Change In Colony-forming Units Research Articles

Evaluation of Escherichia coli Inactivation at High Altitudes Using Solar Water Disinfection.

Solar disinfection (SODIS) is an effective method for microbiologic inactivation of contaminated water using ultraviolet rays at low elevations. The aim of this study was to determine the effectiveness of SODIS at higher elevations. The ability of SODIS to inactivate Escherichia coli bacteria was evaluated at an altitude of ≥1600 m using Nalgene bottles, disposable plastic water bottles, and Ziploc plastic bags. Bacterial viability was determined through measurement of colony forming units (CFUs). Decreases in CFUs were determined at each time point relative to those at the baseline, and a multivariable regression analysis was used to assess significant changes in CFUs. Bacterial CFUs in exposed containers decreased by >5 log after 6 h of exposure to sunlight. In contrast, the CFUs remained nearly unchanged in unexposed containers, showing a mean decrease of 0.3 log. By 2 h, bacterial inactivation at high altitudes was 1.7-fold greater than that at lower altitudes (P<0.05). By 6 h, nearly all bacteria were inactivated at high or low altitudes. At 6 h, no statistical difference was observed in the efficiency of inactivation between elevations. Compared with Nalgene bottles, plastic bottles had a 1.4-fold greater decrease in CFUs (P<0.05). No statistical difference in bacterial inactivation was found between plastic bottles and plastic bags. At high altitudes, SODIS is an effective method for inactivating E coli. Further research investigating other microorganisms is warranted to determine whether SODIS is suitable for disinfecting contaminated water at high altitudes.

In Vitro Evaluation of Bacterial Adhesion and Bacterial Viability of Streptococcus mutans, Streptococcus sanguinis, and Porphyromonas gingivalis on the Abutment Surface of Titanium and Zirconium Dental Implants.

Objective To evaluate the in vitro adherence and viability of 3 bacterial species Streptococcus mutans (ATCC 25175), Streptococcus sanguinis (ATCC 10556), and Porphyromonas gingivalis (ATCC 33277) on the surfaces of dental implants of titanium, zirconium, and their respective fixing screws. Methods Two analysis groups were formed: group 1 with 3 titanium pillars and group 2 with 3 zirconium pillars, each with their respective fixing screws. Each of these groups was included in tubes with bacterial cultures of Streptococcus mutans (ATCC 25175), Streptococcus sanguinis (ATCC 10556), and Porphyromonas gingivalis (ATCC 33277). These samples were incubated at 37°C under anaerobic conditions. Bacterial adherence was assessed by measurement of the change in colony-forming units (CFU), and bacterial viability was evaluated with the colorimetric test of 3-(4,5-dimethylthiazol-2)-2,5 diphenyl tetrazolium bromide (MTT). Results The bacterial adhesion in the titanium abutments was higher for Streptococcus mutans (190.90 CFU/mL), and the viability was greater in Porphyromonas gingivalis (73.22%). The zirconium abutment group showed the highest adherence with Streptococcus mutans (331.82 CFU/mL) and the highest bacterial viability with the S. sanguinis strain (38.42%). The titanium fixation screws showed the highest adhesion with S. sanguinis (132.5 CFU/mL) compared to the zirconium fixation screws where S. mutans had the highest adhesion (145.5 CFU/mL). The bacterial viability of S. mutans was greater both in the titanium fixation screws and in the zirconium fixation screws 78.04% and 57.38%, respectively. Conclusions Our results indicate that there is in vitro bacterial adherence and viability in both titanium abutments and zirconium abutments and fixation screws for both. Streptococcus mutans is the microorganism that shows the greatest adherence to the surfaces of both titanium and zirconium and the fixing screws of the latter. On the contrary, bacterial viability is greater on the titanium abutments with P. gingivalis than on the zirconium abutments with S. sanguinis. With respect to the fixation screws, in both cases, the viability of S. mutans was greater with respect to the other bacteria. In general, the titanium abutments showed less adherence but greater bacterial viability.

Evaluation of Cronobacter Growth and Phenotypic Variation Under Modified Culture Conditions.

Cronobacter sakazakii is an opportunistic pathogen known to cause acute meningitis and necrotizing enterocolitis in neonates and immunocompromised individuals. It has been isolated from a wide range of food and environmental samples, and has been linked to outbreaks associated with powdered infant formula. This study was carried out to assess variations in growth conditions (temperature, pH, and sugar supplement) and to establish how these changes impact phenotypic characteristics for successful recovery and identification of Cronobacter, particularly for routine surveillance purposes. A total of six Cronobacter isolates were tested to evaluate the above growth conditions, including three ATCC Cronobacter reference and three environmental isolates obtained from regulatory sample screening. Although only slight changes in colony-forming units were observed across the pH range and the sugars tested, the morphology was significantly impacted by changes in these growth factors. Incubation between 30 and 50 °C resulted in growth after 24 h, and the growth was slower at ambient temperature and colony formation was most robust at 30 °C. Results of this study suggest that 30 °C may be suitable for recovery of some Cronobacter strains, and minor variations in growth conditions can alter colony morphology and appearance. Expression of unique biological characteristics based on phenotypic observations may be beneficial for differentiating various Cronobacter strains.

