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Results for “Microbial Viability”

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PubMed1996

Assays for microbial contamination and DNA analysis based on electrorotation.

A new assay is described for determining the concentration and viability of waterborne micro-organisms. The method exploits the fact that when a latex bead coated with a specific binding agent is complexed to the target analyte, the analyte complex formed assumes new dielectric properties that can be monitored by its electrorotation response. This generic technology is applicable to a wide range of organism and toxicological diagnostic tests, as well as to other areas of biotechnology. An example is given of its possible application to DNA sequence analysis.

Animals↗

PubMedDate not supplied

[Viability of facultative anaerobic bacteria in commercial transport systems].

The possibilities of the transport systems manufactured by Copan (Italy) and Deltalab (Spain) were studied on 15 microbial strains: representatives of the family Enterobacteriaceae (enterobacterial swabs with Cary-Blair medium) and the genus Streptococcus, as well as the species Neisseria meningitidis, Haemophilus influenzae (universal swabs with charcoal-enriched Amies medium). Microorganisms were shown to retain their viability (in colony-forming units, %) for 48 hours in the systems of both firms. H. influenzae exhibited greater viability in the system manufactured by Copan than in that manufactured by Deltalab (respectively, 62% and 28% in 24 hours, 19% and 6% in 48 hours).

Anaerobiosis↗

PubMed1997

Effects of sanguinarium, chlorhexidine and tetracycline on neutrophil viability and functions in vitro.

The effectiveness of an ideal antimicrobial agent depends on its ability to kill microbes with minimal toxicity to host cells. Depending on the treatment regimen, antimicrobial agents come into contact with host cells for various intervals of time. Sanguinarium (SANG), chlorhexidine (CHX) and tetracycline (TET) are 3 antimicrobial agents frequently used in the management of periodontal infections. However, their effects on host immune cells during different treatment regimens are not known. Due to their ability to serve as the first line of host defense against microbial infections, we have compared the effects of these antimicrobial agents on human neutrophil functions and viability. The results show that SANG is not lytic to neutrophils from peripheral blood or crevicular fluid, at all concentrations tested. However, exposures of neutrophils to very low concentrations of SANG (0.001%) inhibits neutrophil chemotaxis, oxidative metabolism and degranulation within 5 min. Increasing the exposure time results in a similar inhibition of neutrophil functions, albeit at 50-100 fold lower concentrations of SANG. CHX rapidly disrupts the cell membrane of both crevicular and peripheral blood neutrophils at concentrations above 0.005% within 5 min, and inhibition of all neutrophil functions is due to its lytic properties. While TET is least toxic to neutrophils, a dose dependent inhibition of neutrophil functions is dependent on the calcium concentrations of the cellular environment, and is observed only above 0.04% or higher concentrations in the absence of calcium. The data suggest that a critical cumulative concentration of these drugs is essential for their toxicity and inhibition of neutrophil functions. Therefore, both the length of exposure and the dose of the drug both are critical while considering the effectiveness of SANG, CHX or TET in the treatment of infections. Furthermore, due to differences in their mechanisms of action, the consequences of their effects on neutrophils may have significant bearing on tissue pathology as well as on their therapeutic efficacy.

Adult↗

PubMed2026

PGPR inoculation and growth enhancement of crops cultivated in hydroponic systems.

Plant growth-promoting rhizobacteria (PGPR) are ubiquitous rhizosphere microorganisms that promote plant health through various mechanisms. Although the study of PGPR inoculants in soil has been done for ages, their application in hydroponic systems has received relatively limited attention. This review identifies PGPR inoculants that are commonly used in hydroponics, methods of application, and their effects on plant growth and nutrient use efficiency. Literature shows that PGPR inoculants improve plant performance in controlled hydroponic systems through the production of growth-stimulating substances, nitrogen fixation, and improved nutrient acquisition. However, the plant growth responses are highly variable depending on the composition of nutrient solutions, environmental factors, crop and microbe species, and the type of hydroponic system. The review identifies various challenges of PGPR inoculation in hydroponic systems and future research directions to address the current gaps. Generally, the productivity of hydroponic systems can be enhanced through advanced inoculation strategies and the development of suitable carrier materials to improve inoculant survival, viability, and functions. Emphasis should also be placed on designing system-specific microbial consortia and Synthetic communities that are tailored to the unique ecological conditions of hydroponic systems.

