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Microorganisms develop biofilms on indwelling medical devices and are associated with device-related infections, resulting in substantial morbidity and mortality. This study investigated the effect of pretreating hydrogel-coated catheters with Pseudomonas aeruginosa bacteriophages on biofilm formation by P. aeruginosa in an in vitro model. Hydrogel-coated catheters were exposed to a 10 log10 PFU ml−1 lysate of P. aeruginosa phage M4 for 2 h at 37°C prior to bacterial inoculation. The mean viable biofilm count on untreated catheters was 6.87 log10 CFU cm−2 after 24 h. The pretreatment of catheters with phage reduced this value to 4.03 log10 CFU cm−2 (P < 0.001). Phage treatment immediately following bacterial inoculation also reduced biofilm viable counts (4.37 log10 CFU cm−2 reduction; P < 0.001). The regrowth of biofilms on phage-treated catheters occurred between 24 and 48 h, but supplemental treatment with phage at 24 h significantly reduced biofilm regrowth (P < 0.001). Biofilm isolates resistant to phage M4 were recovered from catheters pretreated with phage. The phage susceptibility profiles of these isolates were used to guide the development of a five-phage cocktail from a larger library of P. aeruginosa phages. The pretreatment of catheters with this cocktail reduced the 48-h mean biofilm cell density by 99.9% (from 7.13 to 4.13 log10 CFU cm−2; P < 0.001), but fewer biofilm isolates were resistant to these phages. These results suggest the potential of applying phages, especially phage cocktails, to the surfaces of indwelling medical devices for mitigating biofilm formation by clinically relevant bacteria.
Indwelling medical devices of various kinds may become colonized with microorganisms, resulting in the formation of microbial biofilms (16). Biofilm-associated organisms are tolerant to antimicrobial agents, can evade the host immune system, and can act as a nidus for infection (16). As a result, device-related infections, such as catheter-associated bloodstream infections, cause substantial morbidity and mortality among specific patient populations (9). Attributable mortality rates for healthcare-associated bloodstream infections have been estimated to be 25% (44).
A number of novel strategies have been proposed to more effectively prevent and control device-associated biofilms, either by minimizing microbial attachment to device surfaces or by targeting the biofilm after it has developed. One such strategy is to use bacteriophages (phages) (17). Phages have been used for the treatment of infectious diseases in plants (26), animals (6), and humans (33, 39, 43). The use of phages to control biofilms has potential for several reasons. Phages can replicate at the site of an infection, thereby increasing in numbers where they are most required. During the lytic replication cycle, the infection of a bacterial host cell by a single phage virion will result in the production of dozens or hundreds of progeny phage, depending on the particular phage and host strains. Some phages also have been shown to produce enzymes that degrade the extracellular polymeric substance (EPS) matrix of a biofilm (23, 25). Doolittle et al. (19) showed that progeny phage will propagate radially through a biofilm. At least in theory, a single phage dose should be capable of treating a biofilm infection as progeny phage infect adjacent cells and degrade the biofilm matrix.
Curtin and Donlan (13) demonstrated that a phage that is active against Staphylococcus epidermidis could be incorporated into a hydrogel coating on a catheter and significantly reduce biofilm formation by this organism in an in vitro model system. Based on those studies with S. epidermidis, we have investigated whether phages specific for Pseudomonas aeruginosa also can reduce biofilm formation by this organism in a similar in vitro model. Catheters were treated with a single phage or a combination of phages prior to, immediately following, or 24 h after inoculation with the test P. aeruginosa culture, and the effects of the phage treatments on biofilm formation and maintenance were characterized.
(Portions of this paper were presented as poster no. A-011 at the 2007 American Society for Microbiology General Meeting in Toronto, Canada [20a].)
Organisms and culture conditions. P. aeruginosa M4 (Health Protection Agency, Colindale, United Kingdom) was selected as the biofilm-forming organism in this study. Stocks were maintained at −80°C and propagated on Trypticase soy agar (TSA) plates at 37°C unless otherwise indicated.
P. aeruginosa phage M4 (Health Protection Agency, Colindale, United Kingdom) was used for the first round of phage treatment experiments. Phage M4 (35) is a member of the family Myoviridae (1). By using a single-step growth curve (21), this phage was shown to exhibit a latent period (the time from initial infection to lysis) of approximately 35 min and a burst size (number of progeny phage per infected cell) of 95 to 100. Phage M4 stocks were maintained as lyophilized preparations that were stored at 4°C.
Environmental phages were incorporated into the phage cocktail that was used for the second round of biofilm treatment experiments. These phages were isolated from the Snapfinger Creek Wastewater Treatment Plant in Dekalb County, GA. Untreated sewage samples were allowed to settle overnight at 4°C and centrifuged at 9,000 × g for 20 min, after which the supernatant was filter sterilized (0.22-μm pore size; Millipore, Billerica, MA). Twenty milliliters of 2× tryptic soy broth (TSB), 20 ml of the filtered sewage, and 0.4 ml of a log-phase TSB broth culture of one of seven clinical or laboratory strains of P. aeruginosa were combined and incubated at 37°C overnight at 100 rpm in a shaker incubator. Samples that were cleared (indicative of the phage lysis of the bacterial culture) were centrifuged at 4,000 × g for 30 min, filter sterilized (0.22 μm), and stored at 4°C. The titer of the crude phage lysate was determined by plaque assay using the soft-agar overlay method on TSA (2). Ten phage isolates were recovered and used for host susceptibility testing and phage cocktail preparation.
Three of the Colindale Pseudomonas typing phages, M6, F8, and Col11 (35) (Health Protection Agency, Colindale, United Kingdom), also were used for susceptibility testing.
Phages were propagated using the soft-agar overlay method (2). Larger volumes of high-titer phage lysates were prepared as described by Adams (2) using P. aeruginosa M4 as the host strain in TSB (Difco, Becton Dickinson, Sparks, MD) amended with 3 mM MgCl2 (added as MgCl2·6H2O) and 4 mM CaCl2 (added as CaCl2·2H2O). Briefly, 1 ml of an overnight culture of P. aeruginosa M4 (grown at 37°C) was added to 49 ml of TSB containing supplemental MgCl2 and CaCl2, and the culture was incubated at 37°C with shaking at 250 rpm. When the optical density at 550 nm was approximately 0.3 (approximately 109 CFU/ml) using a Microscan turbidity meter (Dade Behring, West Sacramento, CA), M4 phage was added to a final concentration of 109 PFU ml−1. The culture was allowed to stand for 15 min at room temperature and then incubated for 18 h at 37°C with shaking at 100 rpm. Host cell debris was pelleted by centrifugation (4,100 × g for 35 min), and the supernatant containing phage was filter sterilized (0.22-μm pore size; Millipore, Billerica, MA) and stored at 4°C. The titer of the crude phage lysate was determined using the soft-agar overlay method on TSA (2).
In vitro model system for growing biofilms on hydrogel-coated catheters.Biofilms were grown on Lubri-sil all-silicone 16 French Foley catheters (C. R. Bard, Covington, GA) in a modified drip flow reactor (mDFR) (Biosurface Technologies, Bozeman, MT). These catheters contained a neutral hydrogel coating on external and luminal surfaces. The mDFR has four chambers, each with a sealing lid. The original device was modified to allow the connection of catheter segments to the influent and effluent ports within the device. Following sterilization by ethylene oxide gas, the catheters in the mDFR each were connected by means of silicone tubing to glass vessels containing (i) phage lysate, (ii) bacterial inoculum, (iii) sterile medium, and (iv) an empty waste container. The entire system was assembled and operated in a 35°C incubator.
For experiments examining biofilm formation on non-phage-treated catheters, an overnight culture of P. aeruginosa M4 in 100% TSB was pumped through the catheters at 1 ml min−1 for 2 h using a Masterflex pump (Cole Parmer, Niles, IL) to allow the organisms to attach to the catheter surface. This was followed immediately by pumping sterile 10% (vol/vol) TSB through the catheters at 0.5 ml min−1 for an additional 22 or 46 h depending upon the experimental design used. The mean concentration of the bacterial inoculum ranged from 108 to 109 CFU ml−1 during the 2-h inoculation period.
Phage treatment of the catheter surfaces.Three types of phage treatments were used: pretreatment, posttreatment, and recharge treatment. Phage pretreatment and posttreatment involved exposing the catheter lumen to a phage lysate for a defined period of time. In both cases, a crude phage lysate (prepared as described above) with a titer of 1.0 × 1010 to 2.2 × 1010 PFU ml−1 was used. For pretreatment experiments, the phage lysate was pumped through the catheter segments for 2 h at 1 ml min−1 prior to exposure to the bacterial inoculum for 2 h, followed by sterile medium for either 22 or 46 h. For posttreatment experiments, catheters were exposed first to bacterial inoculum for 2 h, phage for 2 h, and then sterile medium for 22 or 46 h. For recharge experiments, catheters were pretreated with phage for 2 h, exposed to bacterial inoculum for 2 h, exposed to sterile medium for 22 h, and treated again with phage by pumping the high-titer phage lysate through the catheters for 2 h and then sterile medium for an additional 22 h.
Two rounds of phage treatment experiments were conducted. In the first round, pretreatment, posttreatment, and recharge experiments were conducted using phage M4. In the second round, pretreatment experiments were conducted using a phage cocktail that consisted of equal numbers of each of M4 phage and four environmental phages (ΦE2005-24-39, ΦE2005-40-16, ΦW2005-24-39, and ΦW2005-37-18-03) for a final cocktail titer of 7.0 × 109 PFU ml−1. The phages in this cocktail were selected based on the phage susceptibility profiles of biofilm isolates recovered from the first round of phage treatment experiments.
Two sets of control experiments also were conducted. Catheters were exposed for 2 h to heat-inactivated M4 phage (80°C for 3 h) as a control pretreatment. Also, a conditioning serum film was simulated on the catheter lumen to assess its effect on biofilm formation and phage M4 efficacy. Filter-sterilized whole human serum (complement inactivated at 56°C for 30 min) was instilled into the lumen of phage-pretreated catheter segments in the mDFR and incubated for 2 h at 35°C (13). The experiment continued with the 2-h bacterial inoculation and 22-h sterile medium incubation periods. The catheter segments were sampled and processed to assess biofilm development, and results were compared to results for untreated catheters not exposed to serum.
Recovery and enumeration of biofilm organisms.Catheters were removed from individual chambers of the mDFR at different time points following bacterial inoculation. In the case of untreated and pretreated catheters, catheter samples were collected after 2, 6, 24, and, in some experiments, 48 h after bacterial inoculation. For posttreatment catheters, samples were collected 4, 6, 24, and 48 h after bacterial inoculation. It was not possible to collect 2-h samples from these systems, since phage posttreatment immediately followed bacterial inoculation.
The ends of the catheter segments were clamped shut with hemostats and cut from the mDFR fittings. All processing was conducted aseptically. Fluid from the catheter lumen was collected to quantify phage in the luminal fluid by the soft-agar overlay method. The catheter was cut into two 1-cm sections. Each section then was cut lengthwise, rinsed gently in phosphate-buffered saline (PBS), and placed into a 50-ml sterile centrifuge tube containing 10 ml PBS. Biofilm organisms were recovered from catheter segments using the following method: tubes containing catheter sections were sonicated at 42 kHz in a water bath sonicator (Branson 2510; Branson, Danbury, CT) for 10 min, vortexed for 30 s, sonicated again for 5 min, vortexed for 30 s, sonicated a third time for 30 s, and then vortexed a final time for 30 s. This recovery method was similar to the method described by Curtin and Donlan (13), which was shown to recover essentially all viable biofilm cells from the catheter surface. The recovered biofilm cells then were quantified by serial dilution spread plating on TSA, and viable counts were determined and expressed as CFU cm−2 of the catheter lumen surface. Organisms recovered from the biofilms were subcultured on TSA and confirmed as P. aeruginosa by the Vitek Legacy GNI+ card (bioMerieux, Durham, NC).
To determine the concentration of phage present on the catheter lumen surface prior to exposure to the bacterial inoculum, a 1.2 × 1010 PFU ml−1 phage M4 lysate in cation-supplemented TSB was pumped at 1 ml min−1 through each of four catheter sections contained in the mDFR for 2 h at 35°C. Catheter sections were clamped and removed from the reactor and processed according to the protocol used to recover biofilm cells, and the phage concentration was determined by plaque assay.
Effect of catheter processing procedure on phage viability and assessment of phage effect on P. aeruginosa during sampling procedure.The effect of the processing method itself on phage viability was determined by adding 1 ml of a 5 × 1010 PFU ml−1 M4 phage lysate to each of six 9.0-ml Butterfield buffer tubes, exposing three of these tubes to the sonication and vortexing process described above, and enumerating PFU in all six tubes using the soft-agar overlay method (2).
To demonstrate that phage M4 was not causing the further lysis of P. aeruginosa M4 during the biofilm-sampling procedure, a suspension equivalent to a 0.5 McFarland standard (1 × 108 CFU ml−1) of P. aeruginosa M4 was prepared in 5 ml Butterfield buffer at 4°C. A 1-ml aliquot was removed for viable cell count determination, and 1 ml of 1010 PFU ml−1 phage M4 lysate was added to the remaining 4-ml solution. The tube was subjected to the same biofilm removal procedure as that used during mDFR experiments (i.e., sonication and vortexing), followed by chilling on ice for 2 h (which was twice the length of time required for sampling during the biofilm/phage experiments). The number of viable cells then was determined by plate count. Counts of untreated and phage-treated cell suspensions were compared. This experiment was performed in triplicate.
Characterization of phage-resistant biofilm isolates.Several colonies of biofilm organisms, representing different colony morphologies recovered from M4 phage-treated, cocktail-treated, and untreated catheters, were collected after 2, 24, and 48 h, subcultured, and characterized to assess resistance to phage M4. All isolates were gram-negative rods and were oxidase positive, and they produced a grape-like odor, pyoverdin, and a yellow-green pigment on Pseudomonas Agar F (Difco Laboratories, Becton Dickinson Co., Sparks, MD), which is indicative of P. aeruginosa (7). Isolates exhibiting atypical reactions were reconfirmed using the Vitek GNI+ card. The susceptibility of each isolate from the first round of phage treatment experiments to phage M4 was determined by the spot test (8) and the soft-agar overlay method (2). To determine whether phage resistance was related to a stress response of the organism or a stable characteristic, isolates were subcultured several times during 7 days in TSB and then tested again for susceptibility to phage M4. The biofilm isolates recovered from phage cocktail-treated catheters in the second round of experiments were tested for susceptibility to each of the five phages in that cocktail using the spot test.
Phage-resistant isolates from both rounds of experiments were tested for their ability to adsorb M4 phage using a modification of the methods of Hadas et al. (22) and Karlson (27). Briefly, an overnight culture of each isolate was grown in TSB to an optical density at 550 nm of 0.12 (approximately 3 × 108 CFU ml−1), and 1 ml of this culture was combined with 0.1 ml of phage M4 (concentration, 3 × 108 PFU ml−1) in a glass tube so that the proportion of phage to bacteria was 1:10 (multiplicity of infection [MOI] = 0.1). This tube was held in a 30°C water bath for 4 min to provide time for phage to adsorb to the bacterial cells. An aliquot then was immediately transferred to a tube of TSB containing five drops of chloroform, which would have no effect on unattached phage particles but would kill bacterial cells and prevent plaque formation by any phage particles adsorbed to those cells. Unadsorbed phage were quantified by the soft-agar overlay method, and the number of adsorbed phage was calculated as the difference between the initial phage count prior to the addition of bacteria and the phage count after the 4-min adsorption period.
Additional characterization tests were conducted on the biofilm isolates recovered from catheters treated with the phage cocktail. Certain isolates were exposed to 50 μg/ml mitomycin C (Roche Applied Science, Indianapolis, IN) to test for the presence of temperate phages (42). Isolates also were tested for the presence of integrated Pf4 filamentous phages using PCR (32).
Relative growth rates for certain biofilm isolates were measured by inoculating TSB with a log-phase culture, incubating it at 35°C at 100 rpm, and measuring absorbance at 600 nm hourly for 6 h using a Hach DR 4000 spectrophotometer (Hach Co., Loveland, CO).
Biofilm formation was assessed using a modified crystal violet assay (11, 34). Briefly, microtiter plates were inoculated with an 18-h culture grown in 10% TSB, incubated at 35°C for 10 h, and stained with an aqueous solution of 1% (vol/vol) crystal violet and 3% (vol/vol) glacial acetic acid. After 15 min, plates were washed several times with sterile water to remove excess stain. Biofilm formation was quantified by the addition of 95% ethanol to each well (200 μl each). The A540 of each tube containing the extracted crystal violet was measured using a Hach DR 4000 spectrophotometer.
The stability of biofilm variant colony morphology was determined by subculturing colonies in brain heart infusion broth through 10 serial transfers according to the method of Häussler et al. (24). Reversion to the M4 parent colony morphology was assessed by growth characteristics on Columbia blood agar.