PH-dependent antibacterial effects on oral microorganisms through pure PLGA implants and composites with nanosized bioactive glass

Biomaterials made of biodegradable poly(α-hydroxyesters) such as poly(lactide-co-glycolide) (PLGA) are known to decrease the pH in the vicinity of the implants. Bioactive glass (BG) is being investigated as a counteracting agent buffering the acidic degradation products. However, in dentistry the question arises whether an antibacterial effect is rather obtained from pure PLGA or from BG/PLGA composites, as BG has been proved to be antimicrobial. In the present study the antimicrobial properties of electrospun PLGA and BG45S5/PLGA fibres were investigated using human oral bacteria (specified with mass spectrometry) incubated for up to 24h. BG45S5 nanoparticles were prepared by flame spray synthesis. The change in colony-forming units (CFU) of the bacteria was correlated with the pH of the medium during incubation. The morphology and structure of the scaffolds as well as the appearance of the bacteria were followed bymicroscopy. Additionally, we studied if the presence of BG45S5 had an influence on the degradation speed of the polymer. Finally, it turned out that the pH increase induced by the presence of BG45S5 in the scaffold did not last long enough to show a reduction in CFU. On the contrary, pure PLGA demonstrated antibacterial properties that should be taken into consideration when designing biomaterials for dental applications.

Protective Effect of Polysaccharides Isolated from &lt;i&gt;Tremella fuciformis&lt;/i&gt; against Radiation-induced Damage in Mice

WTF-B, a type of water-soluble homogeneous polysaccharide, was isolated and purified from Tremella Fuciformis. To investigate the radioprotective effect of WTF-B, we employed a 30-day survival assay. Mice were treated with WTF-B once per day for three consecutive days before 8-Gy gamma irradiation. The treatment groups receiving 54 and 72 mg/kg body weight (b.w.) of WTF-B showed 50% survival post-irradiation. The hematological parameters of the peripheral blood indicated that WTF-B, when administered at doses of 72 mg/kg b.w., significantly restored hemoglobin, white blood cell counts and red blood cell counts by the 14th day and 18th day. In addition, spleen colony forming units (CFU-S), the number of nucleated cells in bone marrow (BMNC) and spleen index were used to investigate the radioprotective effect of WTF-B on the hematopoietic system. The treatment groups receiving WTF-B at 18, 54 and 72 mg/kg b.w. doses presented significantly higher BMNC compared to radiation-only group. The group administered 72 mg/kg b.w. WTF-B presented a significant change in CFU-S compared to the radiation-only group. We also completed micronucleus and chromosome aberration assays to explore genotoxicity. The results of those assays indicated that the number of micronuclei induced by 2-Gy irradiation in a group treated with 72 mg/kg b.w. WTF-B decreased from 30.30‰ to 11.32‰. The chromosomal aberration produced by 3-Gy irradiation in the group receiving 72 mg/kg b.w. WTF-B decreased from 56.01% to 28.13%. The results of the present study indicate a potential use for WTF-B as a radioprotector.

Gaseous ozone treatment inactivates Listeria innocua in vitro

To investigate the effects of ozone on inactivation of Listeria innocua on solid media. Suspensions of L. innocua ranging from 4.5 x 10(4 )- 6.4 x 10(4) CFU ml(-1) were inoculated onto potato dextrose agar (PDA, pH 5.6 and 6.8) and nutrient agar (NA, pH 6.0 and 6.8), then exposed to gaseous ozone. Variable factors included postinoculation standing time at 20 degrees C before exposure to ozone, ozone concentration, treatment duration and treatment temperature (5 or 20 degrees C). The interaction among ozone concentration, treatment duration, media and temperature in effecting changes in colony-forming units (CFU) was significant. The 100 nl l(-1) ozone treatment for 2 h reduced the microbial populations by 2-3 log CFU ml(-1). Cell viability decreased more rapidly on PDA than on NA. The average time to obtain a 2 log CFU ml(-1) reduction was 1.3 h at 20 degrees C and 2.5 h at 5 degrees C (P < 0.001). Gaseous ozone effectively inactivates L. innocua at concentrations of 50 and 100 nl l(-1) during short exposure times at both 5 and 20 degrees C. The Gompretz model can be utilized for determining the response of L. innocua to ozone over time. This study provides useful information on ozone inactivating Listeria spp., which may be imposed on ensuring quality and safety of horticultural produce and food products.

Increase in the Culturable Cell Number of Escherichia coli during Recovery from Saline Stress: Possible Implication for Resuscitation from the VBNC State.

The change in colony-forming units (CFUs) in saline-stressed Escherichia coli cultures was examined. Exposure of E. coli cells to high saline stress decreased the number of CFUs significantly. When the culture was relieved from the stressful condition, the number of CFUs returned within 2 hr to the same level as before the stress. This recovery in the number of CFUs seemed to be independent of DNA synthesis and cell division, because the same phenomenon was also observed in the presence of nalidixic acid. However, the total cell number enumerated by microscopy was the same before and after relief from saline stress. This phenomenon was considered to be the result of the resuscitation of cells that had been in the VBNC state during saline stress. Our method, in which we examined CFU recovery in the presence of a DNA synthesis inhibitor, might be useful for further VBNC study.