Hydroponics↗

PubMed2006

Antimicrobial function of Nd3+-doped anatase titania-coated nickel ferrite composite nanoparticles: a biomaterial system.

The present study describes and makes a relative comparison of the antimicrobial function of undoped and neodymium-doped titania coated-nickel ferrite composite nanoparticles processed by uniquely combining the reverse micelle and chemical hydrolysis approaches. This methodology facilitates the formation of undoped and doped photocatalytic titania shells and a magnetic ferrite core. The ferrite core is needed to help in the removal of particles from the sprayed surface using a small magnetic field. Doping of the titania shell with neodymium significantly enhances the photocatalytic and anti-microbial function of the core-shell composite nanoparticles without influencing the magnetic characteristics of the nickel ferrite core. The increased performance is believed to be related to the inhibition of electron-hole recombination and a decrease in the band gap energy of titania. The retention of magnetic strength ensures controlled movement of the composite nanoparticles by the magnetic field, facilitating their application as removable anti-microbial photocatalyst nanoparticles. The consistent behavior of the composite nanoparticles points to the viability of the synthesis process adopted.

Anti-Infective Agents↗

PubMed1996

Granulocytes harvested following G-CSF-enhanced leukocyte recovery retain their functional capacity during in vitro culture for 72 hours.

The purpose of this study was to compare two different in vitro culture conditions for the preservation of human granulocytes. These cells could be used in patients with severe neutropenia following cytotoxic chemotherapy if the functional capacity was retained, and autologous transfusions of granulocytes would circumvent the risk of alloimmunization. Granulocytes were obtained from the peripheral blood of healthy donors and patients with hematologic malignancies who received cytotoxic chemotherapy supported by recombinant human granulocyte colony-stimulating factor (R-metHuG-CSF, 300 micrograms/day, s.c.). Granulocytes were either cultured for 72 h at 4 degrees C in the presence of 100 ng/ml G-CSF or cryopreserved at -196 degrees C. The viability, surface antigen expression, and function of the granulocytes were assessed. Since effective microbial killing involves the attachment of granulocytes to blood vessel walls, transmigration into tissues, chemotaxis, and phagocytosis, the surface expression of the adhesion molecules LFA-1 (CD11a/CD18) and gp 150,95 (CD11c/CD18) was measured. In addition, the IgG receptors Fc gamma RI (CD64), Fc gamma RII (CD32), and Fc gamma RIII (CD16), as well as the complement receptor CR3 (CD11b/CD18), were assessed. Dynamic superoxide anion release served as a measure of the metabolic pathway of the oxidative burst after f-Met-Leu-Phe (fMLP) and phorbol-12-myristate-13-acetate (PMA) stimulation. Substantial differences in the preservation of granulocyte integrity and function were observed between the two storage conditions. Cryopreservation abolished reactivity to extracellular stimuli and severely affected the cell phenotype. On the other hand, functional activity could be maintained for up to 72 h when in vivo primed granulocytes of patients were incubated at 4 degrees C in the presence of G-CSF. This storage modality may permit the use of granulocyte autotransfusion to reduce the risk of neutropenic fever.

Antigens, Surface↗

PubMed2003

Cutting edge: Mycobacterium tuberculosis blocks Ca2+ signaling and phagosome maturation in human macrophages via specific inhibition of sphingosine kinase.

One-third of the world's population is infected with Mycobacterium tuberculosis (Mtb), and three million people die of tuberculosis each year. Following its ingestion by macrophages (MPs), Mtb inhibits the maturation of its phagosome, preventing progression to a bactericidal phagolysosome. Phagocytosis of Mtb is uncoupled from the elevation in MP cytosolic Ca(2+) that normally accompanies microbial ingestion, resulting in inhibition of phagosome-lysosome fusion and increased intracellular viability. This study demonstrates that the mechanism responsible for this failure of Ca(2+)-dependent phagosome maturation involves mycobacterial inhibition of MP sphingosine kinase. Thus, inhibition of sphingosine kinase directly contributes to survival of Mtb within human MPs and represents a novel molecular mechanism of pathogenesis.

Animals↗

PubMed1983

Growth of bacteria in intravenous fluids under stimulated actual-use conditions.

