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PubMed2001

Influence of skin maceration and oxygen on anaerobic fermentation of grape musts with high sugar content.

Saccharomyces cerevisiae race cerevisiae was used to ferment grape musts in strictly anaerobic conditions, subjected to a prefermentative treatment of skin maceration and following a short aeration after 48 h of fermentation. Skin maceration caused an increase in the cellular phospholipid content which affected neither viability nor the fermentative capacity of the yeasts. The short aeration had no significant effect on the unsaturation index of the cellular fatty acids, although it did increase the ergosterol/phospholipid ratio. This was reflected by an increase in the growth rate, viability and fermentative capacity of the yeasts. Maceration did not increase the effect of aeration.

Anaerobiosis↗
PubMedDate not supplied

Evaluation of cytotoxicity and antimicrobial activity of Acticoat Burn Dressing for management of microbial contamination in cultured skin substitutes grafted to athymic mice.

Cultured skin substitutes (CSS) have become a useful adjunctive treatment for closure of burn wounds, but CSS are avascular and remain susceptible to microbial destruction longer than split-thickness skin grafts. Irrigation of CSS grafted to burn wounds with a topical antimicrobial solution (TAS) has been shown to promote engraftment of CSS, but TAS usage has potential limitations. Acticoat Burn Dressing (Acticoat; Westaim Biomedical, Exeter, NH) is a silver-coated barrier dressing reported to exhibit antimicrobial activity and to reduce infection in partial-thickness and full-thickness wounds. This study evaluated the cytotoxicity of Acticoat with CSS and the efficacy of Acticoat for the management of microbial contamination in CSS grafted to full-thickness wounds in athymic mice. The cytotoxicity of Acticoat was assessed in preliminary studies after 1 week of exposure to CSS during in vitro maturation or healing on wounds in athymic mice. Histologies were analyzed and cellular viability in the CSS was determined by MTT conversion on days 0, 1, and 7 of Acticoat exposure. At 1, 2, 3, and 4 weeks after grafting, wounds were traced, and areas of healing CSS were calculated by image analysis. At 4 weeks, wound biopsies were evaluated and scored for engraftment of human cells. In a subsequent study, wounds were inoculated with strain SBI-N of Pseudomonas aeruginosa at 1 x 10(5) cfu/wound before the application of CSS or inoculated onto the surface of Acticoat. At 4 weeks, swab cultures were collected from the surface of CSS and scored for the presence of SBI-N. Statistical significance was accepted at the 95% confidence level (P <.05). The data show that exposure in vitro of CSS to Acticoat was cytotoxic within 1 day, but 1 week of exposure in vivo did not injure CSS or inhibit wound healing. Contaminated wounds treated with Acticoat healed similarly to control treatments, with comparable rates of engraftment, and detection of SBI-N on the surface of only one graft. No SBI-N was detected on CSS after inoculation onto the surface of Acticoat. These results suggest that Acticoat may be suitable as a protective dressing to reduce environmental contamination of CSS, if used in conjunction with additional antimicrobials to control organisms present in the wound.

Animals↗
PubMed1996

Hydrolysis of triclosan monophosphate by dental plaque and selected species of oral micro-organisms.

Triclosan monophosphate is a phosphorylated derivative of the antimicrobial agent, triclosan. In comparison with triclosan, it is highly soluble in aqueous solutions. It is hypothesized that, within the oral environment, triclosan monophosphate (which may be devoid of antimicrobial activity) will be hydrolyzed into triclosan by the action of microbial phosphatases. The liberated triclosan may then exert antimicrobial activity. To test this hypothesis, we designed experiments to measure the phosphatase activity of plaque and selected species of oral micro-organisms and to demonstrate hydrolysis of triclosan monophosphate. Tests comparing the minimal inhibitory concentration and minimal bactericidal concentration of triclosan and triclosan monophosphate were also undertaken. Dental plaque and the majority of the bacterial strains tested showed phosphatase activity against p-nitrophenyl phosphate which peaked below neutral pH (acid phosphatases) or above neutral pH (alkaline phosphatases). Dental plaque showed the highest levels of alkaline phosphatase (optimum at pH 9.0) and relatively high levels of acid phosphatase (optimum at pH 6.0 to 6.5). Dental plaque and selected species of micro-organisms were all capable of hydrolyzing triclosan monophosphate, albeit at different rates. The minimal inhibitory concentration and minimal bactericidal concentration values for triclosan monophosphate against eight bacterial strains were always considerably higher than the corresponding values for triclosan. Addition of triclosan monophosphate to an established culture (ca. 10(9) cfu/mL) of Capnocytophaga gingivalis growing continuously showed that triclosan monophosphate was rapidly hydrolyzed into triclosan with concomitant loss of total bacterial viability. It is therefore likely that triclosan monophosphate will be broken down into triclosan within the oral environment with concomitant antimicrobial activity.