In vitro antimicrobial activity of the human neutrophil cytosolic S-100 protein complex, calprotectin, against Capnocytophaga sputigena.

Calprotectin is a complex of two anionic proteins found in abundance in the cytosol of neutrophils, certain macrophages, and oral epithelial keratinocytes. Bacteria of the genus Capnocytophaga are pathogens of periodontal origin which can cause systemic infection in neutropenic subjects. Recently, it has been observed that Capnocytophaga may be internalized by neutrophils within the cytosol rather than within a membrane-delimited phagosome. The purpose of this study was to test the in vitro antibacterial effect of the cytosolic complex, calprotectin, against Capnocytophaga sputigena. Calprotectin was purified from the cytosol of human neutrophils by gel filtration and anion exchange FPLC, and it exerted potent in vitro antimicrobial effects against C. sputigena. Net bacteriostatic activity was exerted up to 18 h, after which bactericidal effects were observed. Both net bacteriostatic and bactericidal activity occurred at concentrations above 20 micrograms/mL and exhibited identical dose-response characteristics. Particle counts increased in the presence of calprotectin, despite net bacteriostasis as assessed by changes in colony-forming units (CFU). Dose-response characteristics and direct particle counts suggested that net bacteriostatic effects were the result of balanced cell division and death, rather than suspension of cell division. We conclude that calprotectin can be a significant contributor to host defense against infection by Capnocytophaga.

Phagocytosis of Mycoplasma salivarium by human polymorphonuclear leukocytes and monocytes.

Mycoplasmacial activity was exhibited by human peripheal blood leukocytes in the absence of detectable specific antiserum. After incubation of varying concentrations of Mycoplasma salivarium with leukocytes, changes in colony-forming units (CFU) of this species per millilter occurred. The most noticeable decrease in CFU per milliliter was then the incubation mixtures contained five mycoplasmas per leukocyte. At this ratio, the mycoplasmacial min of incubation. Continued incubation demonstrated a tenfold decrease in CFU per milliliter by 4 h. Electron micrographs of incubated mixtures of human leukocytes and M. salivarium showed this mycoplasma to be phagocytized by monocytes and neutrophils whenever mutual contact or pseudopodial formation occurred. The process was continuous. Numerous phagocytic vacuoles developed which contained multiple ingested microorganisms. After the cytoplasmic granules of the leukocytes fused with the phagocytic vacuole, the phagocytized mycoplasmas became disrupted and unrecognizable.

Release of 51Cr-Endotoxin from Bacteria as an Assay of Serum Bactericidal Activity

Abstract In order to facilitate the study of normal serum on Gram-negative bacteria we have developed a bactericidal assay based on the release of 51Cr from labeled bacteria. Release of radioactivity was shown to be mediated by complement and paralleled the bactericidal action of sera measured by change in colony-forming units. Serum from the C6 deficient rabbits and serum that had been treated to inactivate complement did not release 51Cr. The rate of release of 51 Cr was decreased by absorption of serum with Bentonite but the maximum percentage released was not decreased. 51Cr was not released from serum resistant bacteria. The 51Cr appears to be associated with the lipopolysaccharide fraction of the bacteria. Since this structure is the target for the bactericidal reaction, the release of 51Cr is a direct measure of the effect of complement on the bacteria. Under most conditions this reaction leads to cell death so that the release of 51Cr can be used as an assay of bactericidal activity. It was shown that the assay was feasible with Escherichia coli, Salmonella and Hemophilus influenza B.

Effect of Endotoxin on the Kinetics of Hemopoietic Colony-Forming Cells in Irradiated Mice

Hemopoietic recovery in irradiated animals is hastened by the action of bacterial endotoxin given as a single injection either before or shortly after irradiation (1). Bone marrow cellularity, peripheral blood granulocyte, erythrocyte, platelet, and lymphocyte concentrations, and survival are favorably affected and, in the mouse, the number of endogenous spleen colonies is increased (2-4). The fact that endotoxin is effective when given after irradiation rules out several modes of action. These include chemical protection, hypoxia, increased numbers of stem cells at the time of irradiation, and decreased sensitivity of the stem cells to the immediate killing action of radiation. Among the possibilities remaining are recovery of cells that otherwise would die on the resumption of mitosis, early resumption of mitosis, and shortening of the cell cycle. Means of exploring such possibilities are provided by the technique for enumerating colony-forming units (CFU) developed by Till and McCulloch (5-7). Suspensions containing hemopoietic cells are injected into recipient mice whose endogenous spleen colony formation has been suppressed by lethal irradiation and the resulting spleen colonies are counted 10 days later. The transplanted colonyforming units have many of the characteristics of stem cells (8), and in the present experiments we assume that their enumeration provides an assay of stem cells. The experiments presented here concern population changes in colony-forming units in spleen and femoral marrow resulting from an injection of Salmonella typhosa endotoxin 24 hours before or a few minutes after irradiation with 400 rads.