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[A comparative study of the efficacy of the action of a number of local antimicrobial preparations on the species and quantitative composition of the microbial flora in periodontal pockets].

The effects of a number of antibiotics were studied with the use of phase contrast microscopy. The time course of the qualitative and quantitative changes of the periodontal pouch microflora evidences that a mixture of trichopol with chlorohexidine is the most effective therapy for periodontal pouch sanitization.

Anti-Infective Agents, Local↗

High-pressure destruction kinetics of Listeria monocytogenes on pork.

Packaged fresh pork chops (30-g samples) containing an indigenous bacterial population of approximately 10(7) CFU/g were inoculated with 10(7) CFU of Listeria monocytogenes Scott A per g, heat sealed, and subjected to high-pressure processing at 200 to 400 MPa for up to 90 min. Total counts and the number of surviving L. monocytogenes cells were determined by a spread plate technique on tryptic soy agar and modified Oxford medium, respectively. The pressure destruction was characterized by a dual-behavior, consisting of a step change in the number of survivors (Pk0) with the application of a pressure pulse and a first-order rate drop in the number of survivors during the pressure hold period. Higher pressures resulted in higher rates of microbial inactivation, as indicated by their associated lower D values (and higher k values). The pressure sensitivities of the kinetic parameters were evaluated on the basis of Arrhenius and pressure death time (PDT)-type models. The results suggested that L. monocytogenes was more resistant to pressure inactivation than the indigenous microflora (the volume change of activation, deltaV* [Arrhenius model]), and Zp values (PDT model) were -4.17 x 10(-5) m3 mole(-1) and 134 MPa for indigenous microflora and -3.43 x 10(-5) m3 mole(-1) and 163 MPa for L. monocytogenes respectively.

Animals↗

Effects of high and fluctuating pressure on microbial abundance and activity in a water hydraulic system.

The effects of high and fluctuating pressure up to 220 bar on microbial growth and activity were determined in a pilot-scale water hydraulic system. An increase in the pipeline pressure from 70 to 220 bar decreased the total and the viable cell number in the pressure medium from 2.2(+/-0.5)x10(5) to 4.9(+/-1.5)x10(4) cells/ml and from 5.7(+/-2.8)x10(4) to 1.3(+/-0.7)x10(4) cfu/ml, respectively. Microbial attachment in the non-pressurised tank of the hydraulic system increased with increasing pipeline pressure [from 1.0(+/-0.3) to 3.8(+/-2.7)x10(5) cells/cm(2) on stainless steel]. The phosphatase, aminopeptidase and beta-glucosidase activities in the pressurised medium were between 0.02 and 1.4 micromol/lh ( V(max)) and decreased in response to increasing pipeline pressure. The alpha-glucosidase activity was detected only at 70 bar and the glucuronidase activity only occasionally. Based on principal component and cluster analyses, both the pressure applied and the original filling water quality affected substrate utilisation patterns. This study demonstrated the capability of freshwater bacteria to tolerate high and fluctuating pressure in a technical water system. Microbial survival was due to attachment and growth on the surfaces of the non-pressurised components and the nutrient flux released by cell lysis in the pressurised components. In summary, high pressures in water hydraulic systems do not prevent potential microbiologically related operational problems.

Bacteria↗

Performance evaluation of a low conductive growth medium (LCGM) for growth of healthy and stressed Listeria monocytogenes and other common bacterial species.

