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At least 541 records · Page 30Linked to original sources
PubMed2001

The effects of freezing and thawing on the survival of Escherichia coli O157:H7 in apple juice.

Unpasteurized apple juice, adjusted to pH 3.6 to 7.0 was inoculated (10(7) CFU/ml) with single strains of E. coli O157:H7 to evaluate the effect of frozen storage on the viability of this organism. Samples were stored under frozen conditions (-20+/-2 degrees C) for up to 16 days. Cell populations were determined at regular intervals by plating onto tryptic soy agar with added pyruvate (TSAP) or onto sorbitol MacConkey agar (SMA). Populations in the neutralized juice remained unchanged during frozen storage. Populations in non-neutralized juice decreased by 1-3 log10 CFU/ml depending on the strain tested and the pH of the juice. The greatest population decrease was observed with the first freeze/thaw cycle of frozen storage (24 h) and a slow decline in survival occurred thereafter. Injury was observed after 2 weeks of storage when juice pH was at or below pH 4.2. When samples were subjected to multiple freeze/thaw cycles, loss of viability and injury increased with each freeze/thaw cycle.

Beverages↗
PubMed2002

Responses of Listeria monocytogenes to acid stress and glucose availability monitored by measurements of intracellular pH and viable counts.

Physiological aspects of the response of Listeria monocytogenes to acidic conditions and effect of glucose availability were studied by fluorescence ratio-imaging microscopy (FRIM) as compared with traditional viable counts. Three types of experiments were conducted: (i) static with measurements of intracellular pH (pHi) at extracellular pH (pHo) values ranging from pH 3.0 to 6.0 at 0.5 pH unit intervals; (ii) kinetic with monitoring of bacterial responses to changes in the pHo from the value of 6.0 to 4.0 or 3.0; (iii) survival experiments studying bacterial recovery in response to a shift to favourable conditions after a treatment at low pH. All the experiments were performed at three levels of glucose in the medium (0, 1, and 10 mM). Both survival and pHi were greatly affected by pHo and glucose availability with the highest values for CFU and pHi at highest glucose concentration and pHo values in the medium in all trials. A high correlation (R2 = 0.995) between pHi and CFU counts was observed. The pH gradient started to collapse at pHo 4 and below for trials with glucose in the medium and at pHo 5.5 and below without glucose. A recovery step was proposed after the apparently lethal treatment to assess cell viability by FRIM.

Cells, Immobilized↗
PubMed2002

Rapid viability assessment of yeast cells using vital staining with 2-NBDG, a fluorescent derivative of glucose.

A fluorescent glucose analogue, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-D-glucose (2-NBDG), which had been developed previously for the analysis of glucose uptake activity by living cells, was investigated to evaluate its applicability for assaying the viability of yeasts. Fluorescence intensities of the yeast population were measured by fluorescence spectrophotometry upon exposure to antifungal agents after staining with 2-NBDG and were compared to the number of colony forming units (CFU). A good correlation was obtained between the yeast viability, determined by the CFU, and the accumulation of 2-NBDG by yeast cells (correlation constant: r=0.98). Susceptibility testing of amphotericin B and miconazole against yeast strains by plate count and 2-NBDG fluorescence method yielded corresponding results. In conclusion, we found that staining with 2-NBDG is a rapid and sensitive method for the assessment of yeast cell viability.

4-Chloro-7-nitrobenzofurazan↗
PubMed2007

Fluorochrome and flow cytometry to monitor microorganisms in treated hospital wastewater.