Anti-Infective Agents, Local↗
PubMed1984

Comparison of methods to measure acute metal and organometal toxicity to natural aquatic microbial communities.

Microbial communities in water from Baltimore Harbor and from the mainstem of Chesapeake Bay were examined for sensitivity to mercuric chloride, monomethyl mercury, stannic chloride, and tributyltin chloride. Acute toxicity was determined by measuring the effects of [3H]thymidine incorporation, [14C]glutamate incorporation and respiration, and viability as compared with those of controls. Minimum inhibitory concentrations were low for all metals (monomethyl mercury, less than 0.05 microgram liter-1; mercuric chloride, less than 1 microgram liter-1; tributyltin chloride, less than 5 micrograms liter-1) except stannic chloride (5 mg liter-1). In some cases, mercuric chloride and monomethyl mercury were equally toxic at comparable concentrations. The Chesapeake Bay community appeared to be slightly more sensitive to metal stress than the Baltimore Harbor community, but this was not true for all treatments or assays. For culturable bacteria the opposite result was found. Thymidine incorporation and glutamate metabolism were much more sensitive indicators of metal toxicity than was viability. To our knowledge, this is the first use of the thymidine incorporation method for ecotoxicology studies. We found it the easiest and fastest of the three methods; it is at least equal in sensitivity to metabolic measurements, and it likely measures the effects on greater portion of the natural community.

Bacteria↗
PubMed2004

Microbial screening of UC blood units by an automated culture system: effect of delayed testing on bacterial detection.

BACKGROUND: Microbial screening is a mandatory test for banked UC blood (UCB) to comply with the code of good manufacturing practice (GMP). Cord blood banks (CBBs) are not always closely located to a GMP-licensed microbiology laboratory, resulting in time delays for transport of specimens prior to microbiological testing. This study investigated the influence of >/=24 h delays in initiating automated microbial screening on the detection of bacteria in UCB, by analysis of specimens deliberately spiked with bacteria and the recovery of bacteria from cryopreserved spiked-UCB. MATERIALS AND METHODS: UCB was processed according to standard CBB procedures and spiked with Staphylococcus epidermidis or Escherichia coli [2-2000 colony forming units (CFU)/mL]. Spiked-UCB (0.5 mL) was (1) held at room temperature (RT) and inoculated into pediatric BacT/Alert bottles (bioMérieux) at Days 1, 4 and 7 (delayed inoculation); and (2) inoculated directly (Day 0) into replicate BacT/Alert bottles and held at RT for 1, 4 or 7 days before loading onto the BacT/ALERT system (delayed loading). Spiked-UCB samples were cryopreserved. Bacterial counts were quantitated on horse blood agar plates. RESULTS: Bacterial growth in UCB spiked with a single bacterium was capable of detection by the BacT/ALERT system. S. epidermidis grew in all conditions of delayed testing (ie. delayed inoculation and delayed loading). E. coli failed to grow under conditions of delayed inoculation but grew at all time points of delayed loading. S. epidermidis and E. coli were recovered from cryopreserved spiked-UCB. DISCUSSION: Inoculation of culture bottles as soon as possible after sample preparation is preferable. Bacteria can maintain viability in BacT/ALERT bottles inoculated and held at RT for up to 7 days prior to automated culture testing. Bacteria can be successfully recovered from cryopreserved UCB.

Bacteria↗
PubMed2001

Coupling effects of osmotic pressure and temperature on the viability of Saccharomyces cerevisiae.

The osmotic tolerance of cells of Saccharomyces cerevisiae as a function of glycerol concentration and temperature has been investigated. Results show that under isothermal conditions (25 degrees C) cells are resistant (94% viability) to hyperosmotic treatment at 49.2 MPa. A thigher osmotic pressure, cell viability decreases to 25% at 99 MPa. Yeast resistance to high osmotic stress (99 Mpa) is enhanced at low temperatures (5-11 degrees C). Therefore, the temperature at which hyperosmotic pressure is achieved greatly affects cell viability. These results suggest that temperature control is a suitable means of enhancing cell survival in response to osmotic dehydration.