The performance of a low conductive growth medium (LCGM) (conductivity of <1300 microS) was evaluated for its ability to support growth of food borne bacterial pathogens including Listeria monocytogenes and to determine the expression of the two key virulence proteins in L. monocytogenes for possible applications in an impedance-based microfluidic biochip detection platform. Growth of Listeria was monitored spectrophotometrically and the lag phase, generation time, growth rate and maximum population density were determined using the Gompertz equation. LCGM had a lag phase of 2.3 h and showed a higher cell density compared to Luria Bertini (LB) broth. Length of lag phase was highly dependent on initial inoculum concentrations. The changes in conductivity with respect to growth in the low conductive medium were monitored using a conductivity probe. L. monocytogenes growth could be detected within 2 h (0.1 mS) in LCGM and within 6 h in LB. The performance of the media was also evaluated for the recovery of Listeria cells exposed to various stresses as 42 degrees C for 1, 2 or 6 h, an osmotic stress in 10.5% NaCl, an acidic stress at pH 2, 3 or 5 and a combined stress of 10.5% NaCl, pH 5 and 1 h exposure at 42 degrees C. The recovery rate was comparable with that of Tryptic soy broth containing yeast extract (TSBYE). L. monocytogenes in LCGM supported the expression of two key virulence markers, actin polymerization protein (ActA) and internalin B (InlB), which could be detected using specific antibodies. In general LCGM also supported the growth of several other bacterial species suggesting its implication in microbial quality monitoring of products. In conclusion, LCGM is a sensitive low conductive medium that supports the growth as well as the expression of virulence markers for potential applications in sensitive detection of L. monocytogenes or other food borne pathogens in impedance-based sensor platform.

Colony Count, Microbial↗

Microbiologic spectrum and susceptibility of isolates: part I. Postoperative endophthalmitis. Endophthalmitis Research Group.

PURPOSE: To present the microbial spectrum and susceptibilities of isolates in postoperative endophthalmitis. METHOD: Isolates from 206 eyes of 206 patients who underwent vitrectomy for postoperative endophthalmitis were examined. RESULTS: One-hundred twelve (54.4%) of 206 vitreous samples were culture positive and 14 (12.5%) of 112 culture-positive cases were polymicrobial, yielding a total of 126 isolates. Isolates included 59 (46.8%) gram-positive cocci, eight (6.3%) gram-positive bacilli, 33 (26.2%) gram-negative organisms, five (4.0%) Actino-mycetes-related organisms, and 21 (16.7%) fungi. Susceptibilities to amikacin, ceftazidime, chloramphenicol, cefazolin, ciprofloxacin, gentamicin, and vancomycin are reported. CONCLUSIONS: This is the largest, single-center, prospective series on microbial susceptibilities in postoperative endophthalmitis. We report a high prevalence of gram-negative species and fungi, suggesting that empiric therapy should include coverage for gram-negative pathogens and for fungal pathogens in appropriate settings.

Anti-Bacterial Agents↗

A comparison of enumeration methods for culturable Pseudomonas fluorescens cells marked with green fluorescent protein.

The detection of bacteria in environmental samples using genetic markers is valuable in microbial ecology. The green fluorescent protein (GFP) reporter gene was studied under nutrient starvation conditions at 4 degrees C, 23 degrees C and 30 degrees C in Pseudomonas fluorescens R2fG1 cells tagged with a red-shifted gfp. Fluorescence intensity was not significantly different in cells maintained in a buffer for at least 48 days at all the tested temperatures. gfp-Tagged R2fG1 cells were introduced into bulk soil microcosms and soil microcosms with wheat seedlings. GFP-marked cells were enumerated immediately after inoculation into soil and again in soil and root samples after 10 days. Counts of culturable colonies were obtained from drop plates using 5-microl aliquots of serial dilutions viewed with an epifluorescent microscope. Traditional spread plates (using 100-microl aliquots) and the most-probable-number (MPN) method using a spectrofluorometer were also used to enumerate the GFP-marked Pseudomonas cells in soil, rhizosphere and rhizoplane samples. Microcolonies were visualized on root surfaces under the epifluorescent microscope after immobilizing in agar and incubation for 24 h. Counts from traditional spread plates were significantly higher (P<0.05) than the population estimates of the MPN method for all treatments at any sampling time. Counts using the drop plate method, however, were not significantly different (P<0.05) except in one treatment, and provided similar estimates in half the time of spread plates and at an estimated third of the cost.

