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PubMed1994

Microbial contamination in four commercially available denture adhesives.

Four brands of commercially available powder denture adhesives were tested for microbial contamination. Sterile Petri plates that contained either brain-heart infusion (BHI) or Sabouraud's (SAB) agar were inoculated with 0.5 gm samples of each adhesive. The BHI plates were incubated aerobically and anaerobically at 35 degrees C and the SAB plates aerobically at 30 degrees C for 2 weeks. The sterility of these procedures was confirmed by incubating uninoculated control plates. All bottles of powder denture adhesive tested contained microorganisms. Both bacterial and fungal growth were noted. The containers of adhesive were then subjected to 10 minutes of microwave energy (650 W) to sterilize the adhesive material. After retesting, any bottles still contaminated were subjected to additional 10 minutes of microwave exposure and then retested. Seven of 24 bottles still showed evidence of microbial contamination after exposure to microwave energy. Of the seven bottles, five were of one brand. A Fisher's exact test showed significant differences in the percentage of contaminated samples between that and the other brands (p < 0.05). Microwave energy reduced microbial contamination in all samples but failed to completely sterilize seven of 24 samples.

Adhesives↗
PubMed1996

The influence of abutment surface roughness on plaque accumulation and peri-implant mucositis.

Bacterial adhesion to intra-oral, hard surfaces is firmly influenced by the surface roughness to these structures. Previous studies showed a remarkable higher subgingival bacterial load on rough surfaces when compared to smooth sites. More recently, the additional effect of a further smoothening of intra-oral hard surfaces on clinical and microbiological parameters was examined in a short-term experiment. The results indicated that a reduction in surface roughness below R(a) = 0.2 microns, the so-called "thresholds R(a)", had no further effect on the quantitative/qualitative microbiological adhesion or colonisation, neither supra- nor subgingivally. This study aims to examine the long-term effects of smoothening intra-oral hard transgingival surfaces. In 6 patients expecting an overdenture in the lower jaw, supported by endosseus titanium implants, 2 different abutments (transmucosal part of the implant): a standard machined titanium (R(a) = 0.2 microns) and one highly polished and made of a ceramic material (R(a) = 0.06 microns) were randomly installed. After 3 months of intra-oral exposure, supra- and subgingival plaque samples from both abutments were compared with each other by means of differential phase-contrast microscopy (DPCM). Clinical periodontal parameters (probing depth, gingival recession, bleeding upon probing and Periotest-value) were recorded around each abutment. After 12 months, the supra- and subgingival samples were additionally cultured in aerobic, CO2-enriched and anaerobic conditions. The same clinical parameters as at the 3-month interval were recorded after 12 months. At 3 months, spirochetes and motile organisms were only detected subgingivally around the titanium abutments. After 12 months, however, both abutment-types harboured equal proportions of spirochetes and motile organisms, both supra- and subgingivally. The microbial culturing (month 12) failed to detect large inter-abutment differences. The differences in number of colony- forming units (aerobic and anaerobic) were within one division of a logarithmic scale. The aerobic culture data showed a higher proportion of Gram-negative organisms in the subgingival flora of the rougher abutments. From the group of potentially "pathogenic" bacteria, only Prevotella intermedia and Fusobacterium nucleatum were detected for anaerobic culturing and again the inter-abutment differences were negligible. Clinically, the smoothest abutment showed a slightly higher increase in probing depth between months 3 and 12, and more bleeding on probing. The present results confirm the findings of our previous short-term study, indicating that a further reduction of the surface roughness, below a certain "threshold R(a)" (0.2 microns), has no major impact on the supra- and subgingival microbial composition.

Adult↗
PubMed2005

Selected culturable enteric bacterial populations are modified by diet acidification and the growth promotant Tylosin.