Cold Temperature↗
PubMed1990

Effects of paraquat on cultures of Escherichia coli: turbidity versus enumeration.

The dioxygen-dependent toxicity of paraquat has been studied both in terms of its effects on growth, monitored as increases in turbidity, and on viability, measured by plating and counting of colonies. In the absence of paraquat, turbidity and enumeration increased in parallel. However, in the presence of paraquat, turbidity increased for several hours even while enumeration indicated a marked decrease in viability. The basis for this apparent discrepancy is continued increase in size of individual cells, which have stopped dividing and are losing viability under the influence of paraquat. It can evidently be misleading to study the effects of paraquat on microorganisms in terms of changes in turbidity.

Colony Count, Microbial↗
PubMed2006

Synthesis, characterization and in vitro biological studies of novel cyano derivatives of N-alkyl and N-aryl piperazine.

Cyano derivatives of N-alkyl and N-aryl piperazine have been synthesized and screened for antibacterial and antifungal activities. All the synthesized compounds showed the antibacterial activity against pathogenic strains of Staphylococcus aureus (MTCCB 737), Pseudomonas aeruginosa (MTCCB 741), Streptomyces epidermidis (MTCCB 1824) and Escherichia coli (MTCCB 1652) and antifungal activity against pathogenic strains of Aspergillus fumigatus (ITCC 4517), Aspergillus flavus (ITCC 5192) and Aspergillus niger (ITCC 5405). All compounds showed mild to moderate antimicrobial activity. However, compounds 3c, 4a and 6 showed potent antibacterial activity against pathogenic strains used in the study. Compounds 3a, 3b, 4b, and 4d showed mild to moderate antifungal activity against Aspergillus pathogenic strains. The compounds reported in this study were assessed for there cytotoxicity using MTT colorimetric assay on Hela cells. All the compounds showed cell viability more than the control drug gentamicin, with compound 2 having highest i.e. 95% cell viability.

Acetonitriles↗
PubMed2004

Effects of several environmental factors on the anti-Listeria monocytogenes activity of an essential oil of Picea excelsa.

The effects of several environmental factors on the anti-Listeria monocytogenes activity of an essential oil of Picea excelsa were explored by determination of active concentrations using two methods and with survival curves. In trial conditions, the serovars 1/2c and 4b behaved similarly. A dose of 0.2-0.3% (v/v) of essential oil was bactericidal for 10(5)-10(7) cells contained in 1 ml of Tryptone Soy Broth Yeast Extract at pH 6 and 7 incubated at 13 and 37 degrees C and of medium supplemented with levan. Introduction of sodium caseinate, agar or fat into the test medium and use of a cheese medium decreased the bactericidal effects of the essential oil. Basic pH, addition of NaCl or use of Tryptone Soy Broth and saline solution increased its antilisterial activity. Serovar 1/2c survival curves exhibited an exponential death rate followed by a tailing effect in the presence of the minimal bactericidal concentration of the essential oil. A three log10 reduction of cell viability was obtained within 100 min in Tryptone Soy Broth Yeast Extract, within longer exposure in media supplemented with NaCl or at basic pH.

Cheese↗
PubMed1995

Control-related effective regrowth time and post-antibiotic effect of meropenem on gram-negative bacteria studied by bioluminescence and viable counts.

A study was performed to compare viable counts and bioluminescence for determining control related effective regrowth time (CERT) and postantibiotic effect (PAE) on Gram-negative bacteria after two hours of exposure to meropenem. There was a good correlation between bioluminescence and viable counts in determining the cell numbers in growing cultures of Escherichia coli. CERT was defined as the time required for the resumption of logarithmic growth and an increase of 1 log10 to occur over the pre-exposure inoculum in the test culture minus corresponding time for the control culture. PAE and CERT were studied on reference strains of Enterobacter cloacae, E. coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. At 4 x MIC of meropenem the CERTs of these four Gram-negative strains were 4.1, 4.9, 4.2, and 3.6 h, respectively, when assayed by bioluminescence. Corresponding CERTs using viable counts were 4.2, 5.0, 5.1 and 3.8 h, respectively. In contrast to this good agreement between the methods in assessing CERT, the corresponding PAEs were highly method dependent. At 4 x MIC of meropenem the PAEs on E. cloacae, E. coli, K. pneumoniae and P. aeruginosa were 3.9, 4.8, 4.7, and 3.5 h, respectively, when assayed by bioluminescene. However, the corresponding and simultaneously determined viable count PAEs were -0.4, 0.5, -0.1, and 0.7 h, respectively. The poor correlation between these methods in assessing the PAE is caused by greater initial decrease in viability compared with the less prominent initial change in cell density as measured by bioluminescence.(ABSTRACT TRUNCATED AT 250 WORDS)