Bacteriological Techniques↗

Reduction of the native microflora on alfalfa sprouts during propagation by addition of antimicrobial compounds to the irrigation water.

Alfalfa and other types of sprouts are known to harbor large populations of native microorganisms. As some of these microbes may be causes of reduced shelf life of the product (plant pathogens and other spoilage organisms) and sprouts may, on occasion, harbor bacteria pathogenic towards humans, the addition of antimicrobial compounds to the irrigation water may be warranted. In this study, we tested the efficacy of several antimicrobial compounds for reducing the native microbial populations on alfalfa sprouts during propagation. These compounds included H2O2, peroxyacetic acid+hydrogen peroxide (Tsunami 100), acidified NaClO2, NaClO2 (Aquatize), EDTA, Na3PO4 and NaOCl. When added to the irrigation water at vanous concentrations, none of the antimicrobial compounds reduced the levels of any class of native microflora by more than 1 log10 without evidence of phytotoxicity.

Colony Count, Microbial↗

Generation of larger numbers of separated microbial populations by cultivation in segmented-flow microdevices.

The high speed production of fluid segments for the highly parallelized cultivation of monoclonal cell populations was carried out by the use of microchip segmentor modules. Aqueous fluid segments, embedded in a non-miscible carrier liquid, were produced with frequencies up to 30 s(-1) and showed a high homogeneity in size. This corresponds with the production of about 2.5 million samples per day. The segment volumes can be adapted between about 4 nl and 100 nl. The typical segment size for cultivation experiments is in the range between 40 nl and 80 nl. Nutrient medium can be applied instead of pure water. It is possible to aliquot a cell suspension in such a way that most of the aqueous fluid segments contain only one cell. In model experiments with four microbial species chip-produced aliquots of 60 nl, each containing one or a few cells, were incubated in Teflon capillary tubes. Rapid growth of the microcultures was observed. Cell densities were found to be as high as in conventional shake flask cultures.

Colony Count, Microbial↗

Foreign body associated infection.

One of the major complications associated with the use of medical devices and implants is foreign body associated infection. Its significance in modern medicine and its pathogenesis are discussed. Adherence to and colonization of a foreign body material and the interference of microbial products with the host defence are critical steps leading to foreign body infections. Therapy is difficult and hence the immediate administration of antibiotics and the removal of an infected device or implant is still preferred. New strategies for the prevention of foreign body infections are briefly introduced.

Anti-Bacterial Agents↗

Extension of shelf life of whole and peeled shrimp with organic acid salts and bifidobacteria.

Microbiological and sensory characteristics of treated whole and peeled shrimp from the east coast of Saudi Arabia were evaluated. Shrimp samples were treated with organic acid salts with or without Bifidobacterium breve culture and stored in ice. Peeling alone extended the microbiological shelf life by 4 days. Treatment of whole shrimp with sodium acetate alone or potassium sorbate with bifidobacteria prolonged the microbiological shelf life by 3 days and increased the microbial generation time from 12.8 h (control) to 30.1 h or 31.4 h, respectively. The microbiological and sensory shelf life of peeled shrimp treated with sodium acetate was more than 17 days. Sodium acetate extended the microbial lag phase and lengthened the generation time (38.7 h compared to 15.8 h for the control). Micrococci and coryneforms were the predominant microorganisms in whole shrimp during storage. Treatment with sodium acetate maintained better sensory characteristics for peeled shrimp than potassium sorbate combined with bifidobacteria.

Animals↗

Effect of microorganism characteristics on leak size critical to predicting package sterility.

The effects of microorganism size and motility on the leak size critical to the sterility of a package, along with the imposed pressure required to initiate liquid flow for the critical leak size, were measured. Pseudomonas fragi Lacy-1052, Bacillus atrophaeus ATCC 49337, and Enterobacter aerogenes ATCC 29007 were employed to assess package sterility. One hundred twenty-six 7-mm-long microtubes with interior diameters of 5, 10, and 20 microm were used to simulate package defects. Forty-two solid microtubes were used as controls. No significant differences were found between sizes or motility statuses of test organisms with respect to loss of sterility as a result of microbial ingress into test cells with microtube interior diameters of 5, 10, and 20 microm (P > 0.05). Interactions between the initiation of liquid flow as a result of applied threshold pressures and sterility loss for test cells were significant (P < 0.05).

