknown.

Acquired Immunodeficiency Syndrome↗
PubMedDate not supplied

Identifying the determinants of viable microorganisms in the air and bulk metalworking fluids.

Exposure assessment was conducted for an epidemiologic study of the respiratory effects of exposure to metalworking fluids (MWF). As part of the study, airborne microorganisms were collected with a two-stage microbial impactor, and a sample of the bulk soluble MWF was collected from each machine sump, as well as information about the work environment. These data were then used to develop multivariate statistical models of the determinants bulk MWF and airborne microbial levels. Microbial concentrations in the bulk MWF ranged from 5 x 10(4) to 5 x 10(10) colony-forming units (CFU)/mL, with a geometric mean of 3.4 x 10(7) CFU/mL. The geometric mean airborne microbial level was 182 CFU/m3 (for particles size <8 microm) with a range of 1 to 8,308 CFU/m3. In modeling the determinants of bulk microorganisms, fluid-related factors were the most important characteristics associated with microbial levels, followed by process-related and environmental factors. The final full multivariate model predicted a significant reduction in bulk microbial levels by increasing pH of the fluid and reducing the amount of tramp oil leaking into the fluid. For the airborne microbial models, process-related factors were the major characteristics associated with microbial levels, followed by factors related to worker activities and environmental factors. The final full multivariate model predicted a significant control of airborne microorganisms by increasing worker distance from the machine, reducing the number of machines within 10 feet of the worker, decreasing the bulk microbial levels, and adding machine enclosures. These models can be used to prioritize nonbiocidal interventions to control microbial contamination of the bulk MWF and the air.

Air Microbiology↗
PubMed2000

Fermentation of carbohydrates and yield of microbial protein in mixed cultures of rabbit caecal microorganisms.

Fermentation pattern and yields of microbial protein were investigated in cultures of the rabbit caecal contents supplied with glucose, xylose, starch, pectin and xylan. Rabbits at the age of 4 weeks (before weaning) and 3 months were slaughtered, their caecal contents added at 1.1% to growth media and incubated anaerobically at 39 degrees C for 18 h. Caecal microorganisms of 4-week-old rabbits produced no methane and caproate, less butyrate, but more propionate than microorganisms of 3-month-old rabbits. In both groups of rabbits, fermentation of xylose produced significantly more propionate and less butyrate than fermentation of glucose. More propionate and less acetate was formed from starch than from pectin. In caecal cultures from 4-week-old rabbits with pectin, the molar percentages of acetate was significantly higher and percentages of other short-chain fatty acids (SCFA) lower than in cultures with starch or xylan. In cultures from 3-month-old rabbits, fermentation of pectin and xylan produced similar SCFA profiles, different from SCFA molar composition in cultures with starch. Average production of microbial protein was 129 mg per 1 g of carbohydrate digested (range 110 to 141 mg/g). Protein yields were the same on glucose and xylose, but nonsignificantly higher on starch than on pectin and xylan. It can be concluded that the characteristics of substrate affected fermentation pattern in mixed cultures of rabbit caecal microorganisms. Substrate effects on protein yields were not statistically significant, due to high variation.

Age Factors↗
PubMed2001

PCB biodegradation in aged contaminated soil: interactions between exogenous Phanerochaete chrysosporium and indigenous microorganisms.

This work investigated whether the interaction between the white-rot fungus Phanerochaete chrysosporium and indigenous microorganisms could enhance polychlorinated biphenyl (PCB) removal from historically contaminated soil in aerobic microcosms. The PCB mixture was composed mainly of 14% tri-, 20% tetra-, 9% penta-, 17% hexa-, 26% hepta-, 11% octa-, and 3% nona-chlorobiphenyl (CB) congeners, determined by GC/MS. The fungus, which was grown on sugarcane bagasse and added via this solid substrate, successfully colonized the contaminated soil. The added fungi and the indigenous soil community biodegraded most PCB congeners, with removing efficiencies ranging from 13% to 100% for the 45-day incubation period. The interaction between the fungus and the microorganisms present in the added bagasse inhibited both heterotrophic activity (measured by CO2 evolution) and PCB degradation, suggesting a possible antagonism. In contrast, analysis of variance (ANOVA) inferred a synergistic effect between fungus and soil microorganisms, which resulted in a heterotrophic activity above 2.5 mg-CO2/g-initial dry matter/day. The statistical analyses also showed that the presence of fungus alone was particularly beneficial for the removal of penta- and hepta-CB.

