Difference In Colony-forming Units Research Articles

In Vitro and In Vivo Testing of Microbe Growth on Antimicrobial Nursing Scrubs.

Around 5% to 10% of hospitalized patients develop a hospital-acquired infection (HAI). Scrubs are a potential vector of HAIs. To compare the antimicrobial characteristics of scrubs with and without an antimicrobial fabric coating, as tested in the laboratory (in vitro) and hospital (in vivo) environments. Two protocols were conducted to address the purpose. The in vitro protocol was a laboratory study that involved observing the microbe growth after inoculating coated and uncoated scrub fabric swatches with S. aureus and then processing them in moist and dry environments. The in vivo protocol was a clinical trial that measured microbe growth on coated and uncoated scrubs prior to and following nursing staff completing a 12-hr shift on an acute care unit, as measured by colony forming units (CFUs). For high-humidity environments, the in vitro study indicated that swatches treated with an antimicrobial coating exhibited minimal microbe growth, while untreated swatches exhibited significant microbe growth. For low-humidity environments, coated and uncoated swatches were all found to exhibit minimal microbe growth. In the in vivo study, the CFUs increased on scrubs worn by nurses over a 12-hr shift with no significant difference in CFUs for coated and uncoated scrubs. For bacteria in a warm and moist environment, the antimicrobial coating was found to be important for inhibiting growth. For bacteria in a warm and dry environment, both coated and uncoated fabrics performed similarly as measured at 24 hr, with minimal bacterial growth observed. In a hospital environment, microbe growth was observed, but no significant difference was detected when comparing coated and uncoated scrubs. This may have been due to the short time between exposure and culturing the scrubs for analysis immediately at the end of the shift not allowing for enough time to kill or inhibit growth. Contact time between the bacteria and scrub fabric (coated or uncoated) in the in vivo study more directly correlated with the 0-hr observations for the in vitro study, suggesting that the ineffectiveness of the treated scrubs in the clinical results may be due in part to short residence times before collection.

Effectiveness of Negative Pressure Wound Therapy During Aeromedical Evacuation Following Soft Tissue Injury and Infection.

Negative pressure wound therapy (NPWT) is utilized early after soft tissue injury to promote tissue granulation and wound contraction. Early post-injury transfers via aeromedical evacuation (AE) to definitive care centers may actually induce wound bacterial proliferation. However, the effectiveness of NPWT or instillation NPWT in limiting bacterial proliferation during post-injury AE has not been studied. We hypothesized that instillation NPWT during simulated AE would decrease bacterial colonization within simple and complex soft tissue wounds. The porcine models were anesthetized before any experiments. For the simple tissue wound model, two 4-cm dorsal wounds were created in 34.9 ± 0.6 kg pigs and were inoculated with Acinetobacter baumannii (AB) or Staphylococcus aureus 24 hours before a 4-hour simulated AE or ground control. During AE, animals were randomized to one of the five groups: wet-to-dry (WTD) dressing, NPWT, instillation NPWT with normal saline (NS-NPWT), instillation NPWT with Normosol-R® (NM-NPWT), and RX-4-NPWT with the RX-4 system. For the complex musculoskeletal wound, hind-limb wounds in the skin, subcutaneous tissue, peroneus tertius muscle, and tibia were created and inoculated with AB 24 hours before simulated AE with WTD or RX-4-NPWT dressings. Blood samples were collected at baseline, pre-flight, and 72 hours post-flight for inflammatory cytokines interleukin (IL)-1β, IL-6, IL-8 and tumor necrosis factor alpha. Wound biopsies were obtained at 24 hours and 72 hours post-flight, and the bacteria were quantified. Vital signs were measured continuously during simulated AE and at each wound reassessment. No significant differences in hemodynamics or serum cytokines were noted between ground or simulated flight groups or over time in either wound model. Simulated AE alone did not affect bacterial proliferation compared to ground controls. The simple tissue wound arm demonstrated a significant decrease in Staphylococcus aureus and AB colony-forming units at 72 hours after simulated AE using RX-4-NPWT. NS-NPWT during AE more effectively prevented bacterial proliferation than the WTD dressing. There was no difference in colony-forming units among the various treatment groups at the ground level. The hypoxic, hypobaric environment of AE did not independently affect the bacterial growth after simple tissue wound or complex musculoskeletal wound. RX-4-NPWT provided the most effective bacterial reduction following simulated AE, followed by NS-NPWT. Future research will be necessary to determine ideal instillation fluids, negative pressure settings, and dressing change frequency before and during AE.