Bacillus↗

Thiobacillus sp. W5, the dominant autotroph oxidizing sulfide to sulfur in a reactor for aerobic treatment of sulfidic wastes.

The floating filter technique was successfully adapted for the isolation of the dominant, chemolithoautotrophic, sulfide-oxidizing bacterium from a sulfur-producing reactor after conventional isolation techniques had failed. The inoculated polycarbonate filters, floating on mineral medium, were incubated under gaseous hydrogen sulfide at non-toxic levels. This technique gave 200-fold higher recoveries than conventional isolation techniques. Viable counts on the filters, making up 15% of the total count, appeared to be all of the same species. Chemostat cultures of the new isolate had a very high sulfur-forming capacity, converting almost all hydrogen sulfide in the medium to elemental sulfur under high sulfide loads (27.5 mmol l-1 h-1) and fully aerobic conditions. This behaviour closely resembled that of the microbial community in the sulfur-producing reactor. Moreover, similar protein patterns were obtained by electrophoresis of cell-free extracts from the isolate and the mixed culture. It has therefore been concluded that this isolate represents the dominant sulfide-oxidizing population in the reactor. The isolate has been shown to be a new Thiobacillus species, related to Thiobacillus neapolitanus. In view of the general confusion currently surrounding the taxonomy of the thiobacilli, a new species has not been formally created. Instead, the isolate has been given the working name Thiobacillus sp. W5.

Aerobiosis↗

Color measurement as a means of quantifying surface biofouling.

Laboratory reactors fitted with removable ceramic porcelain growth surfaces were inoculated with a consortium of biofilm forming environmental isolates. A Minolta colorimeter CR-200 (Minolta Camera Co., Ltd, Ramsey, NJ) was used in conjunction with a specially designed adapter to evaluate the reflective color of the porcelain disks as biofilm accumulated on them. Areal viable cell counts were monitored over a period of eleven days in two separate experiments and direct color measurements of the untreated, microbially fouled test surfaces were collected. This colorimetric assay was both non-destructive and immediate. A strong linear relationship between log cell density and log color change was observed. The Pearson product moment correlation coefficient for all 45 observations combined was r = 0.95. Separate regression lines for each experiment were not significantly different (P = 0.19). When adjusted for time, the (partial) correlation coefficient between log cell density and log color change was r = 0.87, which suggests that the relationship between the two measures can not be explained by their mutual dependence on time. Reflective color measurement provided a rapid, non-destructive and quantitative measure of biofllm accumulation.

Biofilms↗

Changes in symbiotic and associative interrelations in a higher plant-bacterial system during space flight.

The miniature cenosis consisting of the water fern Azolla with its associated symbiotic nitrogen-fixing cyanobacterium Anabaena and the concomitant bacteria was investigated. Ecological closure was shown to produce sharp quantitative and qualitative changes in the number and type of concomitant bacteria. Changes in the distribution of bacterial types grown on beef-extract broth after space flight were recorded. Anabaena azollae underwent the most significant changes under spaceflight conditions. Its cell number per Azolla biomass unit increased substantially. Thus closure of cenosis resulted in a weakening of control over microbial development by Azolla. This tendency was augmented by spaceflight factors. Reduction in control exerted by macro-organisms over development of associated micro-organisms must be taken into account in constructing closed ecological systems in the state of weightlessness.

Anabaena↗

A laboratory evaluation of the decontamination properties of microfibre cloths.

