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Effect of acetic acid present in bagasse hydrolysate on the activities of xylose reductase and xylitol dehydrogenase in Candida guilliermondii.

The first two steps in xylose metabolism are catalyzed by NAD(P)H-dependent xylose reductase (XR) (EC 1.1.1.21) and NAD(P)-dependent xylitol dehydrogenase (XDH) (EC 1.1.1.9), which lead to xylose-->xylitol-->xylulose conversion. Xylitol has high commercial value, due to its sweetening and anticariogenic properties, as well as several clinical applications. The acid hydrolysis of sugarcane bagasse allows the separation of a xylose-rich hemicellulosic fraction that can be used as a substrate for Candida guilliermondii to produce xylitol. However, the hydrolysate contains acetic acid, an inhibitor of microbial metabolism. In this study, the effect of acetic acid on the activities of XR and XDH and on xylitol formation by C. guilliermondii were studied. For this purpose, fermentations were carried out in bagasse hydrolysate and in synthetic medium. The activities of XR and XDH were higher in the medium containing acetic acid than in control medium. Moreover, none of the fermentative parameters were significantly altered during cell culture. It was concluded that acetic acid does not interfere with xylitol formation since the increase in XR activity is proportional to XDH activity, leading to a greater production of xylitol and its subsequent conversion to xylulose.

Acetic Acid↗

Archwire ligation techniques, microbial colonization, and periodontal status in orthodontically treated patients.

Fixed or removable orthodontic appliances impede the maintenance of oral hygiene and result in plaque accumulation. Plaque retention surrounding orthodontic appliances leads to enamel demineralization caused by organic acids produced by bacteria in the dental plaque. Many studies have evaluated the effects of fixed orthodontic appliances on microbial flora and periodontal status, but only a few have evaluated the method of ligation as an additional factor. The aim of this study was to determine the changes in microbial flora and periodontal status after orthodontic bonding and to determine whether two different archwire ligation techniques affect these changes. A total of 21 orthodontic patients scheduled for fixed orthodontic treatment were selected for this split-mouth study. Two commonly used auxiliaries (elastomeric rings and ligature wires) for tying archwires were tested. Microbial and periodontal records were obtained before bonding (T0), one week later (T1), and five weeks after bonding (T2). Paired t-test and Wilcoxon signed rank test were used to compare the groups statistically. Although, teeth ligated with elastomeric rings exhibited slightly greater numbers of microorganisms than teeth ligated with steel ligature wires, the differences were not statistically significant and could be ignored. The two archwire ligation techniques showed no significant differences in the gingival index, bonded bracket plaque index, or pocket depths of the bonded teeth. However, teeth ligated with elastomeric rings were more prone to bleeding. Therefore, elastomeric ring use is not recommended in patients with poor oral hygiene.

Adolescent↗

Repeated ruminal dosing of Ruminococcus spp. does not result in persistence, but changes in other microbial populations occur that can be measured with quantitative 16S-rRNA-based probes.

Digestibility of fibre in ruminants may be improved by the introduction of highly fibrolytic strains of ruminal bacteria. This approach may be feasible if, for example, strains of Ruminococcus that are significantly more fibrolytic than the normal population of Ruminococcus are used for inoculation purposes. Introduced strains of bacteria, irrespective of ecosystem, often decline after inoculation, and in this study, highly fibrolytic strains of Ruminococcus were continuously dosed to ensure that measurements of fibre digestion were made in the presence of significant numbers of the introduced bacteria. During dosing the total culturable count increased significantly (P<0.05), but declined post-dosing. The level of dosed Ruminococcus, and total Ruminococcus, Fibrobacter succinogenes and eukaryotes measured by 16S rRNA probes increased significantly (P<0.05) during the dosing period, but also declined post-dosing. When in vitro nylon bag digestibility, feed intake or whole-tract digestibility was measured, no improvement could be measured.

Animals↗

Pacifiers: a microbial reservoir.