Effect of fenestrated sterile drape and face mask on bacterial dispersion toward the periocular area during intravitreal injection

This experimental crossover study was performed to investigate whether fenestrated surgical drapes (covering the nose and mouth but with an opening over the periorbital area) with or without patients’ surgical face masks increase periorbital bacterial dispersion during simulated intravitreal injection conditions. Each of the 16 healthy volunteers performed 14 scenarios involving different mask and drape conditions in both silent and speaking situations. In each scenario, the subject lay down flat on the back with a blood agar plate being held at the inferior orbital rim perpendicular to the face to capture airflow from breathing/speaking. Another blood agar plate placed 50 cm away from the subject served as an experimental control. A total of 224 experiments were performed. Speaking situations significantly showed more colony forming units (CFUs) compared with their controls (P = 0.014). There were no significant differences in CFUs between wearing vs not wearing the masks (P = 0.887 for speaking and P = 0.219 for silent) and using vs not using the drapes (P = 0.941 for speaking and P = 0.687 for silent). Reusable and disposable drapes were also not significantly different (P = 1.00 for speaking and P = 0.625 for silent). Streptococcus spp., the oropharyngeal microbiota, were only cultivated from speaking scenarios. While refraining from speaking (for both practitioners and patients) is the mainstay of reducing bacterial dispersion and risks of post-injection endophthalmitis, the use of fenestrated surgical drapes or patients’ face masks did not significantly affect the amount of bacterial dispersion toward the periorbital area.

Clinically relevant pathogens on surfaces display differences in survival and transcriptomic response in relation to probiotic and traditional cleaning strategies

Indoor surfaces are paradoxically presumed to be both colonized by pathogens, necessitating disinfection, and “microbial wastelands.” In these resource-poor, dry environments, competition and decay are thought to be important drivers of microbial community composition. However, the relative contributions of these two processes have not been specifically evaluated. To bridge this knowledge gap, we used microcosms to evaluate whether interspecies interactions occur on surfaces. We combined transcriptomics and traditional microbiology techniques to investigate whether competition occurred between two clinically important pathogens, Acinetobacter baumannii and Klebsiella pneumoniae, and a probiotic cleaner containing a consortium of Bacillus species. Probiotic cleaning seeks to take advantage of ecological principles such as competitive exclusion, thus using benign microorganisms to inhibit viable pathogens, but there is limited evidence that competitive exclusion in fact occurs in environments of interest (i.e., indoor surfaces). Our results indicate that competition in this setting has a negligible impact on community composition but may influence the functions expressed by active organisms. Although Bacillus spp. remained viable on surfaces for an extended period of time after application, viable colony forming units (CFUs) of A. baumannii recovered following exposure to a chemical-based detergent with and without Bacillus spp. showed no statistical difference. Similarly, for K. pneumoniae, there were small statistical differences in CFUs between cleaning scenarios with or without Bacillus spp. in the chemical-based detergent. The transcriptome of A. baumannii with and without Bacillus spp. exposure shared a high degree of similarity in overall gene expression, but the transcriptome of K. pneumoniae differed in overall gene expression, including reduced response in genes related to antimicrobial resistance. Together, these results highlight the need to fully understand the underlying biological and ecological mechanisms for community assembly and function on indoor surfaces, as well as having practical implications for cleaning and disinfection strategies for infection prevention.

A Randomized Clinical Trial Comparing the Efficacy and Safety of 2% Versus 4% Chlorohexidine Gluconate for Surgical Preparation of the Vagina.