Standards of cleanliness in health care continue to attract attention. Effective cleaning requires the input of energy, and microfibre cloths may help in the physical removal of soil. The ability of these cloths to remove organic soil (measured by ATP) and bacteria was compared with paper towel and a conventional cloth in controlled wet and dry conditions. When used wet on a dry surface, the cleaning ability of six different microfibre cloths was variable, and in most cases, not significantly better than paper towel or a conventional cloth. One type of microfibre cloth did perform significantly better than the others and paper towel in reducing both organic soil and microbial load. When used dry on a dry surface, there was no significant difference between the cloths, and none of the cloths reduced microbial and organic bioburden effectively. The ability of the cloths to recontaminate the surface was also tested, and some of the microfibre cloths transferred significantly less organic debris and micro-organisms back to the surface than other cloths. Different makes of microfibre cloths have different characteristics, and the name 'microfibre' should not imply superior cleaning efficacy.

Colony Count, Microbial↗

Comparison of media and a liquid detergent extraction step on bacterial recovery from animal feeds.

The objective of this study was to evaluate the recovery of Tryptic soy agar (TSA) or low nutrient R2 bacterial populations from fresh feeds and animal feeds held in long-term cold storage after extraction with a detergent-based solution combined with repeated centrifugation. Repeated centrifugation after exposure to a detergent-based solution resulted in recoveries generally no more than 10% of the microbial population in the original feed. Feed R2A populations were significantly correlated (P<0.01) with TSA populations enumerated from the feed and feed extracts, as well as R2A populations from feed extracts. Similarly, TSA bacterial populations from feed were significantly correlated with TSA populations from extracts.

Animal Feed↗

A simple approach to examine early oral microbial biofilm formation and the effects of treatments.

BACKGROUND/AIMS: A simple in vivo approach to examine early dental plaque formation in the human mouth and to determine the effects of common dietary and oral hygiene procedures on biofilm formation is reported. METHODS: A custom designed device that fits securely behind the teeth of the mandibular arch provides a surface for microbial colonization. This device is prepared with denture acrylic and can be repeatedly used by the subject, exposing a large and constant surface area for microbial accumulation. RESULTS: Large numbers of oral bacteria colonized the device by 2 h; these increased significantly by 4 h (P < 0.05). Bacterial colonization increased significantly after rinsing with a sucrose solution (P < 0.05) but remained unaffected after rinsing with water, a commercially available fluoride mouthrinse without antimicrobial agents, or brushing with a fluoride dentifrice (P > 0.05). Rinsing with mouthrinses formulated with chlorhexidine, cetylpyridinium chloride or triclosan/copolymer significantly inhibited colonization (P < 0.05). A dose-dependent inhibition was noted with chlorhexidine rinses (P < 0.05). Brushing with a triclosan/copolymer dentifrice significantly inhibited microbial colonization compared with a control (P < 0.05). CONCLUSION: This simple approach was useful for examining the effects of common dietary and oral hygiene procedures. Significant biofilm inhibitory effects were noted with formulations that demonstrated efficacy in previous clinical studies.

Actinomyces viscosus↗

Treponema denticola and Porphyromonas gingivalis as prognostic markers following periodontal treatment.

Subgingival plaque samples were collected from individuals with advanced periodontitis before and 3 to 11 weeks after scaling and root planing periodontal treatment. The plaque levels of Treponema denticola and Porphyromonas gingivalis antigens were measured before and after treatment by a quantitative immunoassay procedure using monoclonal antibodies specific for these oral bacteria. A decrease in mean levels of T. denticola (P less than .05) and P. gingivalis antigens (P less than .09) were observed following periodontal therapy. Improved health, as measured by a decrease in probing depth, was associated with a decrease in T. denticola antigen (P less than .05). These results suggest that the T. denticola levels of successfully treated sites decreased, while non-responding sites had levels of this microbial marker which were equal to or greater than the pre-treatment levels. These results provide additional evidence that T. denticola is associated with human adult severe periodontal disease, and can serve as a prognostic marker for disease recurrence.

Adult↗