AIMS: To determine the effect of diet acidification and an in-feed antibiotic growth promotant (Tylosin, Ty) on selected culturable bacterial populations in the gastrointestinal tract (GIT) of mice. METHODS AND RESULTS: Female C57Bl mice were given a standard diet supplemented with Acid Pak (AP) or Ty in the drinking water. After 21 days, lumen and adherent populations of Enterobacteriaceae, enterococci/streptococci, and lactic acid bacteria (LAB) from the ileum, caecum, colon and faeces were enumerated. General intestinal health was assessed by the frequency of haemolytic bacteria in the different intestinal compartments. Contrary to expectations, AP and Ty significantly increased haemolytic bacteria in the lumen of the caecum and colon (P<0.05). The small but significant growth-enhancing effect of Ty (P<0.05) was associated with decreases in enterococci/streptococci and surprisingly, LAB, as well as increases in coliforms. AP, which failed to improve growth rates, reduced coliforms, had limited effects on enterococci/streptococci, and specifically failed to promote the growth of LAB populations in all intestinal compartments. Ty supplementation was also associated with a significant increase in macrolide-resistant enterococci throughout the GIT. CONCLUSIONS: Dietary acidification is less effective than Ty in modulating the population dynamics of selected culturable populations of enteric bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: The mouse can provide a useful experimental model to examine the effects of new dietary supplements, formulations or regimes on changes in microbial population dynamics, including monitoring for antibiotic resistance.

Animals↗
PubMed1994

Effect of local metronidazole application on periodontal healing following guided tissue regeneration. Microbiological findings.

The bacteriological colonization of healing periodontal defects was investigated after treatment with guided tissue regeneration using expanded polytetrafluoroethylene membranes together with local metronidazole gel (25%, 250 mg/g). Twelve patients, each with one pair of comparable defects, had the test defect treated with the membrane plus metronidazole gel and the control defect treated with the membrane alone. Thirty weeks after removal of the membrane, the median gain in probing attachment level as a percentage of the initial defect depth was 92% for the test defects and 50% for the control defects (P = 0.001). The median number of cultivable bacteria decreased from 1.2 x 10(6) at the presurgical examination to 3.0 x 10(5) at the one week examination in the test group (P = 0.02), whereas an increase was observed in the control group. Similarly, a lower median proportion of black-pigmented Gram-negative anaerobic rods was observed one week postsurgically in the test group (0.004%) compared to the control group (3.5%) (P = 0.02). Two weeks after membrane insertion, and at all following examinations, no microbiological differences between test and control group were observed. Consequently, the influence of the metronidazole gel on the treatment result appears to have been confined to the initial regeneration phase. Despite the good clinical results in the test group, all membranes from both test and control pockets were heavily colonized with bacteria at the time of removal. To ensure maximal periodontal regeneration with formation of bone, future research in this area should concentrate on reducing the microbial colonization of the wound area.

Administration, Topical↗
PubMed2001

[Study on the intestinal microbial colonization resistance in patients with chronic severe hepatitis].

OBJECTIVE: To investigate the changes of microbial colonization resistance of the bowel in patients with chronic severe hepatitis. METHODS: The fecal flora bifidobacteria, enterobacteriaceae, enterococci, and yeasts of subjects (chronic severe hepatitis, chronic hepatitis, and healthy volunteers) were quantitatively and qualitatively analyzed by Mitsuoka's methods. While the ratio of bifidobacterium to enterobacteriaceae ( B/E ratio ) was calculated. RESULTS: The number of bifidobacteria and the B/E ratio in chronic severe hepatitis group was significantly deceased compared with that of the control group (P<0.001). The number of enterobacteriaceae and yeasts was significantly increased compared with that of control group (P<0.001 and P<0.01, respectively). CONCLUSIONS: The results suggest that the intestinal microbial colonization resistance in patients with chronic severe hepatitis is seriously impaired.

Bifidobacterium↗
PubMed1999

Depth penetration and detection of pH gradients in biofilms by two-photon excitation microscopy.