Flow cytometry with a fluorescent technique (FCM/FL), epifluorescence microscopy with a fluorescent technique (EFM/FL), and a culture method were used and compared to study the microorganism population profiles in wastewater treatment. In the two non-culture methods (FCM/FL and EFM/FL), four fluorescent dyes [acridine orange (AO), 4',6-diamino-2-phenylindole dihydrochloride (DAPI), propidium iodide (PI), and YOPRO-1] were used to determine the total concentration and viability of microorganisms in the wastewater samples. Results showed that the total cell concentrations (both the bacteria and fungi) determined by using the non-culture-based methods were 18 to 67 times higher than those by the culture method (p = 0.036): the total cell concentration ranged from 1.10 x 10(7) to 2.44 x 10(8) cells/mL determined by both FCM and EFM with AO-staining method, and from 1.02 x 10(7) to 2.00 x 10(8) cells/mL by EFM with DAPI-staining method, whereas the culturable concentration of bacteria and fungi ranged from 0 to 3.22 x 10(6) CFU/mL and from 0 to 4.13 x 10(5) CFU/mL, respectively. No difference in total concentrations between dyes (AO and DAPI) and methods (FCM and EFM) were observed. By using EFM method, the microorganism viability ranged from 0.24 to 0.86 with PI staining and from 0.09 to 0.74 with YOPRO-1 staining. In the FCM analysis, the microorganism viability ranged from 0.23 to 0.87 with PI staining and from 0.18 to 0.73 with YOPRO-1 staining. In addition, the cultivability of microorganism ranged from 0 to 0.105 by the culture method. The total concentrations and viabilities of microorganisms were highly underestimated by the culture method. Results also showed that the viabilities determined by using either EFM/FL or FCM/FL were significantly higher than the cultivabilities. In addition, significant difference in viability between PI and YOPRO-1 for both EFM and FCM analysis was observed. However, the difference in viability between EFM and FCM depended on dyes. In regard to the difference between bacteria and fungi, significant difference in total concentration, viability, and cultivability was observed. In conclusion, the EFM/FL and FCM/FL methods can effectively assess total concentration and viability of microorganisms in environmental samples.

Colony Count, Microbial↗
PubMed2000

Systemic activation of coagulation and fibrynolysis in a porcine model of serogroup A streptococcal shock.

In a porcine model of Gram-positive sepsis, 28 juvenile pigs were studied to evaluate the effect of a continuous infusion of live serogroup A streptococci (GAS) on the activation of coagulation and fibrinolysis. Plasma levels of thrombin-antithrombin (TAT) complexes, tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) activities were measured using commercially available kits. The continuous infusion of GAS [(3-5) x 10(8) colony-forming units/kg per h] caused early signs of severe septicaemia in the pigs, with pulmonary hypertension, systemic hypotension, reduced cardiac output and liver hypoperfusion, ultimately leading to shock with a high mortality. There was a sequential and ordered activation of the coagulation, fibrinolytic and antifibrinolytic systems. GAS infusion induced a gradual, maximally 2.5-fold increase in plasma TAT levels. Plasma t-PA activity levels peaked at 2 h (nine-fold increase), whereas the peak of PAI-1 activity was delayed (eight-fold increase at 4 h). These findings are similar to changes observed during endotoxin infusion. This procoagulant state favours disseminated intravascular coagulation and microthrombus formation, ultimately threatening tissue viability.

Animals↗
PubMed1990

A cytoplasmically inherited mutation in the fungus Phycomyces blakesleeanus.

Fourteen mutants of the fungus Phycomyces blakesleeanus, showing high levels of resistance to copper, were isolated. In all the mutants, copper resistance behaved as a very variable and unstable trait. In the mutant strain MU102, the mutation was demonstrated to be cytoplasmically inherited. In addition, this mutant strain differed from the wild-type in growth, respiration rate, and shape and viability of spores.

Copper↗
PubMed1997

Estimation of the viability of Vibrio cholerae 0139 by assessing cell membrane integrity.

Changes in the viability of Vibrio cholerae 0139 Bengal, estimated by cellular membrane integrity, in batch culture over 35 days, were investigated. Data indicated an initial period of rapid growth with up to 30% of bacterial mortality, followed by a period of slower growth, lower culturability but higher viability, from day 7 onwards. The size of viable bacteria significantly decreased during the incubation time, whilst the size of dead bacteria showed a less pronounced decrease. V. cholerae 0139 changed from a straight or curved rod shape to a spherical shape. This study shows that BacLight dyes are a fast and useful tool to examine health risk-associated bacteria, providing useful information about their viability and concentration.

Cell Membrane↗
PubMed2005

Cell adsorption and selective desorption for separation of microbial cells by using chitosan-immobilized silica.