The growth of microorganisms in nonnutritive intravenous solutions under simulated actual-use conditions was studied. Small quantities of Pseudomonas aeruginosa, Staphylococcus aureus, and Klebsiella pneumoniae (final concentration 200-400 cells/ml) were injected into 500-ml containers (glass bottles and plastic bags) of 5% dextrose injection, 0.9% sodium chloride injection, and 5% dextrose and 0.9% sodium chloride injection. Additives (ampicillin, vitamin K, lidocaine, and vitamin B complex) were included in some i.v. solutions. Administration sets were attached to the i.v. containers, and the solutions were run into collection bottles; samples were withdrawn at 0, 1, 2, 3, 4, 5, 6, and 8 hours after contamination and plated for viable counts. Staph. aureus and K. pneumoniae remained viable in 5% dextrose injection and in 0.9% sodium chloride injection, but the numbers of these bacteria did not increase. The number of Ps. aeruginosa declined in all three solutions. In 5% dextrose and 0.9% sodium chloride injection, the number of K. pneumoniae declined but Staph. aureus maintained viability. The type of container and the drug additives had no effect on microbial growth, except that ampicillin was bactericidal to Staph. aureus. Low-level contamination of these bacteria in nonnutritive i.v. solutions under actual-use conditions does not result in large numbers of organisms within the time frame in which most solutions are administered.

Bacteria↗

PubMed2004

Microbial astronauts: assembling microbial communities for advanced life support systems.

Extension of human habitation into space requires that humans carry with them many of the microorganisms with which they coexist on Earth. The ubiquity of microorganisms in close association with all living things and biogeochemical processes on Earth predicates that they must also play a critical role in maintaining the viability of human life in space. Even though bacterial populations exist as locally adapted ecotypes, the abundance of individuals in microbial species is so large that dispersal is unlikely to be limited by geographical barriers on Earth (i.e., for most environments "everything is everywhere" given enough time). This will not be true for microbial communities in space where local species richness will be relatively low because of sterilization protocols prior to launch and physical barriers between Earth and spacecraft after launch. Although community diversity will be sufficient to sustain ecosystem function at the onset, richness and evenness may decline over time such that biological systems either lose functional potential (e.g., bioreactors may fail to reduce BOD or nitrogen load) or become susceptible to invasion by human-associated microorganisms (pathogens) over time. Research at the John F. Kennedy Space Center has evaluated fundamental properties of microbial diversity and community assembly in prototype bioregenerative systems for NASA Advanced Life Support. Successional trends related to increased niche specialization, including an apparent increase in the proportion of nonculturable types of organisms, have been consistently observed. In addition, the stability of the microbial communities, as defined by their resistance to invasion by human-associated microorganisms, has been correlated to their diversity. Overall, these results reflect the significant challenges ahead for the assembly of stable, functional communities using gnotobiotic approaches, and the need to better define the basic biological principles that define ecosystem processes in the space environment.

Bacteria↗

PubMed2005

New methods in the evaluation of chemical disinfectants used in health care services.

BACKGROUND: A considerable number of tests are recommended in the literature to evaluate in vitro commercial chemical solutions. The variety of tests reflects their limitations and the need to enhance disinfection process. METHODS: In this study the efficacy of 4 chemical disinfectants selected by their practical use in Health Care Services and by literature recommendation in aerobic and in strict anaerobic bacteria were evaluated by their practical use in Health Care Services and by literature recommendation in aerobic and in strict anaerobic bacteria. Viability was tested in biofilms grown on glass and rubber tip carriers. RESULTS: The results showed microbial growth in chemical solutions at concentrations recommended by the literature or at very close concentrations to them. Viable cells were recovered from biofilms after 30 minutes (Bacteroides fragilis) and 60 minutes (Streptococcus mutans and Salmonella tiphymurium) contact with 2.4% glutaraldehyde and after 60 minutes (S tiphymurium) in 2.0% glutaraldehyde. In 70% ethyl alcohol, S tiphymurium was viable up to 10 minutes, Escherichia coli up to 30 minutes, and S mutans up to 60 minutes. In 1% sodium hypochlorite, S mutans was viable up to 30 minutes and S tiphymurium up to 45 minutes. Detection of cell viability could be related to methodologic differences, including biofilm formation, as demonstrated by scanning electron microscopy. It should be emphasized that B fragilis, the most clinically relevant obligate anaerobe, remained viable in one routinely used solution. CONCLUSION: These findings pointed out the need of periodic surveillance of disinfectants' activity used in Health Care Services and the need of reviewing routines of disinfection protocols.