There have been no studies comparing 2% and 4% chlorhexidine gluconate (CHX) for vaginal surgical site preparation despite both commonly being used. The primary objective was to assess the noninferiority of 2% CHX versus 4% CHX to reduce bacterial contamination rates at 60 minutes after surgical preparation of the vagina. Secondary objectives were to assess differences in colony-forming units and the sensation of postoperative vaginal burning and pain. This is a single-blinded randomized controlled trial of women who underwent vaginal prolapse surgery. Study participants were randomized into 2 groups: 2% CHX versus 4% CHX. Two vaginal bacterial cultures were collected: (1) preoperatively before vaginal preparation and (2) intraoperatively at 60 minutes. A questionnaire on vaginal pain and burning was administered preoperatively and postoperatively. For our sample size, assuming that 2% CHX would have double the contamination rate of 4% CHX, 26 participants were needed per group to demonstrate noninferiority. Sixty-one women participated in the study. There were no differences in baseline demographics, length of surgery, or surgical procedures. For our primary outcome, the postpreparation contamination rates were 7% for 2% CHX versus 10% for 4% CHX, with a difference of 3% ( P = 0.52). This difference did meet the criteria for noninferiority. Secondary outcomes were not different between groups. Two percent CHX is noninferior to 4% CHX in reducing vaginal bacterial contamination at 60 minutes after vaginal surgical site preparation, with low rates of postpreparation contamination and vaginal discomfort.

The impact of Lactococcus lactis (probiotic nasal rinse) co-culture on growth of patient-derived strains of Pseudomonas aeruginosa.

The Lactococcus strain of bacteria has been introduced as a probiotic nasal rinse for alleged salubrious effects on the sinonasal bacterial microbiome. However, data regarding interactions with pathogenic bacteria within the sinuses are lacking. The purpose of this study is to assess the interaction between L. lactis and patient-derived Pseudomonas aeruginosa, an opportunistic pathogen in recalcitrant chronic rhinosinusitis (CRS). Commercially available probiotic suspension containing L. lactis W136 was grown in an anaerobic chamber and colonies were isolated. Colonies were co-cultured with patient-derived P. aeruginosa strains in the presence of porcine gastric mucin (mimicking human mucus) for 72 hours. P. aeruginosa cultures without L. lactis served as controls. Colony forming units (CFUs) were compared. Six P. aeruginosa isolates collected from 5 CRS patients (3 isolates from cystic fibrosis [CF], 1 mucoid strain) and laboratory strain PAO1 were co-cultured with L. lactis. There was no statistical difference in CFUs of 5 P. aeruginosa isolates grown with L. lactis compared to CFUs without presence of L. lactis. CFU counts were much higher when the mucoid strain was co-cultured with L. lactis (CFU+L.lactis = 1.9 × 108 ± 1.44 × 107, CFU-L.lactis = 1.3 × 108 ± 8.9 × 106, p = 0.01, n = 7). L. lactis suppressed the growth of 1 P. aeruginosa strain (CFU+L.lactis = 2.15 × 108 ± 2.9 × 107, CFU-L.lactis = 3.95 × 108 ± 4.8 × 106, p = 0.03, n = 7). L. lactis suppressed the growth of 1 patient P. aeruginosa isolate and induced growth of another (a mucoid strain) in in vitro co-culture setting in the presence of mucin. Further experiments are required to assess the underlying interactions between L. lactis and P. aeruginosa.

Structure-Property-Toxicity Relationships of Graphene Oxide: Role of Surface Chemistry on the Mechanisms of Interaction with Bacteria.

Graphene oxide (GO) is an antimicrobial agent with tunable surface chemistry. To identify the physicochemical determinants of GO's antimicrobial activity, we generated different modified Hummer's GO materials thermally annealed at 200, 500, or 800 °C (TGO200, TGO500, and TGO800, respectively) to modify the surface oxygen groups on the material. Plating assays show that as-received GO (ARGO) and TGO200, TGO500, and TGO800 reduce Escherichia coli viability by 50% (EC50) at 183, 143, 127, and 86 μg/mL, respectively, indicating higher bacterial toxicity as ARGO is reduced. To uncover the toxicity mechanism of GO, fluorescent dye-based assays were used to measure oxidative stress at the EC50. ARGO showed an increase in intracellular reactive oxygen species, measured as an increase in 2',7'-dichlorodihydrofluorescein diacetate fluorescence, whereas TGO500 and TGO800 induced an increase in the fluorescence of fluorescein diacetate (FDA) by 30 and 42%, suggesting a decrease in cell permeability. Because of a possible wrapping mechanism, plating assays after post-exposure sonication were performed to explain TGO's low oxidative response and high FDA levels. Results show no difference in colony-forming units, indicating that inhibition of cell growth is a result of the adsorption of bacterial cells on the GO material. By comparing different GO samples at their EC50, this study reveals that reduction of GO alters both the mechanisms of cellular interaction and the degree of toxicity to bacteria.