Colony Count, Microbial↗
PubMed2003

Inactivation of vegetative cells, but not spores, of Bacillus anthracis, B. cereus, and B. subtilis on stainless steel surfaces coated with an antimicrobial silver- and zinc-containing zeolite formulation.

Stainless steel surfaces coated with paints containing a silver- and zinc-containing zeolite (AgION antimicrobial) were assayed in comparison to uncoated stainless steel for antimicrobial activity against vegetative cells and spores of three Bacillus species, namely, B. anthracis Sterne, B. cereus T, and B. subtilis 168. Under the test conditions (25 degrees C and 80% relative humidity), the zeolite coating produced approximately 3 log(10) inactivation of vegetative cells within a 5- to 24-h period, but viability of spores of the three species was not significantly affected.

Anti-Bacterial Agents↗
PubMed1994

The iron superoxide dismutase of Legionella pneumophila is essential for viability.

Legionella pneumophila, the causative agent of Legionnaires' disease, contains two superoxide dismutases (SODs), a cytoplasmic iron enzyme (FeSOD) and a periplasmic copper-zinc SOD. To study the role of the FeSOD in L. pneumophila, the cloned FeSOD gene (sodB) was inactivated with Tn903dIIlacZ, forming a sodB::lacZ gene fusion. By using this fusion, expression of sodB was shown to be unaffected by a variety of conditions, including several that influence sod expression in Escherichia coli: aeration, oxidants, the redox cycling compound paraquat, manipulation of iron levels in the medium, and the stage of growth. A reproducible twofold decrease in sodB expression was found during growth on agar medium containing charcoal, a potential scavenger of oxyradicals, in comparison with growth on the same medium without charcoal. No induction was seen during growth in human macrophages. Additional copies of sodB+ in trans increased resistance to paraquat. Construction of a sodB mutant was attempted by allelic exchange of the sodB::lacZ fusion with the chromosomal copy of sodB. The mutant could not be isolated, and the allelic exchange was possible only if wild-type sodB was present in trans. These results indicate that the periplasmic copper-zinc SOD cannot replace the FeSOD. The data strongly suggest that sodB is an essential gene and that FeSOD is required for the viability of L. pneumophila. In contrast, Sod- mutants of E. coli and Streptococcus mutans grow aerobically and SOD is not required for viability in these species.

Aconitate Hydratase↗
PubMed1971

Replication and recombination in ligase-deficient rII bacteriophage T4D.

Deoxyribonucleic acid replication and genetic recombination were investigated after infection of Escherichia coli with ligase-deficient rII bacteriophage T4D. The major observations are: (i) deoxyribonucleic acid synthesis is discontinuous, (ii) the discontinuities are more slowly repaired than in wild-type infection, (iii) host ligase is required for viability, and (iv) genetic recombination is increased.

Centrifugation, Density Gradient↗
PubMed1985

Bactericidal activity and induction of cell volume alterations of cephalosporins in Escherichia coli.

The bactericidal efficacy of cefuroxime and cephacetril on Escherichia coli cultures was measured by killing curves. Simultaneously bacterial cell volumes were analysed by electronic particle counting using a Coulter Counter Channelanalyser system in order to study the relationship between bactericidal activity and bacterial cell volume alterations. Various concentrations (2-120 mg/l cefuroxime and 16-120 mg/l cephacetril) and different exposure times (over a time period of 12 h) were used. Growth medium was human plasma ultrafiltrate. The bactericidal activity of cefuroxime, as measured by the rate of killing of the E. coli culture, was independent of the concentration and constant in the range 4-120 mg/l. The characteristic cefuroxime-induced change in bacterial cell volume was a marked volume increase up to a maximum of 5-fold after 160-200 min exposure with a low-grade bacteriolysis following. The cefuroxime-induced bacterial volume changes were, in accordance with the bactericidal testing, almost independent of the concentration. In contrast, the killing curves for cephacetril strongly depended on the drug concentration. However, this effect was short-lived and regrowth of the E. coli culture followed. The typical cephacetril-induced volume distribution curves were also highly concentration-dependent. With increasing drug levels bacterial cell volume increased up to 20-fold, and regrowth of a persisting bacterial population occurred at lower antibiotic concentrations. Bacteriolysis started earlier than with cefuroxime. The relationship between loss of viability and cell volume increase was more marked with cefuroxime than with cephacetril.