Bacteria↗

PubMed1990

The effect of antibiotics that inhibit cell-wall, protein, and DNA synthesis on the growth and morphology of Legionella pneumophila.

The response of Legionella pneumophila to antibiotics that inhibit cell-wall, protein and DNA synthesis was examined by electronmicroscopy, MIC estimations and viable counts. Ampicillin, cefotaxime, methicillin, erythromycin, rifampicin and ciprofloxacin, each used separately at 20 times their respective MIC values, showed activity against L. pneumophila in these studies. The inhibitors of cell-wall synthesis--ampicillin, cefotaxime and methicillin--effected the greatest bactericidal activity and induced the most extensive morphological changes, which included the formation of membranous lesions through which cytoplasmic contents were lost. In terms of ultrastructural damage and loss of viability, the inhibitors of protein and DNA synthesis were less effective than the antibiotics that acted on the microbial cell wall. Erythromycin- and rifampicin-treated cells possessed irregular membranes and were partially or fully lysed, whereas ciprofloxacin induced abnormally elongated organisms with intermittently lysed and detached inner membranes. These results illustrated the ability of antibiotics of putative clinical value, with diverse modes of action, to affect the ultrastructural cytology as well as the viability of L. pneumophila in vitro.

4-Quinolones↗

PubMed2005

Molecular mimicry: anti-DNA antibodies bind microbial and nonnucleic acid self-antigens.

Although cells of the innate immune response have a variety of pattern recognition receptors that are triggered by blood classes of markers, a critical feature of the adaptive immune response is antigenic specificity. Yet it is becoming increasingly clear that the specificity of lymphocyte receptors admits of some laxity. Cross-reactivity may, in fact, be necessary for lymphocyte survival as antigen receptor signaling maintains cellular viability in the absence of antigen activation. Studies of molecular mimicry have revealed many instances in which antibodies to microbial antigens bind also to self-antigens; in some cases, this cross-reactivity has pathogenic potential. In this chapter, we describe cross-reactivity between two self-antigens, DNA and NMDA receptors, and how antibodies with specificity for DNA in patients with splenic lupus may cause central nervous system damage by virtue of binding also to neuronal receptors. This example serves as a reminder that cross-reactivity may exist among self-antigens as well as between foreign and self-antigens.

Animals↗

PubMed1999

Effect of ultraviolet light irradiation and nitrosoguanidine on viability of 46 strains of Arthrinium and their antibiotic production.

The ability of strains of the Arthrinium genus to inhibit microbial development has been previously described. In the present work different periods of mutagenic treatment using ultraviolet light, and of nitrosoguanidine treatment, on strains of Arthrinium were investigated. With nitrosoguanidine treatment the survival rate ranged from 2.17 to 8.78%. Mutant strains were only obtained with a higher antibiotic production in comparison with the wild-strain, when the mutagenic agent was UV light.

Anti-Bacterial Agents↗

PubMed1992

The endodontic microflora revisited.

The microbial flora of 35 dental root canals were examined, taking care to maintain the viability of obligate anaerobes which accounted for 45% of total isolations, while streptococcal species accounted for 24% of the total species isolated. Individual root canals yielded a maximum of eight bacterial species. A total of 40 different species was isolated of which the most prevalent were the facultative anaerobe Streptococcus sanguis and the obligate anaerobe, Peptostreptococcus micros (both in 23% of root canals), followed by Eubacterium aerofaciens and the 'Streptococcus milleri group' (both 17%) then Prevotella melaninogenica (formerly Bacteroides melaninogenicus), Enterococcus faecalis and Prevotella oralis (formerly Bacteroides oralis), which were each isolated from 14% of root canals. Highly significant associations were discovered between four pairs of species, viz P. melaninogenica with P. micros, P. melaninogenica with P. oralis, Prevotella corporis with Streptococcus morbillorum and Actinomyces odontolyticus with E. faecalis.

Dental Pulp Cavity↗

PubMed1996

Microbial contamination of antiseptics and disinfectants.