SEM.Biofilm development on catheter lumen surfaces was visualized by scanning electron microscopy (SEM). Catheter segments were rinsed gently in sterile PBS and then fixed in 5% (vol/vol) glutaraldehyde in 0.67 M cacodylate buffer (pH 6.2) for 1 h and dehydrated through a graded series of 10-min ethanol immersions (30 to 100%). Specimens were immersed overnight in hexamethyldisilazane (Polysciences Inc., Warrington, PA), mounted on aluminum stubs with silver paint (Ted Pella Inc., Redding, CA), coated with gold (Polaron SC7640 sputter coater; Thermo VG Scientific, United Kingdom), and photographed with an ESEM (Philips XL-30; FEI Co., Hillsboro, OR). The entire luminal surface of 1-cm−2 catheter segments was examined, and images were chosen that represented the typical field of view.
Statistical analysis.All experiments were performed a minimum of three times. Bacterial and phage counts were log10 transformed, and differences in microbial recovery were analyzed using two-tailed t tests (Sigma Stat 3.5; Systat Software Inc., San Jose, CA). P values of <0.05 were considered significant.
Effect of P. aeruginosa M4 phage treatment on biofilm formation on catheter surfaces.Pretreatment of hydrogel catheters with 10.07 log10 PFU ml−1P. aeruginosa phage M4 for 2 h at 1 ml min−1 resulted in a mean concentration (n = 4) of 6.16 log10 PFU cm−2 associated with the catheter luminal surface. The method used to recover and quantify phages on the catheter surface was shown to have no significant effect on phage viability when treated and untreated phage lysates were analyzed (P = 0.650). P. aeruginosa M4 attached to untreated hydrogel-coated catheter surfaces and formed biofilms, attaining mean cell densities of 6.87 log10 CFU cm−2 after 24 h of exposure in the catheter model system (Fig. 1). The pretreatment of catheters with phage M4 significantly reduced biofilm formation by P. aeruginosa M4 to 4.03 log10 CFU cm−2 (P < 0.001) after 24 h, a >99.9% reduction. The posttreatment of catheters with phage (i.e., phage treatment immediately following exposure to the bacterial inoculum) also significantly reduced biofilm formation (4.37 log10 CFU cm−2 [P < 0.001]), although to a lesser extent than pretreatment. When catheters were exposed in the mDFR for an additional 24 h (see Fig. 3), biofilm levels on treated catheters increased so that the treatment resulted in overall reductions of less than 1 log10 CFU cm−2 at 48 h, even though this difference between treated and untreated catheters still was statistically significant (P = 0.01 and <0.001 for pretreated and posttreated catheters, respectively). When pretreated catheters were treated a second time with phage (recharged) at 24 h, biofilm cell densities increased but still were significantly (P < 0.001) lower than counts on untreated catheters (5.58 versus 7.13 mean log10 CFU cm−2; 97% reduction) or pretreated catheters that had not been recharged (5.58 versus 6.44 log10 CFU cm−2) at 48 h. The observation that the recharge treatment of catheters significantly reduced biofilm counts compared to catheters that were not recharged suggested that some, but perhaps not all, of the organisms in the biofilm still were susceptible to phage M4 24 h after inoculation. Phage numbers in the luminal fluid in catheters exposed to P. aeruginosa M4 attained 8.13 log10 PFU ml−1 at 2 h, decreased to 6.95 log10 PFU ml−1 at 6 h, and maintained similar levels through 48 h.
Characterization of biofilm isolates from phage-treated catheters.Eleven biofilm isolates from catheters treated with M4 phage were collected and characterized. Although most exhibited atypical colony morphology, shape, and pigment production, no small-colony variants were observed. All were identified as P. aeruginosa. All except one of the isolates from the 2-, 24-, and 48-h samples were completely resistant to phage M4 upon initial testing. The single exception exhibited only partial resistance (growth of several colonies within the zone of clearing). All isolates lacked the ability to adsorb phage M4, suggesting that these organisms lacked receptors required for phage M4 attachment. All isolates collected from the 2-, 24-, and 48-h time points exhibited complete phage resistance after growth in TSB for 7 days. This suggested that these isolates exhibited stable phenotypes and did not revert to the M4 parent phenotype. Biofilm isolates collected from catheters that were not pretreated with phage also were morphologically diverse, although all were completely susceptible to phage M4.
Effect of serum, heat inactivation of phages, and the sampling procedure on biofilm formation.Subsequent exposure to human serum following phage pretreatment did not significantly affect the ability of the treatment to reduce attachment and biofilm formation by P. aeruginosa M4; the mean log10 CFU cm−2 reduction compared to results for catheters that were not treated with phage or serum was 2.95 (P = 0.012) after 24 h. The pretreatment of catheters with heat-inactivated phage had no observable effect on bacterial attachment and biofilm formation by P. aeruginosa M4 after 24 h (P = 0.497) (data not shown). To ascertain that there was no further phage lysis of P. aeruginosa cells during the sampling procedure, viable bacterial counts were performed on standardized cell suspensions of P. aeruginosa M4, both before and after the suspensions were exposed to phage M4 and treated according to the normal biofilm-processing procedure. No difference in viable cell count was observed (P ≫ 0.05) (data not shown).
SEM.As shown in Fig. 2a, untreated catheters were heavily colonized after 24 h of exposure to P. aeruginosa M4. Exposure to the phage pretreatment reduced biofilm cell densities to the extent that even microcolonies or single cells in representative fields were not visible (Fig. 2b).
Development and testing of phage cocktail.The 11 biofilm isolates described above (designated biofilm variants) that were recovered from the first round of phage M4-treated catheters were tested for their susceptibility to M4, three Colindale typing phages (M6, F8, and Col11), and the 10 environmental phages isolated in this study. The results are shown in Table 1. Phage M4 was effective only against the P. aeruginosa M4 parent strain and 2 of the 11 variants. In contrast, several of the other phages were broadly effective against the phage M4-resistant biofilm variants. Therefore, phages ΦE2005-24-39, ΦE2005-40-16, ΦW2005-24-39, and ΦW2005-37-18-03 were grown to a high titer and combined with phage M4 to formulate a phage cocktail, and this mixture was evaluated for its ability to reduce P. aeruginosa biofilm formation and variant regrowth.
In this second round of experiments, the phage cocktail pretreatment reduced the mean log10 biofilm CFU cm−2 from 7.13 (untreated catheters) to 4.13 (treated catheters) after 48 h (P < 0.001), a 99.9% reduction (Fig. 3). The phage cocktail-treated catheters also contained significantly lower final biofilm counts than catheters that had been recharged at 24 h (4.13 and 5.58 mean log10 CFU cm−2, respectively; P = 0.002). This suggests that a combination of multiple phage strains selected to target resistant bacterial strains within the biofilm can broaden the host range of the phage treatment and more effectively control biofilm formation. However, organisms did colonize phage cocktail-treated catheters and develop biofilms. To determine the basis for this biofilm regrowth in the presence of multiple phage types, representatives of each distinct colony type were collected from biofilm plate counts from each of the three phage cocktail pretreatment studies.
Characterization of biofilm organisms on cocktail-treated catheters.Five distinct colony morphologies were identified, and each was confirmed as P. aeruginosa. Each variant was tested for its susceptibility to each of the five phages in the cocktail mixture. Four of the isolates were completely susceptible to at least one of the five phages in the cocktail, and one (48C) was completely resistant to all five phages. This isolate produced small (∼2 mm), nonpigmented colonies on TSA after 24 h of incubation at 35°C. Isolate 48C was characterized further to determine the stability of its phenotype and the basis for its phage resistance. The phenotype of this isolate was stable; reversion to the original M4 parent morphology did not occur after 10 serial transfers in brain heart infusion broth. The growth rate of 48C (0.083 h−1) was lower than that of the M4 parent strain (0.106 h−1), but biofilm formation for 48C (A540 = 0.063) was significantly higher (P = 0.017) than that for the M4 parent strain (A540 = 0.027). Isolate 48C readily adsorbed phage M4 (94.4% adsorption) in an adsorption assay, suggesting that the mechanism of resistance toward this phage was not due to an alteration of phage receptors by this strain. The exposure of each of the biofilm isolates to mitomycin C did not result in the induction of any phage having lytic activity against the parent P. aeruginosa M4 or the other, non-phage-resistant variants, suggesting that lysogeny was not the mechanism for resistance to phages in the cocktail mixture. All biofilm isolates, including 48C, were negative for Pf4 prophage DNA.
Although there have been several studies published on the effect of phages against biofilms (10, 12, 18, 19, 23, 25, 30, 38), we are unaware of other studies that have used phages to prevent bacterial attachment and initial biofilm formation, rather than attempting to control a mature biofilm. Markoishvili et al. (31) developed a nontoxic polymer film (PhagoBioDerm) that was impregnated with phages, ciprofloxacin, and benzocaine, and it has been used to treat patients with different infectious diseases. Biofilms may be implicated in some of these types of infections; however, an explicit evaluation of PhagoBioDerm against biofilms has not been published.
The results of the present study suggest that the phage pretreatment of catheters can reduce biofilm formation by P. aeruginosa. Treatment with a single phage resulted in a 2.8 log reduction (>99%) after 24 h, which compares favorably to results obtained using catheters that incorporate antimicrobial agents. For example, Raad et al. (36) compared the relative efficacies of several antimicrobial agents in a model system for reducing the colonization of polyurethane central venous catheters by Staphylococcus epidermidis. Reductions in bacterial colonization ranged from 0 to >99% depending on the antibiotic used. Ahearn et al. (3) analyzed adhesion after 2 h by several clinically relevant bacteria onto all-silicone catheters, latex-coated catheters with hydrogel/silver, silicone-coated catheters with hydrogel/silver, and lubricious-coated silicone catheters in an in vitro model. The silver-treated catheters showed the reduced adhesion of all organisms tested, although the percent reductions ranged only from 30 to 95.5%, depending on the organism and type of catheter.
An advantage of phage treatment over treatment with antimicrobial agents is that phages are self replicating in the presence of their host cells and are self limiting in their absence. This ensures that phages are produced continually to infect new bacterial cells as long as the bacteria are able to support their replication, and this is supported by the observation that phage numbers in the luminal fluid were 108 ml−1 at 2 h after addition. The flow of medium following phage pretreatment would be expected to wash out any phage particles that were not associated with the catheter surface. Phage persistence in the catheters for 48 h suggests that progeny phage tend to infect other biofilm cells rather than only planktonic cells. Doolittle et al. (19) observed that the spread of infection by TD4, an Escherichia coli phage, in a flow-cell biofilm resembled plaque formation, in that radial diffusion predominated rather than the downstream flow of progeny phage from lysing cells.
The conditions provided by the in vitro model system in this study differed in several respects from conditions that could be present in vivo. The bacterial inoculum level (108 to 109 CFU/ml) was artificially high to determine whether this treatment could be effective under the most extreme bacterial challenge. A smaller inoculum bacterial cell concentration would be expected to result in a larger effective MOI, which in theory could provide a more significant reduction in bacterial colonization and biofilm formation. Further laboratory studies using different levels of bacterial challenge will be required to define the limits of phage pretreatment technology. It also will be necessary to evaluate this technology in animal model systems and ultimately in human studies to fully assess its clinical relevance. However, the observation that the exposure of phage-treated catheters to human serum did not significantly affect phage efficacy suggests the potential of phage to remain active in the presence of a serum conditioning film under in vivo conditions.
One concern with therapeutic uses of phages is the possibility that organisms in the biofilm will develop resistance to the phages. Our results showed that the strain of P. aeruginosa used in this study contains several subpopulations (variants) with distinct phenotypes, and that phage treatment may select for or induce the expression of those phenotypes that confer resistance to phage M4, allowing biofilms of resistant cells to develop. This was not unexpected, since P. aeruginosa is known to exhibit significant phenotypic variability in biofilms (14, 15, 20). However, we did not observe small-colony variants similar to those reported by Webb et al. (41), nor did we detect the presence of Pf4 prophage genes in variants isolated from phage cocktail-treated catheters. Pf4 was shown to influence the structure and stability of P. aeruginosa PAO1 biofilms (37) and mediate the formation of variants within biofilms.
The resistance of bacteria toward phage may be associated with lysogeny, mutations that affect phage adsorption, restriction modification, or the mechanisms of abortive infection such as the presence of clustered regularly interspaced short palindromic repeats (CRISPRs) in the bacterial genome (4, 5, 17). Lenski and Levin (29) measured bacterial phage resistance rates of 10−8 in a chemostat coinoculated with Escherichia coli and phage T4. Sulakvelidze and Kutter (40) suggested that phage cocktails containing two or more phages could be used to reduce resistance. In the present study, the results obtained when screening the biofilm isolates recovered from phage M4-treated catheters against a collection of phages suggested that it is possible to develop a mixture of phages (phage cocktail) that could increase the effective host range and further reduce biofilm formation. The treatment of catheters with a cocktail of the five best phages from that screening significantly reduced biofilm formation by P. aeruginosa M4 compared to that of treatment with a single phage. However, biofilms on the cocktail-treated catheters also contained several phenotypic subpopulations, and one biofilm isolate (48C) exhibited a stable variant colony morphology and complete resistance to all five phages in the cocktail. This isolate also demonstrated greater biofilm formation potential than the M4 parent strain. The mechanism mediating phage resistance in 48C is unknown.
Through continued screening it might be possible to identify phages with lytic activity against this strain. However, as Lenski has suggested (28), phage have less coevolutionary potential than their bacterial host strains, at least in a closed system, allowing bacterial mutants with no corresponding lytic phage strain to evolve. This suggests that the phage cocktail approach we have described is more effective in applications that provide protection against an initial bacterial challenge rather than applications that require prolonged protection against biofilm formation. However, it is noteworthy that only one of the common phenotypic subpopulations recovered from the phage cocktail experiments was completely resistant to the phages used, whereas all of the variant types recovered from the single-phage treatments were resistant to that phage.
We suggest that the application of phage cocktails to the surfaces of indwelling medical devices, such as intravascular catheters and prosthetic joints, could provide a strategy for the reduction in initial colonization and subsequent biofilm formation by clinically relevant bacteria. However, potential obstacles to the use of this treatment must be carefully considered, including the narrow host range of phage, the resistance of host bacteria to phage, potential for inactivation by the patient's immune system, and the safety of phage preparations in humans (17). Further in vivo studies will be necessary to assess the potential of this approach for the prevention of device-associated infections and to determine whether phage pretreatment can be clinically significant.
Conclusions.The pretreatment of hydrogel-coated silicone catheters with P. aeruginosa phage M4 significantly reduced attachment and biofilm formation by P. aeruginosa M4 in a laboratory model system. The regrowth of biofilms on phage-treated catheters occurred, but recharge treatment and treatment with a mixture of phages having activity against the biofilm organisms significantly reduced the extent of biofilm formation compared to that of untreated catheters or catheters treated once with a single phage. The efficacy of phage pretreatment was not significantly affected by later exposure to human serum, and viable phages continued to be detected in the luminal fluid during the 48-h time course of the experiment. These results suggest the potential of phage application to the surface of indwelling medical devices for the reduction of biofilm formation by clinically relevant bacteria. Further testing using in vivo model systems is needed to evaluate the therapeutic potential of this technology.
We acknowledge Janice Carr for scanning electron micrographs. We also thank Ronny Baxter and Michele Davis of C. R. Bard for providing Foley catheters. We acknowledge Jay Ash and the Snapfinger Creek Water Quality Laboratory staff for assistance in the collection of sewage samples and Tyrone Pitt and Mark Granner (Health Protection Agency, United Kingdom) for phage and bacterial stocks.
This research was supported in part by the appointment of W. Fu and O. Mayer to the Emerging Infectious Diseases Laboratory Fellowship Program administered by the Association of Public Health Laboratories (APHL) and funded by the Centers for Disease Control and Prevention. S. M. Lehman is supported by an American Society for Microbiology/Coordinating Centers for Disease Control and Prevention Postdoctoral Fellowship.
The use of trade names and commercial sources is for identification only and does not imply endorsement by the Public Health Service or the U.S. Department of Health and Human Services.
- Received 15 May 2009.
- Returned for modification 5 September 2009.
- Accepted 2 October 2009.
- Copyright © 2010 American Society for Microbiology