Assessment of sensitivity of selected Candida strains on antimicrobial photodynamic therapy using diode laser 635 nm and toluidine blue - In vitro research.

Photodynamic therapy is believed to be a promising treatment for Candida infections. This study evaluated the efficacy of antimicrobial photodynamic therapy (aPDT) using the 635 nm diode laser light and toluidine blue (TB) in the elimination of selected Candida species cultured on acrylic surface. 108 acrylic plates (Methyl Methacrylate Polymer, routinely used for the production of prosthetic dentures) were placed in three sterile Petri dishes and poured with prepared suspensions of Candida strains: C. albicans, C. glabrata, and C. krusei. After all procedures of fungi incubation, fungal biofilm was visible on the plates' surfaces. The acrylic plates were divided into nine study groups (B) and nine control groups (K) for further experiments. In the study groups, the acrylic plates with fungal biofilm were immersed in TB and afterwards laser irradiation was applicated with different exposure parameters (groups: B1 - 400 mW, 24 J/cm2, 30 s; B2 - 300 mW, 18 J/cm2, 30 s; B3 - 200 mW, 12 J/cm2, 30 s) separately for each Candida species. The control groups contained following parameters: no exposure to laser light or TB, treatment only with TB without laser irradiation, or only laser irradiation without previous immersion in TB. Calculations of colony forming units (CFUs) were conducted by using aCOlyte (Synbiosis). Differences in CFUs were analyzed by the Wilcoxon test. In all study groups, the reduction in CFUs was statistically significant. The differences in CFUs before and after intervention were insignificant. The K3 C.a. control group showed a statistical reduction of Candida albicans after laser irradiation. Our study confirmed the efficacy of aPDT against C. albicans, C. glabrata and C. krusei being dependent on the laser parameters and the type of fungus. The advantage of this study is the validation of aPDT effectiveness in in vitro studies to transpose this data into future clinical trials using photodynamic therapy in the treatment of oral candidiasis.

Is a 10-minute surgical scrub necessary in urologic prosthetic surgery? A randomized study of the effect of a 5- vs 10-minute surgical scrub on bacterial colony counts in the genital skin.

To determine the antiseptic efficacy on bacterial colony counts of a 5- vs 10-minute surgical site scrub in urologic surgery. A prospective cohort study was conducted in 101 patients presenting for elective urological procedures. Patients were randomized to a 5- or 10-minute groin scrub with Betadine (povidone-iodine). Skin swabs were taken immediately after skin clipping and following routine painting with Betadine. A third swab was taken after the betadine skin scrub. Bacterial colony counts were reported as a number of colony-forming units (CFUs). The primary outcome measure was a quantitative comparison of CFUs in the two arms. Fifty-three patients were randomized to a 5-minute scrub and 48 to a 10-minute scrub. After Betadine painting, CFUs were present in 38% of patients in the 5-minute group (mean, 33.5 CFU) and in 27% of the 10-minute group (mean, 45.4 CFU). Following the surgical scrub, only 7.5% of the 5-minute group and 8.3% of the 10-minute group had a measurable CFU count of greater than or equal to 1, and colony counts were low in both groups (5- minute group: mean, 1.5 CFU; 10-minute group: mean, 2.0 CFU). There was no significant difference in CFUs following a 5- or 10-minute scrub (P = 0.28). The addition of a surgical skin scrub leads to a fourfold reduction in the skin CFU count compared with Betadine painting. However, there is no difference between the antibacterial effects of a 5- and 10-minute scrub. A 5-minute scrub may be sufficient in urologic prosthetic surgery.

Comparing the antimicrobial efficacy of pediocin with chlorhexidine and calcium hydroxide as intracanal medicaments against persistent root canal infections.