BACKGROUND: There have been a number of reports on microbial contamination of antiseptics and disinfectants. At present, however, the necessity of measures to prevent contamination do not seem to be fully appreciated. We investigated microbial contamination of antiseptics and disinfectants that are used in our hospital. METHODS: Fifty-one samples of benzalkonium chloride and chlorhexidine gluconate that were being used in the hospital were examined. Viability of the contaminants detected in these samples was also tested in the agents. Then we examined measures to prevent contamination of these agents. RESULTS: Microbial contamination was detected at 10(2) to 10(7) CFU/ml in the following samples: 6 of 23 samples of cotton balls soaked in 0.02% benzalkonium chloride kept in a canister for antisepsis and disinfection (26.1%); 7 of 13 samples of 0.02%, benzalkonium chloride or 0.02% chlorhexidine gluconate in an irrigation apparatus kept at 37 degrees C for vaginal douching (53.8%); and 9 of 15 samples of 0.02% benzalkonium chloride or 0.05% chlorhexidine gluconate for storage of suction catheters in a plastic bottle (60%). The major contaminants were Burkholderia cepacia, Pseudomonas aeruginosa, Xanthomonas maltophilia, and Pseudomonas fluorescens. The first two organisms examined grew in the agents. After improvements in the handling of the antiseptics and disinfectants, no microbial contamination was observed. CONCLUSIONS: It is necessary to check microbial contamination of diluted benzalkonium chloride and diluted chlorhexidine gluconate that are in use. Such products are not recommended as antiseptics.

Anti-Infective Agents, Local↗

PubMed2006

Selective removal of DNA from dead cells of mixed bacterial communities by use of ethidium monoazide.

The distinction between viable and dead bacterial cells poses a major challenge in microbial diagnostics. Due to the persistence of DNA in the environment after cells have lost viability, DNA-based quantification methods overestimate the number of viable cells in mixed populations or even lead to false-positive results in the absence of viable cells. On the other hand, RNA-based diagnostic methods, which circumvent this problem, are technically demanding and suffer from some drawbacks. A promising and easy-to-use alternative utilizing the DNA-intercalating dye ethidium monoazide bromide (EMA) was published recently. This chemical is known to penetrate only into "dead" cells with compromised cell membrane integrity. Subsequent photoinduced cross-linking was reported to inhibit PCR amplification of DNA from dead cells. We provide evidence here that in addition to inhibition of amplification, most of the DNA from dead cells is actually lost during the DNA extraction procedure, probably together with cell debris which goes into the pellet fraction. Exposure of bacteria to increasing stress and higher proportions of dead cells in defined populations led to increasing loss of genomic DNA. Experiments were performed using Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium as model pathogens and using real-time PCR for their quantification. Results showed that EMA treatment of mixed populations of these two species provides a valuable tool for selective removal of DNA of nonviable cells by using conventional extraction protocols. Furthermore, we provide evidence that prior to denaturing gradient gel electrophoresis, EMA treatment of a mature mixed-population drinking-water biofilm containing a substantial proportion of dead cells can result in community fingerprints dramatically different from those for an untreated biofilm. The interpretation of such fingerprints can have important implications in the field of microbial ecology.

Affinity Labels↗

PubMed1993

Application of flow cytometry to rapid microbial analysis in food and drinks industries.

In food and drinks industries, the time required for conventional tests can lead to substantial delays in product release to the market. Flow cytometry (FCM) has been used in conjunction with viability markers for rapid counting of yeast, mould and bacterial cells in food products. A single-parameter flow cytometer has proved applicable to the rapid detection of low numbers of microbial contaminants in finished products. The excellent correlation between FCM results and product quality shelf-life expiry date has allowed the establishment of realistic quality control criteria for rapid positive release of product. Used for the monitoring of microbial biomass during manufacturing processes, flow cytometry allowed a direct assessment of bacterial growth. The reproducibility of the results and the proven correlation with standard plate count method obtained in industrial conditions make FCM a good predictive method for product and process quality control.

Bacteria↗

PubMed1999

The Erlanger silver catheter: in vitro results for antimicrobial activity.

The antimicrobial activity of a silver-impregnated polymer catheter (the Erlanger silver catheter) was demonstrated by determining the microbial adhesion to the surface of the catheter and by measuring the rate of proliferation (viability) of microorganisms at this site. On the surface of a catheter impregnated with silver, according to previously described methods, the bacterial adhesion of Staphylococcus epidermidis is reduced by 28-40%. Bacterial proliferation on the surface of the catheter and biofilm production are also substantially reduced by the elution of free silver ions from the catheter matrix. Bacteriostatic and bactericidal activities can be determined. The antimicrobial efficacy of the silver catheter is not reduced by blood components. There is no loss in antimicrobial activity for weeks after preincubation in water or phosphate buffered saline. The antimicrobial activity depends on the extent of the active silver surface.

Animals↗