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Join us every Saturday morning for our Shabbat service as we pray together to celebrate this important part of our week. We worship in the chapel and this service is loved by our regulars and occasional visitors. Please note that this is our congregational service and usually does not include the celebration Bar/Bat Mitzvah service (although it Continue Reading »
Daughter of Jonathan and Nanci Lewis
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subset: commoncrawl


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After paying for enough plastic that wore and broke I went the metal way and I love this thing.
Now what I need is a good profile to play with.
Anyone out there have any ideas?
Cobra Driver
ANyone have a good Cougar profile for FSX?
Install help, recommend it, hate it, all about game controlers.
Moderators: rd, mel wilson, gbesoain, Cat1, ricktobin, groundsquirrel, skipperdan
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- Cobra Driver
- Junior SurClaro Member
- Posts: 67
- Joined: Thu Jul 08, 2004 9:45 pm
- Location: Rucker
ANyone have a good Cougar profile for FSX?
Pugnare, Fornicare Au Mort
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I haven't played in a while, but Virgil was a great teammate and medic every time I was on a team with her - easy pickup if you want a talented med who will improve your team environment. Try her out!
|Account Details
|Signed Up
|February 3, 2016
|Last Posted
|October 30, 2019 at 7:34 PM
|689 (0.4 per day)
|Game Settings
|In-game Sensitivity
|Windows Sensitivity
|Raw Input
Refresh Rate
|Hardware Peripherals
|Zowie EC1-A
|CM Quickfire TK (Brown Switches)
|PureTrak Talent Cloth
|Philips SHP9500S
|BenQ XL2411 (main) HP Pavilion 23bw (secondary)
Great teammate and a capable medic! I MISS YOU BUBBZI
friendly gamer :)
Good luck, gamers :)
Now THIS is podracing
I'd like to chime in on a few things:
cini care a lot about these issues but hothouse feels like such a buzzword
It's a buzzword I haven't really seen used before, but it's just another name for an already heavily considered possibility. The problem (in regards to world heating) is not so much pollution like plastic in the oceans, but rather, the release of greenhouse gases into the air. Earth used to be a "hothouse" as described. There was far more carbon dioxide in the air, and as a result, earth's atmosphere trapped more heat. As plants perform photosynthesis, they take carbon dioxide from the air and trap carbon, releasing oxygen. Then they die, and that carbon is trapped underground. After hundreds of millions of years, some is trapped in the form of coal and oil... Which happens to make great fuel. Burning fossil fuels is essentially undoing hundreds of millions of years worth of natural carbon sequestering. Other effects, like polar ice caps melting, reinforce the trend- blue ocean absorbs more light than white ice, thus heating the earth more. Processes like these are what could in theory make the warming irreversible (at least within a fathomable time frame). Without a good solution, the planet can go back to the old "hothouse" it used to be. This process would be (and already has been) harmful to civilization, as well as harmful to existing species of plants and animals.
Climate change won't directly end civilization, but if it continues in its current trend it will cause untold harm, and future generations may look back and wonder why we were so selfish and irresponsible.
variousdiscussion about aggregate impact of individual actions, in comparison to large scale corporate and governmental actions
The big picture is that humans like material things, comfort, and ease of life. This is all well and good, but currently the way we as a society provide that isn't sustainable in the long term. We can through widespread and painful regulation force changes in how we produce those things (ie more renewable energy, less wasteful packaging, etc etc) and we can make sacrifices in our lives (use public transportation more rather than driving, etc etc) to reduce what we use. The current reality isn't conducive to either of those types of change happening enough, especially as more of the world advances and consumes more. We'll probably continue down this path until the whole world really starts "feeling the heat", pun intended. If we're determined, and lucky, enough policy change and lifestyle changes will be made to change this trend- but it doesn't seem likely right now.
I will miss Getawhale hosting :(
tftv is the first step?
Live does not miss.
Thanks for all the good times watching you play, highfive :)
Cool guy! Hunter fit in at an open level easily more than a year ago when I was on a team with him, and he's played a lot since then. He was a good chill teammate. I definitely recommend picking him up as a very flexible sub.
big fragger friendly gamer :D
gg JoshEsports, fun match to end the season
zensanityour team won 5-0 and we're arguing in mumble for some reason
e s e a o p e n
also ggs env
20 minutes on and the mumble has descended into all out war, my return continues to go as planned