Introduction:The aim of this study is to compare the antimicrobial activity of pediocin with chlorhexidine (CHX) and calcium hydroxide (Ca(OH)2) against Enterococcus faecalis and Staphylococcus epidermidis biofilms.Materials and Methods:The prepared root canals of 80 teeth were contaminated with E. faecalis (n = 40) and S. epidermidis (n = 40) for 21 days to create biofilms. The samples in each group were allocated randomly into the following four subgroups (n = 10) according to the decontamination protocol: Group 1: 1% Pediocin, Group 2: 2% CHX, Group 3: Ca(OH)2, and Group 4: saline (negative control). At 5 days, the antimicrobial efficacy of the medicaments against E. faecalis and S. epidermidis was assessed by collecting dentin shavings from the canal walls created using Gates Glidden drill sizes 4 and 5, corresponding to a depth into the root canal walls of 200 μm and 400 μm, respectively. The total number of colony-forming units (CFUs) was counted. The Wilcoxon signed-rank test was used to compare the difference in CFUs between the two depths (P > 0.05).Results:There was no bacterial growth in samples treated with pediocin, CHX, or Ca(OH)2 at either depth.Conclusion:In this laboratory experimental model, pediocin exhibited the same antimicrobial properties against E. faecalis and S. epidermidis as CHX and Ca(OH)2.

Comparing the effectiveness between gel and foam hand sanitizers


 Background: Hand sanitizers are commonly used as an alternative to washing hands with warm water and soap. There are a variety of different hand sanitizers including gel and foam and they are known to kill several bacteria. Many factors play a role in the effectiveness of hand sanitizers such as the alcohol concentration and the techniques used to apply hand sanitizers. Alcohol based hand sanitizers must have an alcohol concentration of 60 - 70% to be effective. There is currently no legislation regulating hand sanitizers and there is a lack of research focusing on differences between foam and gel hand sanitizers. This research study investigates effectiveness of gel compared to foam hand sanitizers by evaluating the prevalence of Escherichia coli (E.coli) on pigskins. Methods: To compare the hand sanitizers, microbiological sampling was completed. E.coli was introduced onto 65 pigskins. Five pigskins were used as a baseline to determine the average amount of Colony Forming Units (CFUs) of E.coli present prior to the application of hand sanitizers. One set of the 30 pigskins was applied with gel hand sanitizer, whereas the other 30 was applied with foam hand sanitizer. The pigskins were swabbed with QuickSnap swabs and plated onto 3M Petrifilms. The 65 petrifilms were incubated at 35oC for 48 hours. After incubation, the CFUs of E.coli present on the petrifilm were enumerated. The difference in CFUs was calculated to determine the reduction in E.coli and the overall effectiveness of hand sanitizers. Results: The data was analyzed by using the statistical software, NCSS. Statistical analysis showed that the findings were statistically significant and the null hypothesis (Ho: no difference in CFUs of E.coli between foam versus gel alcohol-based hand sanitizers) was rejected with a power of 0.9997 at p-value of 0.00000. This indicates that there is a difference in the ability to reduce E.coli between gel and foam hand sanitizers and gel sanitizers appeared to be more effective. Conclusion: These results indicate that there was a difference in the effectiveness between foam and gel hand sanitizers in reducing E.coli that was inoculated onto pigskins. However, consumers should be aware that hand sanitizers do not completely eliminate all pathogens. Though gel hand sanitizers are more effective, they should only be used when there are no other methods of keeping hands clean.

In vitro antimicrobial efficacy of riboflavin-UVA for common bacteria in ophthalmology

Background Staphylococcus aureus,Staphylococcus epidermidis and Pseudomonas aeruginosa are common pathogenic bacteria in ophthalmology.To seek an optimal antimicrobial method is critical for the prevention of infection.Objective This study was to investigate the antimicrobial efficacy of riboflavin-ultraviolet A (UVA) on Staphylococcus aureus,Staphylococcus epidermidis and Pseudomonas aeruginosa.Methods Ten strains of frozen Staphylococcus aureus,Staphylococcus epidermidis and Pseudomonas aeruginosa were incubated to culture dishes and resuscitated for 2 times.The bacteria were put into bacterial dilution to prepare the suspension of 0.5 Mclntosh concentration and then the suspension was diluted 10-folds.UVA was used to irradiate the culture dishes added 0,0.1％,0.5％ riboflavin for 10,20,30 minutes with the exposure intensities of 1,2,3 mW/cm2,and the optimal bacteriostatic condition was evaluated by measuring the diameters of inhibition zone.Staphylococcus aureus,Staphylococcus epidermidis and Pseudomonas aeruginosa were divided into normal control group,riboflavin group,UVA group and photodynamic therapy (PDT) group (riboflavin+UVA),and colony forming unit was compared among the groups.Results The optional experimental conditions of PDT inactivating microorganisms were determined as UVA wavelength 370 nm,UVA exposure rate 3 mW/cn2,UVA exposure time 30 minutes and riboflavin 0.1％.The colony forming units were decreased in turn in the normal control group,riboflavin group,UVA group and PDT group (F =110.990,190.841,118.197,all at P =0.000).Compared with the normal control group,the colony forming units were significantly decreased in the UVA group (t =14.640,7.550,5.320,all at P＜0.05),and the colony forming units were significantly decreased in the PDT group (t =25.480,14.750,12.920,all at P＜0.05).The colony forming units decreased by 46.4％-50.2％ in the UVA group and 91.6％-96.7％ in the PDT group.There was no significant difference in colony forming units between the normal control group and the riboflavin group(t =0.660,0.470,0.510,all at P＞0.05).Conclusions PDT can effectively inactivate common bacteria in ophthalmology.This result suggests that PDT has a potential in the treatment of microbial keratitis. Key words: Ultraviolet; Riboflavin; Photodynamic therapy; Bacterium; Inactivating microorganism

