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PubMed1994

The immunological adjuvant and vaccine carrier properties of liposomes.

Animal immunization studies by numerous laboratories have shown that liposomes promote humoural and cell-mediated immunity to a wide spectrum of bacterial, protozoan and viral antigens as well as tumour cell antigens, venoms and allergens. Adjuvanticity depends on liposomal structural characteristics which determine vesicle fate in vivo and, thus, the mode of antigen interaction with antigen-presenting cells. Adjuvanticity is further promoted by receptor mediated targeting of liposomes to macrophages, or the presence of other adjuvants including cytokines. The immunoadjuvant function of liposomes is supplemented by their ability to act as a carrier for co-entrapped B and T-cell epitopes, thus eliminating the need for a carrier protein. Recently, a technique has been developed for the entrapment of live or attenuated microbial vaccines into giant liposomes under conditions which retain their viability. Liposomes containing microbial vaccines (together with other soluble antigens or cytokines) could be used as carriers of vaccines in cases where there is a need to prevent interaction of vaccines with maternal antibodies or preformed antibodies to vaccine impurities.

Adjuvants, Immunologic↗

PubMed1998

Sonic and mechanical toothbrushes. An in vitro study showing altered microbial surface structures but lack of effect on viability.

The purpose of the present study was to compare the in vitro effects of a mechanical and a sonic toothbrush on the viability of Actinomyces viscosus, the rationale being that induction of irreparable microbial damage resulting from aggressive mechanical action or sonic energy, may inhibit or disrupt the process of successional colonization. Cultures of A. viscosus were grown to a standardized optical density and subdivided into 3 treatment groups of 20 specimens each. Treatment groups consisted of an untreated control and exposure to a mechanical or sonic toothbrush for 15, 30, 45, and 60 s. Subsequent to the prescribed treatment, samples were taken from each specimen dish, subcultured, and the number of CFUs determined. Additional samples were obtained for negative staining and examination by electron microscopy. The mean number of CFUs for each treatment group at each treatment interval were statistically analyzed by ANOVA and multiple pairwise comparisons. Results showed a significant main effect for toothbrushes (p<0.0001) and exposure time (p<0.01), but only marginal significance for the interaction of toothbrush with exposure time (p<0.055). Post-hoc tests showed a significantly greater number of CFUs for the sonic toothbrush compared to both the untreated control and mechanical toothbrush groups. Electron microscopic examination revealed a decrease in aggregation tendency and loss of fimbriae in the sonic toothbrush group. Based on the lack of morphologic evidence that would indicate cell damage and the increase in CFUs over that of the control group, it appeared that neither the mechanical or sonic toothbrushes affected cell viability.

Actinomyces viscosus↗

PubMed1984

In vitro plaque formation on dental amalgam.

A previously described method for the study of in vitro microbial plaque formation on dental materials has been utilized to assess microbial adhesion and growth on five commercially available dental amalgams. Variations in both the amount and viability of the microbial plaques was found between the various types of amalgams.

Adenosine Triphosphate↗

PubMed1988

Viability of microorganisms in fluorouracil and cisplatin small-volume injections.

The ability of various microorganisms to survive in cisplatin injection or fluorouracil injection in small-volume containers was assessed. Inoculum suspensions of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Pseudomonas cepacia, Candida albicans, or Aspergillus niger were injected into polyvinyl chloride minibags containing 20 mL of either fluorouracil injection 50 mg/mL or cisplatin injection 1 mg/mL in 0.9% sodium chloride injection. All minibags were stored at 25 degrees C for 28 days. Three minibags containing only the sodium chloride solution and 18 minibags containing the sodium chloride solution inoculated with microorganisms served as negative and positive controls, respectively. Samples (1 mL) were obtained within 30 minutes of inoculation and again on days 1, 2, 4, 7, 14, 21, and 28. All microbial species lost viability in both the cisplatin and the fluorouracil admixtures over the study period. In most instances, this loss was more pronounced than that in the positive control solution. Whether the pH of the solutions contributed to this loss of viability was not determined. Under the conditions of this study, admixtures containing cisplatin and fluorouracil did not support rapid growth of microorganisms when stored at room temperature for up to 28 days.

Bacteria↗

PubMed1978

Viability of Leptospira interrogans serotype grippotyphosa in swine urine and blood.

Normal swine urine, devoid of its microbial flora, diminished the viability and virulence of Leptospira interrogans serotype grippotyphosa. Bovine serum albumin diluent effectively offset the urine's deleterious effects. Membrane filtration (0.45 micrometer) rendered urine free of bacteria, but permitted passage of limited numbers of leptospires. Most of the commonly used anticoagulants did not alter viability of grippotyphosa in whole blood. However, EDTA significantly reduced the number of viable cells.

Animals↗

PubMed1990

The biochemical mechanisms of the plant activation of promutagenic aromatic amines.