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Playing the first stage of the final boss I had a really good rockman and mantis get stuck inside the boss with one hull point left and I couldn't stop them from attacking the room they were in (the separate gun rooms), I was about to pull them out too with this in mind but they were stuck when it was cloaked. I don't know if this is intentional or not but it ruined my chances of finishing the boss with the federation cruiser.
Otherwise im really enjoying the game with over 40 hours sunk into it and only one victory, with the circle ship decked out with uber drones and ion cannons.
Also the new patch mentions new areas and a new ship for the slug people but it doesn't appear as a question mark in the hangar and I haven't encountered any slug people space.
Toggle crew attacking subsystems to avoid being lost
General discussion about the game.
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Illinois politicians want to improve roads and bridges, but can the state afford it?
November 10, 2017
Funding bill passes, but schools are still waiting for answers, dollars
September 8, 2017
State makes first school funding payments
September 7, 2017
Illinois lawmakers congratulate Winston on retirement
April 27, 2017
If the budget crisis is not resolved by July, SIU will be 'running on fumes,' university president says
January 17, 2017
Faculty union signs off on two-year contract with university
December 16, 2016
Future uncertain for southern Illinois homeless agencies
December 13, 2016
How will the loss of longtime higher education legislators affect SIU?
November 15, 2016
Illinois Rep. John Bradley loses long-held seat
November 9, 2016
Paul Schimpf wins open Illinois Senate seat, defeating Sheila Simon
November 8, 2016