The permanent contact between the nipple part of pacifiers and the oral microflora offers ideal conditions for the development of biofilms. This study assessed the microbial contamination on the surface of 25 used pacifier nipples provided by day-care centers. Nine were made of silicone and 16 were made of latex. The biofilm was quantified using direct staining and microscopic observations followed by scraping and microorganism counting. The presence of a biofilm was confirmed on 80% of the pacifier nipples studied. This biofilm was mature for 36% of them. Latex pacifier nipples were more contaminated than silicone ones. The two main genera isolated were Staphylococcus and Candida. Our results confirm that nipples can be seen as potential reservoirs of infections. However, pacifiers do have some advantages; in particular, the potential protection they afford against sudden infant death syndrome. Strict rules of hygiene and an efficient antibiofilm cleaning protocol should be established to answer the worries of parents concerning the safety of pacifiers.

Analysis of Variance↗

The influence of the growth phase of enteric bacteria on electrotransformation with plasmid DNA.

Salmonella typhimurium LB5000 and Escherichia coli JM109 were transformed by electroporation. In accordance with the chemical transformation methods, the growth phase of these electrocompetent bacteria had a strong impact on transformation efficiency. Survival of bacteria after the high-voltage electrical pulse was also influenced by the growth phase. Both bacterial species were most successfully electrotransformed when microbial cells were harvested at the late lag phase. The second optimum for transformation reached E. coli cells in the mid-exponential and S. typhimurium cells in the late exponential phase. Transformation efficiencies ranged from 3.4 x 10(4) to 2.7 x 10(5) transformants per microgram DNA in the case of S. typhimurium and from 2.8 x 10(2) to 8.8 x 10(5) transformants per microgram DNA in the case of E. coli. Survival of cells after the electrical pulse in late lag and late exponential phases was about 20% higher than during other phases of growth. Preparing electrocompetent cells from later phases of their growth is more useful for practice, because it provides more biomass with good yield of transformants.

Colony Count, Microbial↗

Influence of a synbiotic mixture consisting of Lactobacillus acidophilus 74-2 and a fructooligosaccharide preparation on the microbial ecology sustained in a simulation of the human intestinal microbial ecosystem (SHIME reactor).

Lactobacillus acidophilus 74-2, which is used in probiotic products, was administered, with fructo-oligosaccharide in a milk-based product, to the second vessel (duodenum/jejunum) of the SHIME reactor, an in vitro simulation of the human intestinal microbial ecology. The main focus of this study was to monitor the changes of the population density of selected bacterial species in the intestine and the changes of metabolic activities during the supplementation of L. acidophilus and fructooligosaccharide in the SHIME reactor. Interestingly, the addition of L. acidophilus 74-2 with fructooligosaccharide gave rise to an increase of bifidobacteria. Moreover, major positive changes occurred in the production of volatile fatty acids: a strong upward trend was observed especially in the case of butyric acid and propionic acid. Furthermore a noticeable increase of beta-galactosidase activity was monitored, while the activity of beta-glucuronidase, generally considered undesirable, declined.

Animals↗

Physiological function of exopolysaccharides produced by Lactococcus lactis.

The physiological function of EPS produced by Lactococcus lactis was studied by comparing the tolerance of the non-EPS producing strain L. lactis ssp. cremoris MG1614 and an EPS producing isogenic variant of this strain to several anti-microbial factors. There was no difference in the sensitivity of the strains to increased temperatures, freezing or freeze-drying and the antibiotics, penicillin and vancomycin. A model system showed that EPS production did not affect the survival of L. lactis during passage through the gastrointestinal tract although the EPS itself was not degraded during this passage. The presence of cell associated EPS and EPS in suspension resulted in an increased tolerance to copper and nisin. Furthermore, cell associated EPS also protected the bacteria against bacteriophages and the cell wall degrading enzyme lysozyme. However, it has not been possible, so far, to increase EPS production using the presence of copper, nisin, lysozyme or bacteriophages as inducing factors.

Bacteriophages↗

Chagasic megacolon and proximal jejunum microbiota.