Using specific monooxygenase and oxidase inhibitors in a plant cell/microbe coincubation assay, the biochemical mechanisms of the plant activation of two aromatic amines were compared. The biological endpoints included mutation induction, inhibition of mutagenicity, viability of the plant cells (activating system), and viability of the microbial cells (genetic indicator organism). The activation of m-phenylenediamine by TX1 cells was mediated by enzyme systems that were inhibited by diethyldithiocarbamate, potassium cyanide, methimazole, (+)-catechin or acetaminophen. The inhibition by metyrapone was attended by toxicity in the plant cells. These data implicate a TX1 cell peroxidase and a FAD-dependent monooxygenase in the plant activation of m-phenylenediamine. The TX1 cell activation of 2-aminofluorene was inhibited by diethyldithiocarbamate, 7,8-benzoflavone, acetaminophen or (+)-catechin. An additional pathway of the plant cells in the activation of 2-aminofluorene may involve a cytochrome P-448-type N-hydroxylase.

Acetaminophen↗

PubMed2004

Viability of endolithic micro-organisms in rocks from the McMurdo Dry Valleys of Antarctica established by confocal and fluorescence microscopy.

The rocks of the McMurdo Dry Valleys desert in Antarctica harbour endolithic communities of micro-organisms such as lichens, fungi, cyanobacteria and bacteria. Establishing the physiological status and viability of these microbial colonies in their natural microhabitat has far-reaching implications for understanding the microbial ecology of the harsh environment of this polar desert. Here we describe the use of confocal microscopy and a specific fluorescent probe (FUN-1) to evaluate the metabolic activity of fungal cells. Application of confocal microscopy also served to identify living and dead bacteria or cyanobacteria using the fluorescent assay reagents Live/Dead SYTO 9 and propidium iodide or SYTOX Green, respectively. In addition, through the use of epifluorescence microscopy, live/dead bacteria and cyanobacteria could be detected by estimating fluorescence from their cell components provoked by simultaneously staining with nucleic acids stains such as DAPI and SYTOX Green.

Antarctic Regions↗

PubMed1995

Special needs for disinfectants in food-handling establishments.

Freedom from microbial and foreign body contamination is essential in food production, and the principal means for controlling the surface route of contamination is sanitation (cleaning and disinfection). After the cleaning phase, disinfection plays a crucial role in further reducing microbial numbers and viability. However, disinfectants for use in the food industry may contaminate the product. Therefore, as well as being effective and suitable for factory use, such disinfectants must also be non-toxic and non-tainting. The author describes the usage criteria and operator safety requirements of disinfectants, together with methods to determine taint potential, toxicity and efficacy. Using these parameters, a limited number of disinfectants are judged suitable for general food industry use and these are compared.

Animals↗

PubMed2005

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↗

PubMed2006

Different modes in antibiotic action of tritrpticin analogs, cathelicidin-derived Trp-rich and Pro/Arg-rich peptides.

The cathelicidin-derived antimicrobial tritrpticin could be classified as either Trp-rich or Pro/Arg-rich peptide. We recently found that the sequence modification of tritrpticin focused on Trp and Pro residues led to considerable change in structure and antimicrobial potency and selectivity, but their mechanisms of microbial killing action were still unclear. Here, to better understand the bactericidal mechanisms of tritrpticin and its two analogs, TPA and TWF, we studied their effect on the viability of Gram-positive S. aureus and Gram-negative E. coli in relation to their membrane depolarization. Although TWF more effectively inhibited growth of S. aureus and E. coli than TPA, only a 30 min exposure to TPA was sufficient to kill both bacteria and TWF required a lag period of about 3-6 h for bactericidal activity. Their different bactericidal kinetics was associated with membrane permeabilization, i.e., TWF showed negligible ability to depolarize the cytoplasmic membrane potential of target cell membrane, whereas we observed significant membrane depolarization for TPA. In addition, while TPA caused rapid and large dye leakage from negatively charged model vesicles, TWF showed very little membrane-disrupting activity. Interestingly, we have looked for a synergism among the three peptides against E. coli, supporting that they are working with different modes of action. Collectively, our results suggest that TPA disrupts the ion gradients across the membrane, causing depolarization and a loss of microbial viability. By contrast, TWF more likely translocates across the cytoplasmic membrane without depolarization and then acts against one or more intracellular targets. Tritrpticin exhibits intermediate properties and appears to act via membrane depolarization coupled to secondary intracellular targeting.

Amino Acid Sequence↗

PubMed1997

Evidence that Formation of Protoanemonin from Metabolites of 4-Chlorobiphenyl Degradation Negatively Affects the Survival of 4-Chlorobiphenyl-Cometabolizing Microorganisms.