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Sorry kids. It's hard trying to be profound everyday. Especially when your brain drooled out your ear when your daughter kicked you in the head to wake you up this morning. SO, here's a survey stolen from Misty (via Jamie via someone else, I'm guessing. Though someone had to have created these originally. So maybe props stop with Jam.).
1. Favorite hobby?
Um. Reading, I s'pose.
2. How do you feel about your ex?
Fine. I don't really feel anything.
3. Do you own guns?
No. But I'm not wholly against them, either.
4. What flavor do you usually drink at Sonic?
Eh. I don't like Sonic much, but I guess the cherry limeade?
5. Do you get nervous before doctor's appts?
Not really.
6. What do you think of hot dogs?
I think they're foul unless I'm at a baseball game or camping.
7. Favorite Christmas song?
O Holy Night
8. What do you prefer to drink in the morning?
A big ole mug of coffee, with lots of milk and sweetness added.
9. Can you do push ups?
I can. Maybe a dozen or so.
10. Can you do a chin up?
Ugh. I doubt it.
12. Do you like blue cheese?
13. Ever been in a car wreck?
Ha. Yes. The real question is how many ambulances have I had called for my wrecks.
14. Do you have ADD?
Only when listening to songs.
15. What's one trait that you hate about yourself?
I'm lazy. And shamefully ignorant in war history – so I discovered during a painful game of Trivial Pursuit last week.
16. Middle name?
17. Name 3 thoughts at this exact moment.
My 16 y/o cousin schooled me in Trivial Pursuit.
I really need to make some coffee.
The air blowing from the vent on my arm is pissing me off.
18. Name 3 things you bought yesterday.
Pop, drinks, chicken
19. Name 3 drinks you regularly drink.
Coffee, water, beer
20. Current worry?
Ahhh. Let me count the ways..
21. Current hate right now?
Uh. The only things I can think of off the top of my head are slow drivers and the fact that I saved my stupid cat's life, and now I just want to kill him.
22. How did you bring in the new year?
Eating sushi and hanging with friends. It was fun.
23. Where would you like to be right now?
Either getting a massage or taking a nap in the mountains. Maybe both.
24. Do you like bok choy?
I don't know.
25. Do you own slippers?
No. My feet get hot easily, so the most I'll wear around the house is socks in the winter.
26. What shirt are you wearing?
A soft t-shirt that says Tree Hugger.
27. Do you like sleeping on satin sheets?
Sure. But not if I have satiny jams on, b/c then they'd get all twisted up and annoying.
28. Can you whistle?
29. Could you be a pirate?
Uh. What?
30. What songs do you sing in the shower?
Whatever's in my head.
31. Favorite girl's name
Lorelei Elizabeth
32. Favorite boy's name
Jacoby Ryan
33. What's in your pocket?
No pockets on yoga pants.
34. Last thing that made you laugh?
A conversation on chat.
35. Best bedsheets as a child?
No idea.
36. Worst injury you ever had?
Knocked my kneecap off to the side, and then had to pop it back on. That hurt like a beyotch. More than childbirth, in fact.
37. Do you like where you live?
38. How many TVs in the house?
39. Who is your loudest friend?
Andy Backhus
40. Favorite book?
To Kill a Mockingbird. I swear.
41. Favorite sports team?
Mmm. Wildcats and the Avs from a few years ago.
42. Song you want played at your funeral?
It changes daily. I have a hundred songs.
43. Favorite candy?
44. Does someone have a crush on you right now?
My cat thinks I am the sun and the moon. Too bad I want to give him away for being annoying.
45. What is the first thing you thought of this morning?
Damn. She just kicked me in the head.


text:
A primary launch is a location with official status reached in agreement with the landholder. This ensures that cartop box access is an officially sanctioned use of that location and that this purpose will be ensured in future plans for this location. It was picked by the BC Marine Trails Network for its strategic location and above-average facilities and convenience.
The primary launch site list is not exhaustive. To see all options, please see the complete launch site list on the main BC Marine Trails Network map.
Remember Me


text:
Loyalty programs are designed for continuous participation from members.
But if you're a loyalty program manager responsible for generating program ROI, it's not enough to simply understand where you are — you need to know where you're going.
Unless you have a crystal ball, however, you're likely going to need some help. But where should you start?
The answer is predictive modeling. Predictive modeling, which uses statistics to predict future outcomes, helps program managers determine the value each loyalty program member will bring to a company. Unfortunately, accurately forecasting the right numbers is easier said than done.
That's where actuaries come in.
While their title may not be the most glamorous, actuaries are the financial experts who can shed light on the future of your loyalty program.
How exactly? By forecasting and extrapolating off past and current data, often with great accuracy. In fact, employing actuarial insights is one of the best ways to prepare your program for long-term financial success.
Actuaries can help you answer important questions such as:
"How can I optimize member acquisition?"
"Who are my highest value members?"
"Is my program strategy driving incremental value?"
When you arm yourself with an actuary, you're taking an essential step towards setting and achieving your loyalty program goals.
In this piece, we'll explain:
- What actuaries actually do
- 9 reasons why actuarial insight is useful to your loyalty program
- Why a data science team isn't enough
- What you should look for in a loyalty program actuary
Do you know where your program is heading and why? Let's enter the world of actuarial insight.
What does an actuary actually do?
To those new to the world of actuarial consulting, actuaries are somewhat of an enigma.
The Society of Actuaries labels the profession as "Part super-hero. Part fortune-teller. Part trusted advisor." While you probably won't encounter an actuary with a cape, these professionals take on an essential role in accounting for loyalty programs: they manage loyalty program risk by planning for the future and protecting their organization against loss.
Actuaries are generally associated with the insurance industry
Most people associate actuaries with insurance, and they're right: over 50% of actuaries are employed directly or indirectly by the insurance industry.
Insurance companies are in the business of predicting outcomes. These companies are required to reserve cash for the policies that they sell, and must set aside money for the claims they expect to eventually pay out on those policies. Given the range of insurance policies, this planning can take on many different forms.
For example, when an insurance company issues your car insurance policy, they have to estimate the probability of paying a claim, the size of that claim, and the timing of when it will occur.
You pay your policy annually. And maybe someday (hopefully not), you'll need to draw on your insurance policy to cover the costs of an accident or damage to your vehicle. The insurance company has to ensure, on average, customers are paying more into the plan than the company is doling out. It's how they stay in business while still providing uninterrupted coverage.
Another example is workers' compensation insurance. Some claims may not be reported until many years later (such as a disease brought about by occupational conditions), but would require medical payments throughout the rest of a person's life.
While auto insurance claims only involve predictions over the twelve months of the policy period, reserving for worker compensation requires the tools to predict expected claims payments decades into the future based on the information available today.
When it comes to your loyalty program, this is just the type of information you need.
Actuaries have the toolbox for making long-term predictions
Fortune-tellers have crystal balls; actuaries have numbers, data, and statistics. Using statistical analysis, they can evaluate the probability of future events occurring and plan accordingly.
In the context of loyalty programs, cash flow (amount, timing) is the primary metric examined. Other metrics can include:
- Customer lifetime value (CLV)
- Customer future value (CFV)
- Customer potential value (CPV)
- Ultimate redemption rate (URR)
- Cost-per-point (CPP)
- Breakage
Forecasting over the long term is required for insurance companies, and can be highly beneficial for loyalty programs. Let's walk through the benefits to long-term forecasting with actuaries.
9 reasons why actuarial insight is useful for loyalty programs
1. Helps predict loyalty program liability
When points are issued, companies must defer revenue for the eventual cost of redemption. This is required by the financial reporting standards ASC 606/IFRS 15. The challenge in estimating liability, however, lies in the number of variables associated with estimating redemption costs.
Actuaries can predict what percentage of points will ultimately be redeemed, when those redemptions will occur, and how much those redemptions will cost the company. After all, points can be redeemed many years (potentially decades) after they are earned. The best estimates require long-term forecasts.
Adding to this challenge is the reality that loyalty programs are dynamic, constantly evolving entities:
- Members change their behavior frequently
- Members are becoming increasingly sophisticated thanks to the availability of online resources on loyalty programs
- Programs frequently offer deals and marketing campaigns
- Programs occasionally change program structure – e.g., expiration rules, award charts, accrual charts, etc.
Liabilities can be very large, in the hundreds of millions or even billions of dollars – particularly among frequent flyer and hospitality programs. At this scale, CFOs and auditors require proper due diligence to validate liability estimates. Actuarial opinions provide formal documentation from a credentialed actuary stating their professional opinion on the booked program liability. This provides CFO, auditors, and other stakeholders proof of the due diligence.
2. Quantifies customer lifetime value (CLV)
While liability is important, the whole point of a loyalty program is to increase customer brand loyalty. (i.e., customers should keep coming back to your company to make purchases). The best way to quantify the value of this long-term loyalty is a metric known as customer lifetime value (CLV). CLV predicts the profits that a given member is expected to generate over his lifetime in the program.
As we've discussed in previous articles, CLV and its family of metrics are the most important KPIs to link program management to economic value creation. In fact, you can even use these metrics to proactively create economic value (you can review how to calculate customer lifetime value here).
Properly calculating CLV is critical to loyalty program profitability and requires the ability to predict over long horizons. As you'll see, CLV and its many applications will unify the rest of this section.
3. Identifies high value members
Simply put, members with high CLV are likely to spend more money with your company. Targeting them with special offers will make them feel appreciated and inspire even more purchases, maximizing the likelihood of unlocking their expected future profits.
4. Drives incremental value by targeting high potential members
With just a small nudge, high potential members will likely increase their program participation, and, as a result, their CLV. A well-executed, targeted incentive strategy can grow the future economic value of your loyalty program.
5. Targets at-risks members with decreasing CLV
On the flip side, loyalty programs can use CLV as a defensive strategy. By identifying leading indicators of declining participation, programs can be designed to proactively prevent loss of economic value.
6. Provides enhanced customer service based on CLV
CLV quantifies the value of each customer. This indispensable information enables customer service agents to focus their efforts and spend more time and energy on resolving issues for those members most valuable to the organization. Note that this doesn't mean that members with a low CLV don't matter; rather, it focuses efforts on providing services that will keep your most loyal customers happy.
7. Optimizes new member acquisition based on value rather than cost
Well-run loyalty programs can become an appreciating asset for any company. Examining CLV as a growth value metric ensures that every dollar spent on acquisition is spent in a way that maximizes economic value, rather than minimizing cost.
8. Optimizes program design
Actuaries conduct scenario testing to forecast the impact of various program changes on CLV. By quantifying how these changes impact CLV, program managers can identify and implement the changes that drive the most growth. This ensures that program structure changes are focused on maximizing economic value.
9. Aligns marketing and finance around a common set of KPIs to measure economic value creation
Miscommunication and lack of alignment leads to lost time in business. Aligning KPIs across organizations prevents organizational conflict. Using the shared language of CLV, organizations can increase the speed by which they make decisions — a competitive advantage in today's fast paced world.
Why a data science team isn't enough
In today's digital world, data analysis is a highly sought after skill. Wherever there's a large amount of data, data scientists are needed to plug and chug through significant statistical analyses, searching for patterns and identifying solutions.
At first glance, data scientists share many skills with actuaries. Both require business intelligence and data analysis skills. And the end result of data analysis and actuarial insight is a best-fit solution to an ambiguous problem.
However, these professions are not interchangeable.
Data scientists lack domain knowledge
Data scientists know how to build models to make predictions, but not necessarily predictions over long horizons, which is where actuaries excel. Actuaries are also subject to rigorous formal training and credentialing in their domain of focus – whether that be insurance, finance, investments, etc.
We can't overstate the value of domain knowledge. Consider this example:
Suppose you need brain surgery. Are you going ask your family doctor to do the operation, or a trained brain surgeon? Both are very smart, both are trained in the medical profession, and both probably have the aptitude to do it. But only the brain surgeon has the training, experience, and specific domain knowledge required to do the job correctly.
Data scientists and actuaries are similar: both are smart, both are trained in applied statistics, and both have the aptitude to learn to predict over long horizons; but only actuaries have the explicit training, experience, and specific domain knowledge required to do the job right.
What makes an ideal loyalty program actuary?
Traditional actuarial models focus on making predictions in aggregate. But the value of CLV lies in its ability to make predictions at the individual member level.
Within actuarial consulting there is even more niche domain knowledge required to properly apply actuarial theory to loyalty programs. The most effective professionals combine predictive modeling with expert financial reporting knowledge.
Individual predictive modeling
Loyalty program actuaries need to know how to leverage today's technology to build predictive models. Traditional actuarial methods were developed decades ago, well before the availability of today's big data and advanced predictive modeling capabilities.
While the basic actuarial concepts underlying traditional methods remain applicable to today's loyalty programs, the methods themselves need to be redesigned to leverage modern technology and allow for individual member-level predictions. Therefore, you need an actuary with strong predictive modeling skills and experience using modern technology to solve problems.
Financial reporting expertise
Given a loyalty program's complex financial reporting requirements, you not only need an actuary with predictive modeling skills, but also one with a strong understanding of the business dynamics and financial reporting regulations surrounding loyalty programs.
ASC 606/IFRS 15 have introduced updated revenue recognition standards and a fresh focus on loyalty program accounting practices. Your actuary needs to know how to properly identify performance obligations when accounting for loyalty programs.
In summary, the ideal loyalty program actuary is adept at predicting behavior over the long term, can build predictive models at the individual member level, and has a nuanced understanding of business dynamics and the financial reporting environment. This combination enables actuaries and loyalty program leaders to translate predictions into real business insight and loyalty program value.
Optimize your program with the right actuary
Loyalty programs generate more than a significant amount of data. Between engagement metrics, membership statistics and financial results, programs need someone with the skills to discern the meaning amidst all the noise. And while it's easy enough to turn to any data scientist or actuary, to get the most actionable insight from your data, it's best to trust those with deep domain expertise.
Imagine if each year at tax season, you hired an average accountant with limited personal tax experience. Would you really expect him or her to optimize your taxes and bring you the biggest refund?
The odds are that you would sleep better at night (and receive the biggest bang for your buck) if you hired a professional tax accountant with years of experience.
An ideal actuary can provide just as much peace of mind — and financial return — to your loyalty program. Select an expert who understands how to model your loyalty program liability, the nuances of customer lifetime value, and the fundamentals of optimizing program ROI.
Loyalty programs are one of the most effective ways of driving long-term customer value and ensuring the continued success of your business. When it comes to your program's financial management, why cut corners?