Efficacy of Atmospheric Pressure Plasma as an Antibacterial Agent Against Enterococcus Faecalis in Vitro

Enterococcus faecalis (E. faecalis) is a microorganism that can survive extreme challenges in obturated root canals. The aim of this study was to evaluate the efficacy of a non-thermal atmospheric pressure plasma plume against E. faecalis in vitro. A non-thermal atmospheric pressure plasma jet device which could generate a cold plasma plume carrying a peak current of 300 mA was used. The antibacterial efficacy of this device against E. faecalis and its biofilm under different conditions was detected. The antibacterial efficacy of the plasma against E. faecalis and Staphylococcus aureus (S. aureus) was also evaluated. After plasma treatment, the average diameter of inhibition zone on S. aureus and E. faecalis was 2.62±0.26 cm and 1.06±0.30 cm, respectively (P < 0.05). The diameter was increased with prolongation of the treatment duration. The diameters of inhibition zone of the sealed Petri dishes were larger than those of the uncovered Petri dishes. There was significant difference in colony-forming units between plasma group and control group on E. faecalis biofilm (P < 0.01). The transmission electron microscopy revealed that the ultrastructural changes cytoderm of E. faecalis were observed after treatment for 2 min. It is concluded that the non-thermal atmospheric pressure plasma could serve as an effective adjunct to standard endodontic microbial treatment.

Analysis of contamination of endodontic absorbent paper points

Purpose: The aim of this study was to assess the contamination status of endodontic absorbent paper points from sterilized or not sterilized commercial packs, as well as paper points exposed to the dental office environment. Methods: Twenty absorbent paper points were evaluated for contamination status packed under different conditions: commercial/sterilized pack, commercial/non-sterilized pack, exposed to the clinical environment, and intentionally contaminated (positive control). Contamination was determined qualitatively and quantitatively by aerobiosis, capnophilic growth, and pour plate. The Petri dishes were analyzed with a colony counter, and the results were expressed as colony-forming units. The data were analyzed by Kruskal-Wallis test (α=0.05). Results: No difference in colony-forming units was found among the groups of endodontic absorbent paper points. All groups were contaminated by fungi and bacteria. Conclusion: It can be concluded that the sterilization of absorbent endodontic paper points before clinical use should be recommended regardless of commercial presentation.

Antimicrobial Efficacy of Two Irrigation Techniques in Tapered and Nontapered Canal Preparations: An In Vitro Study

The aim of this in vitro study was to determine whether irrigation with apical negative pressure was more effective than traditional positive-pressure irrigation in eradicating Enterococcus faecalis from preshaped root canals. Fifty-four extracted mandibular molars were instrumented to produce either a non-tapered or tapered preparation, sterilized, inoculated with E. faecalis for 30 days, and then randomly assigned into the following groups: group 1—non-tapered preparation and negative-pressure irrigation, group 2—non-tapered preparation and positive-pressure irrigation, group 3—tapered preparation and positive-pressure irrigation, and group 4—tapered preparation and negative-pressure irrigation. Mesial canals were sampled before and after final irrigation and samples incubated aerobically for 48 hours at 37°C. Scanning electron microscopic analysis confirmed dense bacterial colonies in the positive control, consistent with biofilm formation. A statistically significant difference was evident when comparing apical negative-pressure irrigation to positive-pressure irrigation (p=0.004). There was no statistically significant difference in colony-forming units (CFUs) between sizes #35 and #45, nor between tapered and non-tapered preparation. The results of this in vitro study showed that apical negative-pressure irrigation has the potential to achieve better microbial control than traditional irrigation delivery systems.