BACKGROUND: Among other factors, control of jejunal microflora depends on intestinal emptiness, and it can be impaired by Chagas disease. This study was developed to identify the microecology of the proximal jejunum in chagasic megacolon. Our objective was to characterize both the jejunal microbial stasis before surgery and the microflora after surgical treatment in patients with chagasic megacolon. METHODS: The intestinal fluids were collected, and the proximal jejunum microflora was analyzed. RESULTS: Preoperative microflora had shown an increase in bacteria compatible with bacterial overgrowth syndrome, mainly facultative and strict anaerobes microorganisms and fungi. The microflora had changed in the postoperative period in 83% of these patients, with significant decrease in the number of transient microorganisms. CONCLUSIONS: Chagasic megacolon was related to an increase in jejunal microflora. By removing the impairment of the colon there was a transient decrease in the proximal jejunum microflora.

Adult↗

Microbiology of healthy and diseased periodontal sites in poorly controlled insulin dependent diabetics.

A group of poorly-controlled insulin dependent diabetes mellitus (IDDM) patients were examined in a cross-sectional design for total microbial levels, microbial incidence, and the percent levels of selected periodontal microorganisms. These organisms were selected on the basis of prior reports that associated them with either periodontal disease or health. One periodontally-healthy and one periodontally-diseased site were examined in each IDDM patient. Increased levels of the periodontal pathogens Prevotella intermedia, P. melaninogenica spp., Bacteroides gracilis, Eikenella corrodens, Fusobacterium nucleatum and Campylobacter rectus (formerly Wolinella recta) were found at the periodontal diseased sites. Increased prevalence of the organisms P. intermedia, P. melaninogenica spp., and C. rectus were found at the diseased sites. A significantly higher percentage of P. intermedia was found at the sites exhibiting deep pockets and attachment loss.

Adult↗

Natural milk cultures for the production of Argentinian cheeses.

Samples (32) of natural milk cultures used in the Santa Fe, Argentina, area for soft and semihard cheese production were examined. The microbial composition (including lactic acid microflora characterization) and technological parameters (acidifying and proteolytic activities) were evaluated. The cultures contained mainly thermophilic lactic acid bacteria, identified as Streptococcus salivarius subsp. thermophilus (96.8% of the total strains) and Enterococcus spp. The strains showed a low proteolytic activity. The isolates of S. salivarius subsp. thermophilus exhibited a widespread phage resistance. The nonlactic microflora comprised coliforms, yeasts, spore-forming bacteria and lactate fermentative bacteria. The samples showed an acidity level from 0.38 to 0.69% lactic acid (pH from 4.25 to 5.75). The acidifying activity was optimal at 45 degrees C. The advantages and disadvantages of the employment of natural milk starters are discussed.

Animals↗

Microbiological quality of fermented cassava flour 'kpor umilin'.

A study was conducted on the microbiological quality of traditionally fermented cassava flour 'kpor umilin' and a laboratory modification of the method developed with a view to upgrading the traditional process. Microbial analysis of samples from both processing methods was evaluated using standard microbiological methods. The bacterial count in the traditionally processed flours ranged from 2.7 x 10(3) to 1.2 x 10(7) cfu/g, while the modified sample had a count of 3.5 x 10(2) cfu/g. The predominant flora were Leuconostoc spp., Lactobacillus spp., Staphylococcus spp., Micrococcus spp., Bacillus spp., Streptococcus spp. and coliforms such as Escherichia coli. Other isolates were Enterobacter spp. and proteus spp. The sample from the modified method had a negligible (4 MPN/100 ml) coliform count. The yeast and mould count ranged from 1.9 x 10(3) cfu/g to 3.9 x 10(5) cfu/g in the traditionally processed samples as compared with 1.5 x 10(3) cfu/g in the sample from the modified process. Aspergillus spp., Penicillium spp., Saccharomyces spp. and Geotrichum candidum were the predominant fungi. The poor microbiological quality of 'kpor umilin', processed traditionally, calls for urgency in upgrading the processing and preservation methods as developed in this study as well as proper education of the local processors on good manufacturing practices.