text:
Ever wondered if Second Life will run under Ubuntu64 (64-bit Ubuntu)?
Check this:
Oh yes, that is Second Life running on Ubuntu64.
Download the client here: SecondLife for GNU/Linux
Get this file: "Linux i686: 184.108.40.206 (Alpha) (50 MB)"
I know, it says i686 which refers to 32-bit but trust me it will run.
And no need to install, just extract it anywhere you want and run the secondlife file located in the extracted folder.
See you in-game!
SLname: Yarso Loire
Is a bibliophile and technophile other than being an early adopter, an avid gamer, anime otaku, trekker, and photographer. He is an advocate of "Free Culture", "Open Knowledge", "Creative Commons", "Free/Libre Open-Source Software", and the "Fediverse" (federated social-network).
His first online project was in 1998 when he launched the unofficial website for Ansalon MUD (a text-based online game) and his own community forums Laibcoms.Community. Today, he owns a variety of online properties and help others establish their online presence.
Confirm his identity through Keybase, and learn more by reading his central bio. Yuki's hubsite, Tomes of Knowledge, is located at jcsesecuneta.com.
Second Life in Ubuntu64 by gameshogun™ 🎲 is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License. Permissions beyond the scope of this license may be available at Legal Notice.


text:
Sleep Innovations 14-Inch Memory Foam Mattress is perfect for distributing body weight evenly that leads to pressure points relief to protect you from tossing and turning.
This 14-inch Sleep Innovations mattress is designed to keep its actual form without any problems of flipping or rotating. The mattress is designed of 3 layers, top, middle and bottom layers. Top layer is of 2.5-inch open cell memory foam to offer utmost cradling comfort.
Middle layer is of 2.5-inch air channel foam to provide improved airflow and personalized support. The bottom layer is made of 9-inch support foam for greater durability. Sleep Innovations super comfort mattress is designed to give you the greatest night sleep ever.
Sleep Innovations 14-Inch Memory Foam Mattress features combined 5-inches of premium memory foam and air channel foam to provide the best comfort and customized sleeping experience. This Sleep Innovations brand is dedicated towards providing the ultimate comfort and therapeutic support at a great value. All the memory foam it uses is made in the USA to provide the highest comfort level at the lowest price. Read more…


text: Really excited to announce that PUCKLAND, a docu-series airing in late March on NBC Sports will be using five Zambonis tunes throughout the series. Puckland is about the journey of minor league players trying to get to the NHL and also about the challenges to get professional hockey back in Portland, ME. Check your local listings.


text:
The global enteral feeding devices market is filled with various players on both, regional and international level. As a result of this, the global enteral feeding devices market is anticipated to be very fragmented. To name a few, Moog, Inc., Kimberly Clark, Covidien plc, Applied Medical Technology, Inc., CONMED Corporation, Amsino International, Inc., Boston Scientific are among the most dominant firms in the enteral feeding devices market. These firms account for an edge over their competitors to score their current leading positions. These organizations working consistently on coming up with efficient strategies to achieve the position, including mergers, acquisitions, and collaborations. They are also working upon regressive and intelligent promotional activities.
According to a recent report by Transparency Market Research, the global market for enteral feeding devices is prognosticated to expand at a decent pace with a steady CAGR. The market is expected to grow around worth of US$3.1 bn by the end of 2019.
These days, there are several types of enteral feeding devices present in the market. Among these, the enteral feeding pumps segment is anticipated to experience highest sales in the market. This is majorly due to very negligible amount of risk and complication, increase in their usage in home care settings, and portability. As per the geography, Europe is a leading region in the global market for enteral feeding devices. This is due to significant rise in population in the region in past few years.
Constant Efforts from Manufacturers to Fuel Market Growth
The global enteral feeding devices market is witnessing a major shift owing to the gadgets fast outstripping demand for parenteral gadgets. This is majorly due to the presence of better technologies for example low scale gastrostomy. Moreover, enteral feeding sets and tubes are very affordable, as well. This is another factor propelling the sales of enteral feeding devices market.
Nevertheless, the global enteral feeding devices market is also drawing attention because of the rigorous efforts by the key manufacturers to come up with improved products, with even less complications and side effects. The launch of balloon inflated gastrostomy tubes that allow medicines and food to be directly administered to the patient's stomach, without requiring them to swallow, bite or chew has supported the uptake in the market.
Rise in number of elderly people all over the world coupled with the growing preference for home care enteral therapy has fueled the global enteral feeding devices market. Rise in cases of premature births which needs feeding of the infants through enteral devices are contributing in the growth of the market as well.
Inconvenient Reimbursement Policies to Hamper Overall Market Growth
However, on the flipside, the market still needs to work upon the complexities arising from enteral nutrition delivery, such as non-obstructive intestinal ischemia, diarrhea, and gastrointestinal complications. Inconvenient reimbursement policies regarding enteral feeding devices is as well hindering demand globally to a large extent. Nevertheless, rising cases of cancers and gastrointestinal disorders are foreseen to help eliminate the setbacks by supporting demand long time.
This analysis is based on a recent market research report by Transparency Market Research, titled, "Enteral Feeding Devices Market (Enteral Feeding Tubes, Enteral Feeding Pumps, Administration Reservoir, Giving Sets and Enteral Syringes) - Global Industry Analysis, Size, Share, Growth, Trends and Forecast 2013 – 2019."
Key Takeaways:
- Enteral feeding sets and tubes are very affordable, as well. This is another factor propelling the sales of enteral feeding devices market.
- Rise in number of elderly people all over the world coupled with the growing preference for home care enteral therapy has fueled the global enteral feeding devices market.
The global enteral feeding devices market is segmented into the following categories:
- Enteral Feeding Devices Market, by Product Types
- Enteral Feeding Tubes
- Nasoenteric Tubes
- Naso-gastric
- Naso-duodenal
- Naso-jejunal
- Oroenteric Tubes
- Oro-gastric
- Oro-duodenal
- Oro-jejunal
- Enterostomy Tubes
- Low Profile Gastrostomy
- Nasoenteric Tubes
- Enteral Feeding Pumps
- Administration Reservoir
- Giving Sets
- Enteral Syringes
- Enteral Feeding Tubes
- Enteral Feeding Devices Market, by Stages
- Pediatrics
- Adult
- Enteral Feeding Devices Market, by Geography
- North America
- Europe
- Asia-Pacific
- Rest of the World (RoW)
About Us:
Transparency Market Research is a next-generation market intelligence provider, offering fact-based solutions to business leaders, consultants, and strategy professionals.
Our reports are single-point solutions for businesses to grow, evolve, and mature. Our real-time data collection methods along with ability to track more than one million high growth niche products are aligned with your aims. The detailed and proprietary statistical models used by our analysts offer insights for making right decision in the shortest span of time. For organizations that require specific but comprehensive information we offer customized solutions through adhoc reports. These requests are delivered with the perfect combination of right sense of fact-oriented problem solving methodologies and leveraging existing data repositories.
TMR believes that unison of solutions for clients-specific problems with right methodology of research is the key to help enterprises reach right decision.
Transparency Market Research
90 State Street, Suite 700
Albany, NY 12207
Tel: +1-518-618-1030
USA - Canada Toll Free: 866-552-3453
Email: email@example.com
Website: https://www.transparencymarketresearch.com
7 of 10 large enterprizes view our ToC to take the right decision.View Report ToC