Autoanalysis↗
PubMed1976

Luteotropic regulation of dispersed rat luteal cells in early pregnancy.

Viable and functional luteal cells were prepared, using a combination of hyaluronidase, collagenase, and a low concentration of trypsin in a Dulbecco's modified Eagle medium containing 0.5% bovine serum albumin and 3.3 mM Ca++, from corpora lutea taken from 2-day pregnant rats. The viability and functional capacity of the dispersed cells were evaluated by electronmicroscopy and by measuring steroidogenic capicity during perifusion. Dispersed luteal cells previously exposed in vivo to biphasic prolactin (PRL) surges were found to respond during perifusion to as little as 0.5 ng/ml LH by increased steroid secretion. The net progesterone synthesis and secretion remained elevated over a time course of 2 1/2 hours perifusion, and the magnitude of the luteotropic stimulation was dose dependent on LH. However, luteotropic stimulation of LH could not be maintained beyond 2 1/2 h without renewed (in vitro) PRL exposure. PRL by itself maintained the low initial secretion rate of progesterone but demonstrated no stimulatory effect. Different steroidogenic responses were noted during the in vitro administration of LH alone and the administration of LH plus PRL. In the former case, the decreasing rate of progesterone secretion was accompanied by an increasing 20 alpha-dihydroprogesterone secretion, suggesting that luteal 20 alpha-hydroxysteroid dehydrogenase activity was not suppressed. In the latter case, progesterone secretion was maintained and 20 alpha-dihydroprogesterone secretion fell suggesting an inhibitory action by PRL against 20 alpha-hydroxysteroid dehydrogenase activity. Dispersed luteal cells, preincubated at 36 C in medium containing only PRL, retained viability and functional capacity in response to LH-PRL stimulation for periods of time up to 48 h. Preincubation with LH alone did not prolong cell viability.

20-alpha-Dihydroprogesterone↗
PubMed1984

Evaluation of a cup scrub technique for quantification of the microbial flora on bovine skin.

A cup-scrub technique devised for sampling the human skin surface microflora was evaluated in cattle. Scrub samples from bovine skin contained clumps of squama and bacterial microcolonies which were progressively broken down by shaking. This was accelerated in the presence of ballotini beads but aggregations of bacteria were still present after prolonged agitation. Vigorous shaking, particularly with beads, decreased the viability of the bacteria and optimum viable counts were obtained after manual shaking for half a minute. Immersion in buffered detergent, wash and diluting fluids for up to 2 h promoted release of bacteria from microcolonies but decreased the viability of aerobic and anaerobic pleomorphic rods and a Bacillus strain. There was no significant effect on strains of Micrococcaceae. Prolonged exposure of bacteria from scrub samples to these fluids can thus lead to both quantitative and qualitative alterations in the counts obtained, although these effects may be masked by the continuing release of bacteria from microcolonies. The cup-scrub technique provides a convenient means of quantifying changes in the bovine skin microflora but results obtained from different studies should only be compared if closely similar techniques are used.

Animals↗
PubMed1979

Preservation of mycobacteria at -70 degrees C: survival of unfrozen suspensions in transit.

Mycobacteria stored at -70 degrees C retain 100% viability and maintain their definitive taxonomic, serologic, immunologic, and pathogenic properties. When shipped at ambient temperatures, however, suspensions of all mycobacteria lose viability in transit, with those species having a narrow temperature range for growth (Mycobacterium tuberculosis and M. bovis) being most severely affected. In spite of these losses, all strains retain their definitive taxonomic properties. If care is taken in pre-testing and post-testing the microbial populations being preserved, mycobacteria are probably best shipped in the lyophilized state, and this procedure has been successfully used for several international studies.

Freezing↗