Deep microbial biofilms are a major problem in many industrial, environmental, and medical settings. Novel approaches are needed to understand the structure and metabolism of these biofilms. Two-photon excitation microscopy (TPE) and conventional confocal laser scanning microscopy (CLSM) were compared quantitatively for the ability to visualize bacteria within deep in vitro biofilms. pH gradients within these biofilms were determined by fluorescence lifetime imaging, together with TPE. A constant-depth film fermentor (CDFF) was inoculated for 8 h at 50 ml. h(-1) with a defined mixed culture of 10 species of bacteria grown in continuous culture. Biofilms of fixed depths were developed in the CDFF for 10 or 11 days. The microbial compositions of the biofilms were determined by using viable counts on selective and nonselective agar media; diverse mixed-culture biofilms developed, including aerobic, facultative, and anaerobic species. TPE was able to record images four times deeper than CLSM. Importantly, in contrast to CLSM images, TPE images recorded deep within the biofilm showed no loss of contrast. The pH within the biofilms was measured directly by means of fluorescence lifetime imaging; the fluorescence decay of carboxyfluorescein was correlated with biofilm pH and was used to construct a calibration curve. pH gradients were detectable, in both the lateral and axial directions, in steady-state biofilms. When biofilms were overlaid with 14 mM sucrose for 1 h, distinct pH gradients developed. Microcolonies with pH values of below pH 3.0 were visible, in some cases adjacent to areas with a much higher pH (>5.0). TPE allowed resolution of images at significantly greater depths (as deep as 140 microm) than were possible with CLSM. Fluorescence lifetime imaging allowed the in situ, real-time imaging of pH and the detection of sharp gradients of pH within microbial biofilms.

Biofilms↗
PubMed1993

Yeasts and coliform bacteria of water accumulated in bromeliads of mangrove and sand dune ecosystems of southeast Brazil.

Yeasts and coliform bacteria were isolated from water that accumulated in the central cups and adjacent leaf axilae of two bromeliads, Neoregelia cruenta of a coastal sand dune and Quesnelia quesneliana of a mangrove ecosystem near the city of Rio de Janeiro, Brazil. The mean total coliform counts were above 10,000 per 100 mL for waters of both plants, but the mean fecal coliform counts were only 74 per 100 mL for Q. quesneliana and mostly undetected in water from N. cruenta. Of 90 fecal coliform isolates, 51 were typical of Escherichia coli in colony morphology and indol, methyl red, Volges-Proskauer, and citrate (IMViC) tests. Seven representatives of the typical E. coli cultures were identified as this species, but the identifications of nine other coliform bacteria were mostly dubious. The yeast community of N. cruenta was typical of plant surfaces with basidiomycetous yeasts anamorphs, and the black yeast Aureobasidium pullulans was prevalent. Quesnelia quesneliana had a substantial proportion of ascomycetous yeasts and their anamorphs, including a probable new biotype of Saccharomyces unisporus. Our results suggested that the microbial communities in bromeliad waters are typically autochtonous and not contaminants.

Brazil↗
PubMed2004

Microbial and sensory quality of commercial fresh processed red lettuce throughout the production chain and shelf life.

Red pigmented 'Lollo Rosso' lettuce was processed under usual and controlled conditions in an industrial plant. At different steps of the production chain (reception, shredding, washing, draining, rinsing, centrifugation, and packaging), microbial counts were evaluated. Following industrial practices, processed lettuce was packaged at 5 degrees C in sealed polypropylene (PP) bags with an initial atmosphere containing 3 kPa O(2) and 5 kPa CO(2). The numbers of psychrotrophic bacteria, coliform and lactic acid bacteria (LAB) were influenced by all the studied steps of the production chain of the fresh processed 'Lollo Rosso' lettuce. Shredding, rinsing and centrifugation in particular increased bacterial counts. During a storage period of 7 days at 5 degrees C, sensory attributes (general appearance, texture, aroma, translucency, initial and persistent off-odors, leaves superficial browning, leaves edges browning, and decay) as well as microbial counts (psychrotrophic and mesophilic bacteria, coliforms and lactic acid bacteria) were monitored. Due to high microbial counts and off-odors evaluation, a shelf life shorter than 7 days should be considered for fresh processed 'Lollo Rosso' lettuce.