text:
Boost Followers
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In addition to "Would You Rather," I also got the chance to ask the Man from U.N.C.L.E. cast about a few specific details in the film. The Guy Ritchie-directed movie stars Henry Cavill as Napoleon Solo and Armie Hammer as Ilya Kuryakin. Solo is with the CIA and Kuryakin is a KGB agent but they've got to team up and work together in order to stop Victoria Vinciguerra (Elizabeth Debicki) from getting her hands on nuclear weaponry.
Check out what Cavill, Hammer, Debicki and Alicia Vikander told me about the nauseating experience shooting on a boat, Vikander and Hammer's dancing/wrestling scene, the stunning fashion in the movie and much more in the video interview below. The Man from U.N.C.L.E. hits theaters nationwide on August 14th and I can't recommend it enough.
Here's the official synopsis for The Man from U.N.C.L.E.:
Set against the backdrop of the early 1960s, at the height of the Cold War, The Man from U.N.C.L.E. centers on CIA agent Solo and KGB agent Kuryakin. Forced to put aside longstanding hostilities, the two team up on a joint mission to stop a mysterious international criminal organization, which is bent on destabilizing the fragile balance of power through the proliferation of nuclear weapons and technology. The duo's only lead is the daughter of a vanished German scientist, who is the key to infiltrating the criminal organization, and they must race against time to find him and prevent a worldwide catastrophe.


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I woke up to the sound of snickering and laughter coming from the other side of the bedroom door. My head was spinning, body trembling, and heart pounding. I closed my eyes again and tried to convince myself it was just a really bad dream. This kind of thing doesn't happen to girls like me…
And what kind of girl would that be? Let me give you some context. A girl who just gets dressed up for a night of fun… who doesn't think twice about trying to look beautiful to go to a party…girls who value who they are on the inside and out…
But, sadly, it did happen. And it happens more often than you might imagine–to beautiful girls who, like me, who are trying to be the best they can be.
Beautiful girls. How does the word beauty play into my story? Let's take a step back. What is the definition of beauty? Personally, when I think of beauty, I imagine snapshots of creation. Sunrises that paint the morning into existence, sunsets of neon colors, northern lights twinkling in a sky full of constellations, white sands that stretch on for miles, bright blue Caribbean waters, mountains that unabashedly reach for the heavens, the list is limitless. But in pragmatic terms, beauty is defined in the Merriam-Webster dictionary as "the quality or aggregate of qualities in a person or thing that gives pleasure to the senses or pleasurably exalts the mind or spirit." It goes on to include these words "especially: a beautiful woman."
That last part, did you catch it? Why do you suppose the dictionary includes a woman under the definition of beautiful? There's no doubt the snapshots of creation exalt my spirit, but Eve was undoubtedly the crowning jewel of all creation. Ladies, we are God's pièce de résistance (masterpiece). What a gift we've been given!
From Cleopatra to Joan of Arc, men have crossed the seas and fought many a war for a beautiful woman. There is undoubtedly a mighty power in the beauty a woman holds that can cause a man to cross seas and fight wars. But, there is a flip side to this. Usher in a fallen world, and with that fall came a thirst for power over and possession of beautiful things. An incredible gift so easily twisted, distorted, idolized, and stripped apart from God's holy intentions. What God intended for good, man can use for evil.
The depth and danger of such ungodliness came crashing into my own little world my sophomore year of college. I was best described as a conservatively-bred, partially-sheltered, naïve-in-the-collegiate-athletic culture, far-from-her-roots, nineteen-year-old. Being young and naïve surely helped fuel the elation I felt upon being invited to a joint birthday party for a teammate and her guy-friend at a high-rise penthouse in my new home and among my new friends in Miami. I couldn't wait to get all dolled up to join in on the fun. As I stepped out for an expectant evening, I felt what Eve and millions of women for centuries have felt–beautiful.
But beauty turned to ashes that night in the blink of a mascara-smeared eye. I awoke to the realization that I had been gang raped by four party goers; one of them was the birthday boy himself. I was crushed. I grieved what was stolen from me. I wanted nothing more than to hide from the world. I convinced myself this "gift" of being a woman was actually a curse and I was so angry with God. I strayed away from my Heavenly Father and from my true self. I built a wall in my heart so high and I pushed away anyone and everyone that came too close. I went numb.
"What makes you vulnerable, makes you beautiful." – Brené Brown
Ladies, sometimes it feels safer to simply hide who you are–a beautiful woman made by God–because with womanly beauty also comes vulnerability. And vulnerability means "being open to attack or damage; being capable of physical or emotional damage." I did NOT like that feeling.
But, then something else happened. I was sporadically attending a church down near my college and on one of the Sundays I attended, the sermon was centered on 1 Corinthians 12:9-10.
"But He said to me, My grace is sufficient for you, for my power is made perfect in weakness. Therefore I will boast all the more gladly of my weaknesses, so that the power of Christ may rest upon me. For the sake of Christ, then, I am content with weaknesses, insults, hardships, persecutions, and calamities. For when I am weak, then I am strong."
Lightbulb. And gut-check.
I was trying so hard to rely on my own strength and to come up with my own answers as to why this happened to be. I tried to be tough. And, with that, I had all but lost who God made me to be. Sure, on the exterior I still looked the part, and in front of close friends and family I acted the part as well, but my light inside had been snuffed out. That day, at church, I tapped out. It was time to stop relying on my own strength and trust in the one who made me. The one who calls me beautiful and the one who knows what power I have in His name and in His image.
Matthew 11: 28-30 "Come to me, all you who are weary and burdened, and I will give you rest. Take my yoke upon you and learn from me, for I am gentle and humble in heart, and you will find rest for your souls. For my yoke is easy and my burden is light."
My soul needed rest. Once I laid my hurt and anger at His feet, I felt like my own feet began to float. I died to myself that day; and like a phoenix I rose from those ashes. That, my friends, is a beautiful thing. I felt the spark in my soul begin to rekindle. I felt capable of moving forward and being the beautiful, but humble; strong, but empathetic; fiery, but sweet woman the God of the Universe intended for me to be. And no man can ever take that away. I realized that evil exists, but God is greater. Flip! Genesis 50:20 "As for you, you meant evil against me, but God meant it for good, to bring it about that many peopleshould be kept alive, as they are today."
Yes, history has shown us that there is power in physical beauty. It has also shown us that many have used that power in the wrong way. With great power comes great responsibility. The Bible makes it clear that true beauty is more than physical beauty. Your beauty is not solely defined by your outward appearance. When I chose to become emotionally numb, distant, and turn the lights off, I lost the true power that comes with beauty. My ability to nurture, love, comfort, empathize, guide and encourage those around me disappeared. Here's a hard truth: it is possible to look beautiful on the outside while being the opposite on the inside. I am here to urge you to resist the cocoon! Don't hide away and deprive those that need you to be your best self of your gifts and talents.
Proverbs 31:30 "Charm is deceptive, and beauty is fleeting; but a woman who fears the LORD is to be praised."
Embrace that you are a beautiful woman, inside and out, created by a loving father. Know what power you hold and use it for good. When you rest in the comfort of the Lord Almighty, when you run to your Heavenly Father and choose to be all that He intended for you, you are a force to be reckoned with and no enemy shall prevail against you. You will change more lives than you could have ever imagined. When we long for a life without difficulties remember this, Oak trees are made strong in contrary winds and diamonds are made beautiful under pressure.
My story has a redemptive ending that only the God of the Universe could have scripted. Shortly after I was convicted that bunkering down wasn't what God desired for me, I met the man who would become my husband, Zach. God knew exactly what kind of man I needed. Zach came into my life with a sledge hammer in one hand and tender touch in the other. He tore down the wall I had built around me. This August will be our fourth anniversary and we have two beautiful little girls.
As Romans 8:28 says "And we know that all things work together for good to those who love God, to those who are called according to His purpose." My story is woven into the grand narrative of our savior and because I surrendered the hurt and heartache to Him, I am free and fully able to guide others to the grace and peace that can be found nowhere else. Let your light shine, be beautiful inside and out, know the power you have and use it constructively to lift others up. Remember, you are God's masterpiece! And God doesn't make mistakes.
One of my absolute favorite quotes sums it up so well:
"Our deepest fear is not that we are inadequate. Our deepest fear is that we are powerful beyond measure. It is our light, not our darkness, that most frightens us. Your playing small does not serve the world. There is nothing enlightened about shrinking so that other people won't feel insecure around you. We are all meant to shine as children do. It's not just in some of us; it is in everyone. And as we let our own lights shine, we unconsciously give other people permission to do the same. As we are liberated from our own fear, our presence automatically liberates others."
-Marianne Williamson
Andi lives in the Panhandle of Florida with her husband and two girls Maison and Blake-leigh. Being the wife of an active duty Pararescueman (AKA Guardian Angel) comes with its unique set of challenges, but one thing for certain is that life is never boring. She currently works as a Personal Trainer and has made it her mission to remind women whose they are and why they were created to thrive, not merely survive. She uses her testimony to positively impact others and coach them to reach their God-given potential both physically and spiritually.
Follow Andi on socials media!
Instagram @andi_elizabeth21