Biodegradation, Environmental↗
PubMed2001

Removal of pathogenic and indicator microorganisms by a constructed wetland receiving untreated domestic wastewater.

Wetlands containing floating, emergent and submergent aquatic plants, and other water-tolerant species have been found to economically provide a mechanism of enhancing the quality of domestic wastewater. The use of constructed wetlands for the removal of indicator bacteria (total and fecal coliforms), coliphages, protozoan parasites (Giardia and Cryptosporidium) and enteric viruses was investigated. A pilot scale constructed wetland consisting of two cells, one planted with bulrush and the other unplanted bare sand, were used to compare their efficiency in removing pathogens from raw sewage. Overall more than 90 percent of all microorganisms studied were removed by either of the two systems with a 1 to 2 day retention time. Removal of all mentioned microorganisms was greater from the surface flow in the unplanted cell than in the planted cell, except for Giardia and Cryptosporidium, although the differences were not statistically significant. Enteric viruses, coliphages and indicator bacteria were found to penetrate 2 m below the surface, although concentrations were reduced by greater than 99 percent in both cells. Less virus penetration into the sand occurred in the planted wetland versus the unplanted wetland. Water temperature was found to be the most important factor in the removal of enteric bacteria and viruses, while turbidity reduction was related to Giardia removal. These results demonstrate that significant reductions of pathogenic microorganisms can occur in constructed wetlands receiving untreated domestic wastewater with only a 1-2 day retention time.

Animals↗
PubMed2001

Transformation of the insecticide teflubenzuron by microorganisms.

Transformation of teflubenzuron, the active component in the insecticide commercialized as Nomolt, by soil microorganisms was studied. It was shown that microorganisms, belonging to Bacillus, Alcaligenes, Pseudomonas and Acinetobacter genera are capable to perform the hydrolytic cleavage of the phenylurea bridge of teflubenzuron in different positions, especially active was Bacillus brevis 625. The structure of the intermediates formed was established using TLC, HPLC, mass-spectrometry and 19F NMR techniques. It was shown that for a dose range of 53-132 microM and upon 12 days of fermentation about 30% of the teflubenzuron was modified. About 10-15% was transformed into 2,6-difluorobenzamide, 3-5% into 2,6-difluorobenzoic acid, 10-12% into 2,4-difluoro-3,5-dichloro-aniline. The late compound gave rise to formation of a condensed compound, identified as 1,2-bis(2,4-difluoro-3,5-dichlorophenyl)urea with molecular mass of 420. The results obtained indicate degradation of teflubenzuron by soil microorganisms to be a process to be mediated by microbial consortia, and starting with hydrolysis of the phenylurea bridge by several bacterial species. Subsequent further degradation of the aromatic degradation products has to be mediated by other strains known to be capable of degradation of halogenated aromatics.

Acinetobacter calcoaceticus↗
PubMed1960

Degranulation of polymorphonuclear leucocytes following phagocytosis of microorganisms.

A marked reduction in numbers of cytoplasmic granules in rabbit and human polymorphonuclear leucocytes takes place following ingestion of various microorganisms or of a yeast cell wall preparation. The degranulation occurs within 30 minutes of phagocytosis, and is directly related to the quantity of material engulfed. White cells completely degranulated following phagocytosis of large numbers of microorganisms remain viable for at least 1 hour. The granules of polymorphonuclear leucocytes contain the antimicrobial agent, phagocytin, and various digestive enzymes. These substances thus are released into the cytoplasm or into vacuoles following ingestion of foreign material. The granule system and granule lysis mechanism may well play a central role in the primary function of these specialized cells; namely, that of destroying invading microorganisms.

Animals↗