Bacteria↗
PubMed2004

Evaluation of biogenic amines and microbial counts throughout the ripening of goat cheeses from pasteurized and raw milk.

The effect of the hygienic quality of milk on changes in microbial counts and biogenic amine content was evaluated during ripening of goat cheeses manufactured from pasteurized and raw milks at 1, 14, 30, 60 and 90 d. The original milk, rennet, curd and whey were also included in the study. The pH, salt content and extent of proteolysis in the cheese were also evaluated. Spermidine and spermine were the main amines in raw milk, while they were minor amines in cheeses. Other amines increased markedly during ripening, tyramine being the main amine in cheese made from raw milk and cadaverine and putrescine in those produced from pasteurized milk. Enterobacteriaceae counts decreased during ripening whereas those of lactic acid bacteria increased, especially lactobacilli and enterococci. Cheese made from raw milk showed higher microbial counts during ripening than those made from pasteurized milk, especially for Enterobacteriaceae and enterococci, counts being 2 or 3 log units higher. Raw milk cheese showed remarkably higher biogenic amines compared with pasteurized milk cheeses. Therefore, pasteurization of milk causes a decrease in final biogenic amine content of cheese as a result of the reduction of its microbial counts.

Animals↗
PubMed1998

Intra-individual variations of salivary microbial levels in young adults.

The variations of salivary levels of total viable flora, oral streptococci, Streptococcus mutans, Lactobacillus and Candida in twenty-three subjects, over a 6-month period, trying to control many potential sources of variability, were investigated. Most subjects showed log count variations of the microbial parameters larger than 1. They were 95.6% (total flora), 91.2% (oral streptococci), 73.9% (lactobacilli), 60.8% (Strep. mutans) and 30.3% (Candida). The intra-individual fluctuations were larger than the fluctuations of the means of the counts between samples. This suggests that the shifts in levels of these micro-organisms were more frequent intra-individually than at a population level. Strep. mutans levels significantly explained Streptococcus-Oral (inverse correlation), Lactobacillus and Candida variability. The reported variations were due to technical and biological factors and suggest that, in order to assess the salivary level of some microbial parameters in an individual, the mean count and the range of counts, obtained by more than two samples taken under the same conditions, should be considered.

Adult↗
PubMed2003

Microbial communities associated with geological horizons in coastal subseafloor sediments from the sea of okhotsk.

Microbial communities from a subseafloor sediment core from the southwestern Sea of Okhotsk were evaluated by performing both cultivation-dependent and cultivation-independent (molecular) analyses. The core, which extended 58.1 m below the seafloor, was composed of pelagic clays with several volcanic ash layers containing fine pumice grains. Direct cell counting and quantitative PCR analysis of archaeal and bacterial 16S rRNA gene fragments indicated that the bacterial populations in the ash layers were approximately 2 to 10 times larger than those in the clays. Partial sequences of 1,210 rRNA gene clones revealed that there were qualitative differences in the microbial communities from the two different types of layers. Two phylogenetically distinct archaeal assemblages in the Crenarchaeota, the miscellaneous crenarchaeotic group and the deep-sea archaeal group, were the most predominant archaeal 16S rRNA gene components in the ash layers and the pelagic clays, respectively. Clones of 16S rRNA gene sequences from members of the gamma subclass of the class Proteobacteria dominated the ash layers, whereas sequences from members of the candidate division OP9 and the green nonsulfur bacteria dominated the pelagic clay environments. Molecular (16S rRNA gene sequence) analysis of 181 isolated colonies revealed that there was regional proliferation of viable heterotrophic mesophiles in the volcanic ash layers, along with some gram-positive bacteria and actinobacteria. The porous ash layers, which ranged in age from tens of thousands of years to hundreds of thousands of years, thus appear to be discrete microbial habitats within the coastal subseafloor clay sediment, which are capable of harboring microbial communities that are very distinct from the communities in the more abundant pelagic clays.