Cell adsorption and selective desorption for separation of microbial cells were conducted by using chitosan-immobilized silica (CIS). When chitosan was immobilized onto silica surfaces with glutaraldehyde, bacterial cells adsorbed well and retained viability. Testing of the adsorption and desorption ability of CIS using various microbes such as Escherichia coli, Aeromonas hydrophila, Pseudomonas aeruginosa, Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus, Staphylococcus epidermidis, Lactobacillus casei, Streptococcus mutans, Streptococcus sobrinus, Streptococcus salivarius, Saccharomyces cerevisiae, Saccharomyces ludwigii, and Schizosaccharomyces pombe revealed that most microbes could be adsorbed and selectively desorbed under different conditions. In particular, recovery was improved when L-cysteine was added. A mixture of two bacterial strains adsorbed onto CIS could also be successfully separated by use of specific solutions for each strain. Most of the desorbed cells were alive. Thus, quantitative and selective fractionation of cells is readily achievable by employing chitosan, a known antibacterial material.

Adsorption↗
PubMed1999

Modulation of host cell membrane fluidity: a novel mechanism for preventing bacterial adhesion.

Adhesion of bacterial enteropathogens to host mucosal surfaces is a critical primary step in the pathogenesis of diarrheal disease. We investigated the effects of altering the physical properties of eukaryotic cells on bacterial adhesion with the use of a series of three structurally dissimilar membrane fluidizers and several Escherichia coli as test strains. Lipid fluidity of the cell plasma membrane was measured by steady-state fluorescence anisotropy employing the probe 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3, 5-hexatriene. There was a dose-dependent and reversible inhibition of bacterial adhesion with increasing membrane fluidity. Time course experiments indicated that increasing membrane fluidity during the early stages of bacterial adhesion was essential for inhibition of attachment. None of the fluidizers affected the viability of either eukaryotic or prokaryotic cells. These findings demonstrate, for the first time, that changes in plasma membrane physical properties of epithelial cells can prevent microbial adhesion. This also suggests that altering the membrane properties of host cells could form a basis for novel strategies to prevent bacterial adhesion during infection in vivo.

Bacterial Adhesion↗
PubMed2002

Rational identification of new antibacterial drug targets that are essential for viability using a genomics-based approach.

In the last two decades, the search for completely novel antibacterial agents has acquired a new sense of urgency due to the remarkable rise of antibiotic resistance among key bacterial pathogens. More recently, the advent of bacterial genomics has provided investigators with the data and bioinformatic tools to rationally identify novel antibacterial targets and the genome-scaled methodologies to validate them. Only 6 years have elapsed since the publication of the first complete bacterial genome sequence, but more than 50 complete microbial genome sequences are now available. This review will discuss the advantages and limitations of the existing bacterial genome dataset for the rational identification of novel antibacterial targets. Since the ability to rapidly identify essential genes where loss of function is coincident with loss of viability is the most important task of genomics-based target validation, essentiality testing methodologies (in which molecular genetic techniques are used to determine whether or not a gene product is required for viability of the parent cell) will be surveyed and their amenability to genome-scaled analysis assessed. Finally, we will discuss the impact of bacterial genomics to date on the development of novel and effective antibiotics.

Anti-Infective Agents↗
PubMed2004

Evidence of pathogenic microbes in the International Space Station drinking water: reason for concern?

Molecular analyses were carried out on four preflight and six postflight International Space Station (ISS)-associated potable water samples at various stages of purification, storage, and transport, to ascertain their associated microbial diversities and overall microbial burdens. Following DNA extraction, PCR amplification, and molecular cloning procedures, rDNA sequences closely related to pathogenic species of Acidovorax, Afipia, Brevundimonas, Propionibacterium, Serratia, and others were recovered in varying abundance. Retrieval of sequences arising from the iodine (biocide)-reducing Delftia acidovorans in postflight waters is also of concern. Total microbial burdens of ISS potable waters were derived from data generated by an ATP-based enumeration procedure, with results ranging from 0 to 4.9 x 10(4) cells/ml. Regardless of innate biases in sample collection and analysis, such circumstantial evidence for the presence of viable, intact pathogenic cells should not be taken lightly. Implementation of new cultivation approaches and/or viability-based assays are requisite to confirm such an occurrence.

Anti-Bacterial Agents↗
PubMed1997

Room temperature storage and cryopreservation of canine platelet concentrates.