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This is a picture of me last May (okay not quite a year, but I couldn't find any others). My hair is a lot shorter now and I'm a mother of 3 now, instead of just 2. Besides being (almost) a year older, I'm also a lot wiser than I was back then. It's not like a lot of time has passed, but it sure does feel like it. And then again, that picture feels like it was taken yesterday.
6 days ago


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Ammunition Market to Showcase Vigorous Demand during the Period until 2025
"Countries across the globe are facing significant concerns related to terrorism, civil wars and armed conflicts between law enforcement agencies and anti-national groups. Numerous governments are striving extensively to tackle such situation peacefully. However, in certain situations, harsh steps are needed to be taken such as use of fire arms in favor of national security. Current scenario has created a continuous demand for ammunition to tackle the aforementioned situation in most of the countries. In the age of arms race, every country allot a considerable budget for arms and ammunition for their country's security. This has elevated the ammunition market and is expected to grow further in the following years.
Get Sample PDF for Professional Insights: https://www.trendsmarketresearch.com/report/sample/3617
On the basis of end use, the market is segmented into civil and defense. The use of ammunition in civil sector is further segmented into self-defense, sports and hunting, and law enforcement. Market growth is influenced by military advancements in Russia, and increasing participation of civilians in hunting, shooting, and sports; particularly in North America and Europe.
In North America, U.S. is a pioneer in research and development (R&D) of ammunition industry. In Europe, the market is dominated by manufacturing of small arms and weapons industry. In the Asia Pacific region, the demand for ammunition is expected to be driven by the increase in cross-border issues and increase in the defense expenditure on account of rising economy. Countries such as China, India, Korea, South Africa, and Saudi Arabia are expected to boost the market growth.
Global Ammunition Market to Earn an Around US$XXMillion by the Completion Of 2025
Demilitarization activities by developed economies in North America and Europe, coupled with high costs associated with research and development is expected to hinder the market demand from the law enforcement and military sectors. However, these factors are expected to be offset by the significant growth in the Asia Pacific and Middle East regions.
Key players in the global ammunition market include Orbital ATK Inc. (U.S.), Olin Corporation (U.S.), General Dynamics Corporation (U.S.), BAE Systems (U.K.), Remington Arms Company, LLC (U.S.), and Nammo AS (Norway), among others.
Get Complete ToC with Tables and Figures at : https://www.trendsmarketresearch.com/report/requesttoc/3617
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Press Release
The Canalta Hotels AJHL Player of the Week is Brennan Saulnier of the Fort McMurray Oil Barons. Saulnier scored 3 goals, 3 assists and 6 points in three games to close out the month of January 2014.
Brennan Saulnier opened the week with a hat trick and a 4 point game in an Oil Barons win over the Bonnyville Pontiacs on January 28th. The hat trick was a first for Saulnier in the AJHL. Saulnier was selected as the Pro Hockey Life Player of the Game in two consecutive games – the first for his performance on January 28th and the second on January 29th against the Brooks Bandits. Brennan wrapped up the week with an assist on the game winner in a victory over the Calgary Mustangs on February 1st.
The 20 year-old forward from Halifax, NS is in his first AJHL season with the Oil Barons. Saulnier and the MOB hold second place in the AJHL North Division Standings with 81 points and 11 regular season games remaining.
The Alberta Junior Hockey League is proud to partner with Canalta Hotels to continue recognition of the AJHL Player of the Week for the 2013-2014 AJHL Season.
(Nathan can be reached at email@example.com)
31 Teams in 31 Days - Los Angeles Kings
21 hours ago


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Do you think our early ancestors wore half-length or full-length orthotic shoe inserts? Would their video-recorded gait assessments have revealed more under or over-pronators in the general population?
If we try to imagine how life was back in the day of the caveman we could safely assume that his ability to run both short sprints and long distances was necessary for surviving in a wild environment. Without the ability to track food or flee from dangerous predators the human evolution may not have been possible. What we can also be sure of is the caveman did not run in a pair of Asics trainers with rear-foot gel-cushioning outer soles structured for either a neutral or over-pronating gait . Nope, our early ancestors would have been running barefoot.
Many thousands of years later and despite our advances in footwear design and technology there is a 'barefoot running' revolution gaining traction in some circles. The idea is that we as a species have spent more time running barefoot than we have in our fancy high-tech shoes and therefore we are more physiologically suited to running barefoot.


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|Emmanuel Lécharny <elecha...@gmail.com>
|Bulk Load
|Fri, 30 May 2014 09:35:12 GMT
Hi guys, Kiran's work on bulk load is making huge progress. On my computer, I was able to load 30 000 entries in around 16s, and the last modification he is working on (ie, make teh LdifReader schema aware) will make it going down to 15s. This is 2000 entries added per second, with all the indexes created (10 of them). Here is the result of a run : Loading schema using JarLdifSchemaLoader Sorting the LDIF data... Using dc=example,dc=com as the partition's root DN Completed sorting, total number of entries 30002, time taken : 2462ms Creating partition..., time taken : 174ms Building master table...replacing old offset 17920 of the BTree master with 70266368 , time taken : 10145ms Building RDN index.replacing old offset 13312 of the BTree 220.127.116.11.4.1.18060.0.4.1.2.50_forward with 92644864 replacing old offset 14848 of the BTree 188.8.131.52.4.1.18060.0.4.1.2.50_reverse with 113890304 , time taken : 2223ms Clearing the sorted DN set. Building index objectClassreplacing old offset 92649472 of the BTree 6f5011bf-c0cb-46fa-ba92-4834b0a1124f with 115933184 replacing old offset 115934720 of the BTree 6c8b2473-abf9-4ca5-bb6c-a6ab99566026 with 117976576 replacing old offset 117978112 of the BTree e6b700bc-9c37-4436-8f18-ff7a05323f30 with 120019968 replacing old offset 10240 of the BTree 126.96.36.199_forward with 120020992 , time taken : 252ms Building index entryCSNreplacing old offset 1536 of the BTree 220.127.116.11.4.1.4203.666.1.7_forward with 124040192 , time taken : 142ms Building index administrativeRole, time taken : 13ms Building index apacheOneAlias, time taken : 18ms Building index apacheSubAlias, time taken : 12ms Building index apacheAlias, time taken : 10ms Building presence index...replacing old offset 11776 of the BTree 18.104.22.168.4.1.18060.0.4.1.2.3_forward with 120022528 , time taken : 15ms Patition building complete in 16167ms So the most expensive operations are the MasterTable creation (2/3rd of the time), the RDN index creation, and the entry sorting. All those three operations are extensivly manipulating DN. We can greatly improve the performance by speeding up the DN parsing here. How ? Every DN will be read as a String, then parsed fully. This is most certainly useless, as most of the DN will have the same parent. We can keep in cache the parents, and get rid of them sparing the cost of parsing again, and agaiun, and again, those DN. For instance, if we read cn=test,dc=example,dc=com, we are very likely to hava already parsed dc=example,dc=com. This can easily be checked by looking if the current DN ends with dc=example,dc=com. If this is the case, we can then split teh DN in two parts, the child part and the parent part, and only parse the child part. I think that could save a lot of processing time. Now, we have to be careful, because we can't do that for evey single RDN we already have parsed. At least, checking for the naming context could help. Btw, I see that we are parsing the DN 181 000 times (15% of the spent time). We can most certainly save most of those parsings. Here is where we do parse the DN : - when we parse the LdifEntry (30 000 times) - in the FastLdifReader ( 30 000 times) - when we parse the creratorsName attribute (it's a DN, and we do check its syntax) ( 30 000 times) And some other places. Here is the stack trace : org.apache.directory.api.ldap.model.name.FastDnParser.parseDn(String, List) 4976 181712 org.apache.directory.api.ldap.model.name.Dn.parseInternal(String, List) org.apache.directory.api.ldap.model.name.Dn.isValid(String) 3160 90856 org.apache.directory.api.ldap.model.schema.syntaxCheckers.DnSyntaxChecker.isValidSyntax(Object) 1588 30002 org.apache.directory.api.ldap.model.ldif.LdifReader.parseDn(String) 1571 60854 org.apache.directory.api.ldap.model.ldif.LdifReader.parseEntry() 844 30852 org.apache.directory.api.ldap.model.ldif.LdifReader.parseLdif(BufferedReader) 743 30002 org.apache.directory.api.ldap.model.name.Dn.<init>(SchemaManager, String[]) 1816 90856 org.apache.directory.api.ldap.model.name.Dn.<init>(String[]) 1475 60854 org.apache.directory.api.ldap.model.name.Dn.<init>(String[]) 1475 60854 org.apache.directory.mavibot.btree.FastLdifReader.parseDnAlone() 698 30002 org.apache.directory.api.ldap.model.schema.normalizers.DnNormalizer.normalize(String) 341 30002 So there are some room for imprtovement in this area :-)
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