Colony Count, Microbial↗

[Changes in the microbial ecology and morphology of large intestine biofilm in albino rats under the influence of rifampicin].

After oral administration in single doses of 40 and 160 mg/kg to Wistar rats rifampicin was detected in 3 hours in the contents of the large intestine The maximum rifampicin concentrations in the feces (120 and 300 micrograms/g) were detected in 24 hours. The antibiotic was present in the animal feces for 6-7 days. The antibiotic administration led to marked changes in microbiocenosis of the large intestine: disappearance of coccal flora, lower quantities of Escherichia and Lactobacillus and lower total quantities of anaerobic bacteria. It was also accompanied by changes in the structure of the typhlon biofilm. By the 28th day after discontinuation of the antibiotic administration (the end of the observation period) the large intestine microflora did not recover completely.

Animals↗

Significance of sinking particles in the distribution of motile Aeromonas during the winter season.

The role of sinking particles in the distribution of motile Aeromonas species was studied during the winter season. Various environmental parameters and microbial populations were investigated to elucidate the relationship with motile aeromonads. Motile Aeromonas species were enumerated by most probable number technique with alkaline peptone water as the enrichment medium and modified pril-xylose ampicillin agar as the plating medium. Aeromonas species were isolated in a water column in any one of the two procedures but sediment and plankton samples exhibited an irregular isolation pattern for these organisms. Aeoromonas species were continuously isolated in sinking particles with the highest counts during January. Of the 206 isolates identified, three known motile Aeromonas species were observed of which A. caviae accounted for 51.4% of the total. Toxin characterization showed that 20% of the strains produced haemolysin as well as cytotoxin, and A. hydrophila was highly toxigenic. Statistical analyses revealed that nutrients govern the distribution of Aeromonas. It may be that riverine discharge influences the distribution of motile aeromonads in this environment.

Aeromonas↗

The relationship between alveolar bone loss and proximal caries in children: prevalence and microbiology.

The present study describes the prevalence of alveolar bone loss (ABL) in children in relation to caries, contact and space loss. In addition, the microbial composition of the subgingival plaque of 20 sites, from 5 children, is presented. Bite-wing radiographs from 500 children were examined. ABL was evident in: 99 sites from 60 children; > 1 site in 27 children; the maxilla only in 34 children; the mandible only in 17 children; both arches in 9 children; 37 sites with no caries; 4.9 percent of all sites with proximal caries; 15.8 percent of all sites with contact loss; and 20.5 percent of all sites with mesial drift. Anaerobic bacteria were cultured from all 20 sites. No significant differences in the percentages of colony forming units of Actinobacillus actinomycetemcomitans and black pigmented Bacteroides were found among sites with/without bone loss, with/without caries or probing depths smaller/equal or larger than 2.5 mm.

Aggregatibacter actinomycetemcomitans↗

An evaluation of saline irrigation and comparison of povidone-iodine and antibiotic in the surface decontamination of donor eyes.

We evaluated the effect of saline irrigation on 38 pairs of donor eyes and determined the relative efficacy of antibiotic rinse/storage (neomycin-polymyxin B-gramicidin) and povidone-iodine immersion. Microbial growth was found in 49 (64.5%) of 76 eyes from which cultures were taken before irrigation, and only four (8.2%) became culture negative after irrigation. Of 27 eyes culture negative before irrigation, 15 (55.6%) became positive after irrigation. One eye of each pair was then assigned randomly to antibiotic rinse/storage and the other eye to 3-minute immersion in 1% povidone-iodine. Both antimicrobial treatments decreased the numbers of positive cultures, with povidone-iodine showing an advantage that did not reach significance. These results demonstrate that saline irrigation of donor eyes in situ does not reduce surface contamination and that povidone-iodine immersion should be considered as an alternative for donor eye decontamination.

Anti-Bacterial Agents↗

A novel method to control the balance of skin microflora. Part 1. Attack on biofilm of Staphylococcus aureus without antibiotics.