A rapid decline in cell viability of different PCB-metabolizing organisms was observed in soil microcosms amended with 4-chlorobiphenyl. The toxic effect could not be attributed to 4-chlorobiphenyl but was due to a compound formed from the transformation of 4-chlorobiphenyl by the natural microflora. Potential metabolites of 4-chlorobiphenyl, 4-chlorobenzoate and 4-chlorocatechol, caused similar toxic effects. We tested the hypothesis that the toxic effects are due to the formation of protoanemonin, a plant-derived antibiotic, which is toxic to microorganisms and which has been shown to be formed from 4-chlorocatechol by enzymes of the 3-oxoadipate pathway. Consistent with our hypothesis, addition to soil microcosms of strains able to reroute intermediary 4-chlorocatechol from the 3-oxoadipate pathway and into the meta-cleavage pathway or able to mineralize 4-chlorocatechol by a modified ortho-cleavage pathway resulted in reversal of this toxic effect. Surprisingly, while direct addition of protoanemonin influenced both the viability of fungi and the microbial activity of the soil microcosm, there was little effect on bacterial viability due to its rapid degradation. This rapid degradation accounts for our inability to detect this compound in soils amended with 4-chlorocatechol. However, significant accumulation of protoanemonin was observed by a mixed bacterial community enriched with benzoate or a mixture of benzoate and 4-methylbenzoate, providing the metabolic potential of the soil to form protoanemonin. The effects of soil heterogeneity and microcosm interactions are discussed in relation to the different effects of protoanemonin when applied as a shock load and when it is produced in small amounts from precursors over long periods.

Journal Article↗

PubMed1998

A pilot study on bacterial viability in acne. Assessment using dual flow cytometry on microbials present in follicular casts and comedones.

BACKGROUND: Antibiotic therapy is one of the main methods of acne treatment, however, bacterial resistance is on the rise and can affect the treatment outcome. Quantitative bacteriologic cultures are the gold standard methodology for the assessment of such a problem; however, certain important biological aspects remain uncovered. OBJECTIVE: The purpose of this study was to compare the antibacterial activity of minocycline and lymecycline in sebaceous follicle infundibula and comedones of acne patients. METHOD: We used a recently introduced flow cytometric method, allowing a distinction to be made between viable, injured (presumably resistant), and dead microorganisms. RESULTS: Minocycline (100 mg) proved to be superior to lymecycline (600 mg) in abating the microflora harboring in the sebaceous follicles of acne patients. CONCLUSIONS: The dissimilar bioavailability and antimicrobial efficacy between the two bacteriostatic agents may impart different clinical efficacy.

Acne Vulgaris↗

PubMed2004

A comparison of the effects of ocular preservatives on mammalian and microbial ATP and glutathione levels.

The aim of this study was to investigate the mechanism of action of the preservative sodium chlorite (NaClO2), and the relationship with intracellular glutathione depletion. A detailed comparison of the dose responses of two cultured ocular epithelial cell types and four species of microorganism was carried out, and comparisons were also made with the quaternary ammonium compound benzalkonium chloride (BAK), and the oxidant hydrogen peroxide (H2O2). The viability of mammalian and microbial cells was assessed in the same way, by the measurement of intracellular ATP using a bioluminescence method. Intracellular total glutathione was measured by reaction with 5,5'-dithiobis-2-nitrobenzoic acid in a glutathione reductase-dependent recycling assay. BAK and H2O2 caused complete toxicity to conjunctival and corneal epithelial cells at approximately 25 ppm, in contrast to NaClO2, where > 100 ppm was required. The fungi Candida albicans and Alternaria alternata had a higher resistance to NaClO2 than the bacteria Staphyloccus aureus and Pseudomonas aeruginosa, but the bacteria were extremely resistant to H2O2. NaClO2 caused substantial depletion of intracellular glutathione in all cell types, at concentrations ranging from < 10 ppm in Pseudomonas, 25-100 ppm in epithelial cells, to > 500 ppm in fungal cells. The mechanisms of cytotoxicity of NaClO2, H2O2 and BAK all appeared to differ. NaClO2 was found to have the best balance of high antibacterial toxicity with low ocular toxicity. The lower toxicity of NaClO2 to the ocular cells, compared with BAK and H2O2, is in agreement with fewer reported adverse effects of application in the eye.

Adenosine Triphosphate↗

PubMedDate not supplied

Assessment of viability of yeast by flow cytometry using Light Green as vital dye exclusion.

A comparative study of two different staining methods for estimation of yeast's viability is assessed by microbial flow cytometry. Propidium iodide (PI) a well known nucleic-acid binding vital dye exclusion is compared with light green (LG), a cytoplasmic function related vital dye widely used in histological, histochemical and immunocytochemical staining. The yeast model used was heat-killed Candida guilliermondii cells. The stained heat-killed bacterial population showed on the dot-plot flow cytometer analysis, a clear separation towards the green-orange (350-600 nm) for LG, and orange-red (550-725 nm) for PI on the fluorescence spectrum-zone channels. Both dyes showed by non-parametric Kolmcgorov-Smirnov, 99% of probability of difference between heat-killed and live microorganism's histogram-population, with a D = ranking from 0.50-0.64 for LG to 0.91-0.98 for PI. Higher cost and carcinogenic risk when handling PI, make LG a better choice for flow cytometry yeast viability tests.