Colony Count, Microbial↗
PubMed2002

Microbial contamination in inoculated shell eggs: I. Effects of layer strain and hen age.

Three Ottawa control strains and a current commercial laying stock were reared and housed under identical environmental and management conditions. Eggs were collected from each strain when hens were 32, 45, 58, 71, and 84 wk of age. The eggs were inoculated with Salmonella enteritidis (SE), Pseudomonasfluorescens (PF), or a combination of the two. After storage at 26 C, bacterial counts were obtained from the exterior shell surfaces (rinse), air cell, egg contents, and shell structure. SE and PF survived at different rates on the shell surface with as much as a 1 log difference during a given collection period. Egg content counts tended to be higher than eggshell counts in PF, whereas the opposite was true for SE. These data suggest that PF is a primary invader of eggs that is more capable of contaminating egg contents through the shell membranes than SE. The PF and SE data suggest that bacterial contamination of air cells, shells, and egg contents is more easily achieved in eggs from older hens than from younger hens. There were also differences between the strains. Control Strain 10 consistently maintained a lower level of contamination for both organisms in each sampling location. The overall results of this study suggest that genetic selection has altered the ability of eggs to resist microbial contamination and that screening for microbial integrity should be considered in the selection process among the laying egg breeders.

Age Factors↗
PubMed2002

Dietary influence of kefir on microbial activities in the mouse bowel.

AIMS: In this work the microflora present in kefir, a fermented milk product, was studied together with the effect of kefir administration on different groups of indigenous bacteria of mouse bowel. METHODS AND RESULTS: Kefir microflora was composed of lactic acid bacteria, acetic acid bacteria and yeasts. Yeast population was composed of Saccharomyces cerevisiae, S. unisporus, Candida kefir, Kluyveromyces marxianus and K. lactis. The streptococci levels in kefir treated mice increased by 10-fold and the levels of sulfite-reducing clostridia decreased by 100-fold. The number of lactic acid bacteria increased significantly. CONCLUSIONS: The administration of kefir significantly increased the lactic acid bacteria counts in the mucosa of the bowel. Ingestion of kefir specifically lowered microbial populations of Enterobacteriaceae and clostridia. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first long-term study about the effects of the kefir administration on the intestinal microflora of mice.

Animals↗
PubMed2006

Continuous flow nonthermal CO2 processing: the lethal effects of subcritical and supercritical CO2 on total microbial populations and bacterial spores in raw milk.

The effect of pressurized (<50 MPa) CO2 as a nonthermal process for bacterial reduction in raw skim milk was examined using a unique pressurized continuous flow system. The lethal effects of subcritical and super-critical CO2 applied at different temperatures and pressures toward total native psychrotrophic microbial populations, total inoculated Pseudomonas fluorescens, and total inoculated spore populations were studied and compared. Pressures between 10.3 and 48.3 MPa; temperatures of 15, 30, 35, and 40 degrees C; and CO2 concentrations of 0, 3, 66, and 132 g/kg of milk were studied. For both native populations and inoculated P. fluorescens, greater total microbial lethality was observed under supercritical CO2 conditions than under subcritical CO2 conditions. At 30 degrees C, there was no effect on total microbial lethality of increasing pressure up to 20.7 MPa with either 66 or 132 g/kg of CO2; at 35 degrees C, there was a positive relationship between pressure and lethality at CO2 levels of 132 g/kg, but no relationship at 66 g/kg of CO2. For total microbial populations and P. fluorescens, CO2 applied at 132 g/kg at 30 degrees C and pressures of 10.3 to 20.7 MPa resulted in an average standard plate count reduction of 3.81 and 2.93 log, respectively; at 35 degrees C and 20.7 MPa, maximum reductions achieved were 5.36 and 5.02 log, respectively. For both total microbial populations and inoculated P. fluorescens, CO2 exhibited a greater overall lethal effect at 132 g/kg than at 66 g/kg and a greater effect at 35 degrees C than at 30 degrees C. At 24.1 and 48.3 MPa and 40 degrees C, microbial lethality in raw aged milk treated with 3 g/kg of CO2 was not significantly different than that observed for uncarbonated milk; lethality achieved in milk treated with 132 g/kg of CO2 was significantly higher than that achieved in these 2 low-level CO2 treatments. No treatment studied had any significant impact on spore populations. Our work shows that, using the studied system, pressurized CO2 results in greater microbial lethality in milk above critical temperatures than below and suggests that a critical concentration threshold level of CO2 is required for lethal effects. Our work also suggests that supercritical CO2 processing in a continuous flow system can achieve reductions in some microbial populations equal to or better than that typically achieved during high-temperature, short-time pasteurization.