OBJECTIVE: To determine whether in vitro viability and function, and microbiological sterility, of canine platelet concentrates (PC) could be retained during storage at 20 to 24 C (room temperature [RT]) for up to 7 days and cryopreservation for 6 months. ANIMALS: 14 mature dogs. PROCEDURE: PC prepared by centrifugation of fresh blood were stored for 7 days at RT with continuous agitation. An aliquot of each was cryopreserved with 6% dimethyl sulfoxide at -74 C for 6 months. Fresh PC (day 0) were tested by microbial culture and measurement of platelet count, mean platelet volume, pH, glucose and lactate concentrations, lactate dehydrogenase activity, response to hypotonic stress, and aggregation. Tests were also performed on PC stored at RT on days 3, 5, and 7, and on the cryopreserved aliquots after thawing. RESULTS: After 7 days at RT, microbial growth was not evident, and decrease in platelet number was not significant. On the basis of pH and glucose and lactate concentrations, metabolic activity was maintained throughout RT storage. On the basis of mean platelet volume and lactate dehydrogenase activity, platelet swelling and membrane damage had occurred. Aggregatory responses decreased during RT storage, and platelets recovered poorly from hypotonic stress. After cryopreservation for 6 months, microbial growth was not evident, but platelet numbers were significantly decreased. Mean platelet volume and lactate dehydrogenase activity were significantly greater, compared with values for day-0 PC. Crypreserved platelets aggregated poorly and did not respond to hypotonic stress. CONCLUSIONS: Platelet viability and microbiological sterility are retained in canine PC stored for 7 days at RT, but platelet function pregressively decreases and day-7 platelets are substantially damaged. Crypreservation of PC results in considerable damage, compared with that of PC stored at RT. CLINICAL RELEVANCE: Similar to human PC, canine PC stored at RT for up to 5 days can be recommended for treatment.

Animals↗
PubMed1971

Enzymatic characterization of a mutant of Escherichia coli with an altered DNA ligase.

A temperature-sensitive, radiation-sensitive mutant of Escherichia coli has been assayed for DNA ligase activity in vitro. The strain contains a markedly reduced amount of DNA-joining activity, which is thermolabile. The formation of the ligase-adenylate intermediate is also temperature-sensitive in vitro. Two temperature-resistant revertants of the mutant contain normal amounts of a thermostable ligase. The mutant is killed by growth at 42 degrees C, a temperature at which it displays aberrant DNA synthesis. These results suggest that the ligase is necessary for normal DNA metabolism and viability in this strain.

Adenine Nucleotides↗
PubMed2000

Viability of bifidobacteria in commercial dairy products during refrigerated storage.

Commercial milk and two brands of yogurt containing bifidobacteria were obtained from retail outlets. All products were evaluated for viability of bifidobacteria and lactic acid bacteria during refrigerated storage at 4 degrees C. Milk was evaluated at 9, 6, and 3 days prior and past its expiration date. The yogurts were evaluated at 3, 2, and 1 week prior and past their expiration. Viability of bifidobacteria and lactic acid bacteria in milk and yogurt remained above 10(6) CFU/ml or g until the expiration date of the respective products. This microbial concentration is the recommended minimum dose to receive the health benefits of these organisms.

Animals↗
PubMed1999

Survival of Cryptosporidium parvum oocysts in source separated human urine.

The survival of Cryptosporidium parvum in source separated urine was investigated as part of a broader study on microbial risks associated with the reuse of human urine for sustainable agriculture. A dye permeability assay and in vitro excystation were the primary methods used to assess viability. In the collected urine most of the nitrogen is present as ammonia and the pH is generally around 9. Parallel investigations were made in buffers to compare possible toxic effects of urine to actual pH effects. Oocysts in the untreated urine were inactivated below the detection limit (1/300) within 63 days. This inactivation rate was significantly higher (p < 0.01) than in urine adjusted to pH 5 or 7 according to the dye permeability assay. The corresponding difference between different pH values was not seen in buffers, suggesting that the antiprotozoan effect of urine was mediated by other factors besides pH. The Swedish practice of storing urine for six months before its use thus appears satisfactory for the inactivation of Cryptosporidium oocysts.

Animals↗