Wedding Rings are not a Significant Source of Bacterial Contamination Following Surgical Scrubbing

Despite some evidence that the wearing of rings may increase the microbial load, there is currently nothing to suggest that viable bacteria remain following a standard surgical scrub. The aim of the study was to examine the distribution and type of microbial flora seen on the hands of doctors following a standard surgical scrub. Ten surgeons and 10 anaesthetists, all of whom wore wedding rings on the fourth finger of their left hand, participated in the study. Each individual was asked to 'scrub-up' as for their normal first scrub of the day. Following completion of washing, the wedding ring was removed, its internal circumference swabbed and the swab placed in a culture medium. Volunteers placed each hand palm-down on separate agar plates. The plates were incubated and the number of colonies counted and classified. The culture plates of one of the anaesthetists were damaged in transit leaving a total of 19 subjects for analysis. In all the palm imprint plates, coagulase-negative staphylococci were grown. One surgeon grew coagulase-negative staphylococci from the ring swab. A Candida spp. from the right hand of one surgeon was grown. There was no statistically significant difference between the number of colony-forming units (CFUs) cultured from the right and left (ring-wearing) hands of the surgeons (P = 0.260) and anaesthetists ( P = 0.345). There was no statistical difference in CFUs when surgeons were compared with anaesthetists (P = 0.383 for right hand and P = 0.234 for left). This preliminary study would suggest that a traditional band wedding ring is not a source of a bacterial load following a standard surgical scrub procedure and, as such, there is no requirement for their removal pre-operatively.

Comparison of Probes for Microbial Contamination Following Use in Periodontal Pockets of Various Depths

It has been speculated that periodontal probes can transmit periodontal pathogens from site to site. The purpose of this study was to evaluate the potential for bacterial transmission as a function of periodontal probe design. Four different periodontal probes were used to measure probing depths ranging from 0 to 3 mm and > or = 4 mm. Following measurement of each pocket, probes were transported to a laboratory and cultured on blood agar, and colony forming units (CFUs) were determined for total aerobic, anaerobic, and dark-pigmented colonies (DPCs). Eight randomly selected probe tips, representing each probe type and probing depth category, were selected for examination by scanning electron microscopy (SEM) to determine distribution and morphotypes of adhering microbes. Results showed no statistically significant main effect for probes with respect to CFUs. However, there was a statistically significant increase in CFUs for deeper pockets when compared to pockets 0 to 3 mm. SEM observations were consistent among groups, regardless of probe design or probing depth. All probes exhibited a rough surface texture. Microbes were observed as single organisms or in aggregates and were adherent along the entire length of the probe up to the maximum probing depth. There was no difference in CFUs obtained as a function of periodontal probes. The rough surfaces of the probes used in this study likely promoted bacterial adherence through two different mechanisms: through scraping of the pocket walls by the roughened surfaces of the probe and by the high surface-free energy of the metal probes, facilitating bacterial adherence.

Pharmacodynamics of amoxicillin/clavulanic acid against Haemophilus influenzae in an in vitro kinetic model: a comparison of different dosage regimens including a pharmacokinetically enhanced formulation