BACKGROUND: Staphylococcus aureus (SA) is usually present in atopic dry skin, and not only in regions seriously affected by atopic dermatitis. SA discharges various toxins and enzymes that injure the skin, and forms a biofilm from fibrin fiber and glycocalyx; the biofilm is important for adhesion of SA to the skin and for resistance to anti-microbial agents. Even highly effective moisturizers do not work perfectly on atopic dry skin. Staphylococcus epidermidis (SE) is a major constituent of skin microflora on healthy human skin, and provides protection against the growth of pathogenic bacteria. OBJECTIVES: Since treatment with anti-microbials may lead to re-growth of SA, which grows faster than other Staphylococci and often shows antibiotic resistance, we searched for novel approaches to control the skin-microfloral balance without using conventional anti-microbials. METHOD: Biofilm formation by SA in vitro was observed in detail using scanning electron microscopy. Approximately 500 substances were screened for a selective effect on SA growth and SA biofilm. RESULTS: We found that xylitol inhibited the formation of glycocalyx, and farnesol dissolved fibrin fibers. Farnesol suppressed the growth of only SA, and did not affect that of SE. Xylitol and farnesol synergistically inhibited biofilm formation by SA. CONCLUSION: Xylitol and farnesol have potential for controlling the skin-microfloral balance because of their selective effects and inhibition of biofilm formation. They might provide a useful and safe method to care for skin colonized by SA, without using antibiotics.

Administration, Cutaneous↗

Bacterial community shifts in organically perturbed sediments.

Bacterial abundance, diversity and sediment function were investigated in organically perturbed sediments under Tasmanian salmon (Salmo salar) farms and adjacent reference sites. Bacterial numbers increased as farming and organic loading progressed through the farm stocking cycle and declined during the fallow period, although not to prestocking levels. Bacterial numbers ranged between approximately 2 x 10(8) and 3 x 10(9) cells per gram of sediment and were higher at cage sites than reference sites. Microelectrode and respiration data also demonstrated a clear effect of organic loading on sediments. Denaturing gradient gel electrophoresis (DGGE) showed that bacterial communities shifted both in response to farm loading and its cessation. A seasonal effect on microbial communities was also evident. Although bacterial communities did shift again during the fallowing period, this shift was not necessarily a return to preloading communities. The complexity of community shifts may be affected by the vast functional redundancy of bacterial groups. All bacterial communities, including those at reference sites, were highly dynamic. Respiration studies of amended sediments indicated that fish farm sediments were at least as resilient and diverse as reference site communities. The results of this study indicate that the functional redundancy of highly complex bacterial communities contributes to their robustness. The relationship between diversity and stability in bacterial communities remains unclear and requires further investigation before an understanding of bacterial response to perturbation is possible.

Animals↗

Conjugal transfer of R68.45 and FP5 between Pseudomonas aeruginosa strains in a freshwater environment.

Recent concern over the release of genetically engineered organisms has resulted in a need for information about the potential for gene transfer in the environment. In this study, the conjugal transfer in Pseudomonas aeruginosa of the plasmids R68.45 and FP5 was demonstrated in the freshwater environment of Fort Loudoun Resevoir, Knoxville, Tenn. When genetically well defined plasmid donor and recipient strains were introduced into test chambers suspended in Fort Loudoun Lake, transfer of both plasmids was observed. Conjugation occurred in both the presence and absence of the natural microbial community. The number of transconjugants recovered was lower when the natural community was present. Transfer of the broad-host-range plasmid R68.45 to organisms other than the introduced recipient was not observed in these chambers but was observed in laboratory simulations when an organism isolated from lakewater was used as the recipient strain. Although the plasmids transferred in laboratory studies were genetically and physically stable, a significant number of transconjugants recovered from the field trials contained deletions and other genetic rearrangements, suggesting that factors which increase gene instability are operating in the environment. The potential for conjugal transfer of genetic material must be considered in evaluating the release of any genetically engineered microorganism into a freshwater environment.

Colony Count, Microbial↗