Animals↗
PubMed2005

ATP bioluminescence assay for estimation of microbial populations of fresh-cut melon.

Estimation of microbial numbers in foods by conventional microbiological techniques takes days, so there is a need for faster methods that can give results in minutes. Research was undertaken to investigate the use of bioluminescent ATP determination and a firefly luciferase assay to estimate the initial population of aerobic mesophilic bacteria on fresh-cut melons immediately after preparation and during storage at 5 or 15 degrees C for up to 12 days. Populations of aerobic mesophilic bacteria on fresh-cut cantaloupe prepared immediately from unsanitized whole melons averaged 3.42 log CFU/g, corresponding to an ATP value of 5.40 log fg/g. Populations for fresh-cut honeydew prepared from unsanitized whole melon averaged 1.97 log CFU/g, corresponding an ATP value of 3.94 log fg/g. Fresh-cut pieces prepared from cantaloupe or honeydew melons sanitized with either chlorine (200 ppm free chlorine) or hydrogen peroxide (2.5%) had similar ATP values: 3.1 log fg/g (corresponding to bacterial counts 1.7 log CFU/g) for cantaloupes and 2.6 log fg/g (corresponding to bacterial counts of 0.48 CFU/g) for fresh-cut honeydew. Positive linear correlations for ATP concentrations and microbial populations were found for fresh-cut cantaloupe (R2 = 0.99) and honeydew R2 = 0.95) during storage at 5 degrees C for up to 12 days. ATP values in fresh-cut melons inoculated with either aerobic mesophilic bacteria or yeast and mold were significantly higher (P < 0.05) than control values and parallel total plate counts on plate count agar. Results of this study indicate that the bioluminescent ATP assay can be used to monitor total microbial populations on fresh-cut melon after preparation and during storage for quality control purposes to establish specific sell-by or consume-by dates.

Adenosine Triphosphate↗
PubMed1991

Bacteriological safety of cook-chill food at the Royal Free Hospital, with particular reference to Listeria.

Cook-chill food prepared by the Catering Department of the Royal Free Hospital was examined over an 8-month period commencing October 1989. Total aerobic viable counts (TAVCs) were performed on 200 food specimens which were also examined for Listeria using selective enrichment culture. Ten of 200 food samples tested had TAVCs greater than 10(5) colony forming units (cfu) g-1, and nine of these were from non-vegetarian (meat) dishes. Although most of the food prepared by cook-chill had microbial loads within the limits recommended by current Department of Health guidelines (i.e. less than 10(5) cfu g-1), our findings for the non-vegetarian dishes suggest that extra caution is required when preparing such food. Listeria were not isolated from any food sample. In a survey of listeria faecal carriage, Listeria monocytogenes was isolated from only two of 100 faeces specimens obtained from patients.

Colony Count, Microbial↗