Objective To study the pharmacodynamics of amoxicillin/clavulanic acid against different strains of Haemophilus influenzae in an in vitro kinetic model. The concentrations used corresponded to human serum levels obtained after 875 mg amoxicillin/clavulanic acid given b.i.d, 500/125 mg amoxicillin/clavulanic acid given t.i.d. and those obtained with a pharmacokinetically enhanced formulation containing 1125/125 mg amoxicillin/clavulanic acid (immediate release) and 875 mg amoxicillin (sustained release) given b.i.d.Methods Bacteria at an initial inoculum of 106 colony-forming units (CFU)/mL were exposed to amoxicillin/clavulanic acid with an initial concentration of approximately 15/3 mg/L, 8/3 mg/L simulating the peak levels in humans achieved after a dose of 875/125 mg and 500/125 mg with a half-life of 1 h. In addition, experiments with a 2000/125 mg pharmacokinetically enhanced formulation of amoxicillin/clavulanic acid given b.i.d were performed. A repeated dose was given at 12 h after the initial dose of 875/125 mg and the pharmacokinetically enhanced formulation or at 8 and 16 h after the dose of 500/125 mg. The experiments were performed in an in vitro kinetic model, which consists of a spinner flask with a filter membrane fitted in between the upper part and the bottom part in order to prevent bacterial dilution. The medium is removed from the culture flask, through the filter, at a constant rate with a pump. Repeated samples were taken at intervals of 1–2 h up to 24 h during the experiments for viable counting. One of the strains of H. influenzae was also exposed to a constant concentration corresponding to the peak serum levels obtained after a dose of 500/125 mg.Results The concentrations of amoxicillin in the in vitro kinetic model were as expected. At the end of the experiment (24 h), there was a tendency for a greater bactericidal effect with 500/125 mg t.i.d, as compared to 875/125 b.i.d., with differences in CFUs between the two dosing regimens of 2.6 log10 CFU for H. influenzae LH 2803 and 1.8 log10 CFU for the other clinical strains. However, these differences did not reach statistical significance (P = 0.075 and 0.10, respectively). A statistically significant higher bactericidal effect was seen in the experiments with the pharmacokinetically enhanced formulation in comparison with the b.i.d. regimen both at 8, 16 and 24 h and at 8 and 16 h with the t.i.d. regimen. With the new formulation, no regrowth was seen at 24 h, similar to the results obtained with a constant concentration.Conclusions Neither of the standard dosing regimens of amoxicillin (875/125 mg b.i.d. or 500/125 mg) used in our study, in which the time that the free (non-protein-bound) concentration the MIC (T > MIC) exceeding was less than 50%, was sufficient to achieve a complete bactericidal effect during the first 24 h of treatment. However, a statistically significant difference in bactericidal activity was seen at 8, 16 and 24 h vs. the b.i.d. regimen and at 8 and 16 h vs. the t.i.d. regimen with the pharmacokinetically enhanced formulation. This formulation gave a longer T > MIC (73–79%) of amoxicillin even though the concentration of clavulanic acid was only detectable for 45% of the dosing interval, and complete killing of all strains was obtained after 24 h.

Effect of NaF-, SnF2-, and chlorhexidine-impregnated birch toothpicks on mutans streptococci and pH in approximal dental plaque

The antimicrobial effect of birch toothpicks impregnated with 4% NaF, 8% SnF2, or 2% chlorhexidine was studied both in vitro and in vivo. A non-impregnated toothpick served as a control. In vitro, suspensions of Streptococcus mutans were exposed to the various toothpicks for 20 min and then cultured on blood agar. The results of this susceptibility test revealed the following ranking order with respect to inhibition: chlorhexidine > SnF2 > NaF and non-impregnated; with significant differences in colony-forming units (CFU) between these three groups. In vivo, 12 individuals used the 4 types of toothpick 3 times a day for 5 days in a procedure with a crossover design. Saliva and approximal plaque samples were collected at baseline and on various occasions up to 23 days after the treatment. At the same time, plaque-pH was measured at approximal sites 10 min after rinsing with 10% sucrose. The results of these in vivo experiments revealed lower proportions of mutans streptococci after using all four types of toothpick, but the reduction was significant only after 2 days for the toothpicks impregnated with SnF2 and chlorhexidine (P < 0.05). On the sampling occasions 9 and 23 days after the treatment, the mutans streptococci were more or less back to baseline levels again. In saliva no significant differences in the number of mutans streptococci were found either within or between the four treatments. No significant differences were found regarding decline in the plaque-pH between the NaF-, SnF2-, chlorhexidine-, and non-impregnated toothpicks on any of the sampling occasions.

Quantitative cultures of Candida from mouthwash fluid in HIV-infected patients: a longitudinal study.

The density of Candida colonization in mouthwash fluid of 59 HIV-seropositive patients and 21 controls was determined. No significant difference in colony-forming units was found. No correlation was found between the colonizing density of Candida albicans and the CD4 count among the patients. Twenty-seven of the HIV-seropositive patients were followed for almost 3 years. No difference was found between the number of Candida albicans colony-forming units at the first and second time of sampling. Vaccination with HIV IIIB GP 160 vaccine did not have any influence on the prevalence of Candida albicans.