Candida↗

PubMed2006

Microbial survival rates of Escherichia coli and Deinococcus radiodurans under low temperature, low pressure, and UV-Irradiation conditions, and their relevance to possible Martian life.

Viability rates were determined for microbial populations of Escherichia coli and Deinococcus radiodurans under the environmental stresses of low temperature (-35 degrees C), low-pressure conditions (83.3 kPa), and ultraviolet (UV) irradiation (37 W/m(2)). During the stress tests the organisms were suspended in saltwater soil and freshwater soil media, at variable burial depths, and in seawater. Microbial populations of both organisms were most susceptible to dehydration stress associated with low-pressure conditions, and to UV irradiation. However, suspension in a liquid water medium and burial at larger depths (5 cm) improved survival rates markedly. Our results indicate that planetary surfaces that possess little to no atmosphere and have low water availability do not constitute a favorable environment for terrestrial microorganisms.

Atmospheric Pressure↗

PubMed1980

A new poultry semen extender. 4. Effect of antibacterials in control of bacterial contamination in chicken semen.

Forty antibacterials were qualitatively and quantitatively tested for controlling aerobic bacterial contamination without affecting viability of semen. Semen samples were collected aseptically, diluted 1:4 with the Beltsville poultry semen extender containing one of 40 antibiotics and held for 0, 24, 48, and 72 hr at 5 C. Semen samples were monitored at each storage interval for bacterial counts, sperm motility, sperm counts, and fertilizing capacity. Gentamicin (2.5 microgram/ml), kanamycin (31.2 microgram/ml), neomycin (62.5 microgram/ml), and tobramycin (2.5 microgram/ml) were the only antibacterials tested which controlled microbial growth without affecting sperm viability for up to 24 hr storage at 5 C. Tobramycin maintained fertility equal to that of the non-antibiotic control up to 24 hr storage. Neomycin maintained higher levels of fertility in semen stored for 48 and 72 hr than in semen of controls to all other treatments for the same storage periods and to undiluted, unstored semen. The conclusion from these studies is that the control of aerobic microbial growth in chicken semen has little influence on the maintenance of sperm viability during low temperature storage.

Animals↗

PubMed2003

Reagentless detection of microorganisms by intrinsic fluorescence.

Quick and accurate detection of microbial contamination is accomplished by a unique combination of leading-edge technologies described in this and the accompanying paper. In this contribution, a hand-held prototype instrument is described which is capable of statistically sampling the environment for microbial contamination and determining cell viability. The technology is sensitive enough to detect very low levels ( approximately 20 cells/cm(2) or cm(3)) of microbes in seconds.

Bacteria↗

PubMed2003

The effect of a toothpaste containing 2% zinc citrate and 0.3% Triclosan on bacterial viability and plaque growth in vivo compared to a toothpaste containing 0.3% Triclosan and 2% copolymer.

OBJECTIVE: To compare the antimicrobial efficacy and effect on plaque growth of a new silica-based fluoride toothpaste containing 2% zinc citrate/ 0.3% Triclosan with a silica-based fluoride toothpaste containing 0.3% Triclosan/2% copolymer. METHODS: In Study 1, plaque was collected after one week's use of each toothpaste and assessed for bacterial viability, live/ dead ratio and microbial membrane integrity. In study 2, plaque was measured immediately and 18 hours after a single brushing with the specified toothpastes. RESULTS: The 2% zinc citrate/0.3% Triclosan formulation significantly reduced the total number of viable aerobic and anaerobic bacteria (p = 0.0223 and p = 0.0443 respectively) compared to the 0.3% Triclosan/2% copolymer formulation. Both toothpastes increased the bacterial membrane permeability significantly. However, the proportion of live bacteria for the 2% zinc citrate/0.3% Triclosan product was significantly reduced (p < 0.05). Study 2 showed significantly less plaque growth 18 hours after using the 2% zinc citrate/0.3% Triclosan toothpaste compared to the 0.3% Triclosan/2% copolymer toothpaste (p < 0.01). CONCLUSION: Regular use of a fluoride toothpaste containing 2% zinc citrate and 0.3% Triclosan, significantly reduced the viability of plaque bacteria compared to a fluoride toothpaste containing 0.3% Triclosan/ 2% copolymer 12 hours after brushing. In addition, a clinical plaque growth study confirmed that this anti-microbial efficacy leads to a significant reduction in plaque growth.

Adult↗