Salicylate inhibition of acanthamoebal attachment to contact lenses.

PURPOSE: Sodium salicylate has shown potential as a component of contact lens care solutions designed to reduce Acanthamoebal attachment to contact lenses. This study determined the minimum effective concentration required to significantly reduce amoebal attachment. METHODS: Different concentrations of sodium salicylate (10, 15, and 20 mM) were applied during exposure of unworn or bacterial biofilm-coated hydrogel contact lenses to Acanthamoeba castellanii trophozoites. Salicylate was applied at stage 1 intervention during biofilm formation on lenses, at stage 2 intervention during amoebal exposure, or at both stages. RESULTS: A significant reduction in amoebal attachment was achieved when 10 mM salicylate was included during stage 1 alone; however, 15 mM was required for stage 2 intervention to significantly reduce attachment to clean or biofilm-coated lenses. For stages 1 and 2 combined intervention, 10 mM sodium salicylate produced a significant reduction in amoebal attachment. CONCLUSIONS: In situ, within a contact lens case, biofilm formation and amoebal attachment would occur concurrently; therefore, stages 1 and 2 intervention would be closest to the real-life situation, thus indicating that 10 mM of salicylate would be an effective minimum concentration for reducing amoebal attachment to hydrogel contact lenses. Inclusion of components in contact lens care solution, such as sodium salicylate, which reduce Acanthamoebal attachment, has the potential to enhance effectiveness, particularly where amoebicidal efficacy may be limited, thus reducing the risk of contact lens-associated Acanthamoebal infection.

ESCRS study of prophylaxis of postoperative endophthalmitis after cataract surgery: Case for a European multicenter study.

PURPOSE: To present the development and design of the European Society of Cataract & Refractive Surgeons multicenter study of the prevention of postsurgical infective endophthalmitis after phacoemulsification and to describe the process for its successful implementation and conduct. SETTING: Twenty-four ophthalmology units and eye clinics in Austria, Belgium, Germany, Italy, Poland, Portugal, Spain, Turkey, and the United Kingdom, with an administrative office in Ireland, coordinating center in England, and data management and statistical unit in Scotland. METHODS: This partially masked randomized placebo-controlled multinational clinical study was designed to evaluate prospectively the prophylactic effect of intracameral cefuroxime and/or perioperative topical levofloxacin on postoperative endophthalmitis after cataract surgery. Random allocation was based on a 2 x 2 factorial design that included participating centers as a class variable. Real-time electronic data collection monitored study progress and provided weekly outcome tables, monthly recruitment summaries, and quarterly analytical reports for the study's Data Monitoring Committee, which evaluated the safety and efficacy by Internet-based conferences. RESULTS: A 2-year lead time was required to meet harmonized standards of clinical research in the European Union, obtain ministerial authorization in 3 countries, gain institutional approvals at 24 hospitals, and procure indemnity insurance for surgical centers. Informed consent instruments, designed to comply with national health policies, were translated into 8 languages. The use of information technology to collect study data enabled the organizers to evaluate individual eligibility at enrollment, adherence with study medications during and after surgery, and postoperative status during follow-up. CONCLUSION: This international cooperative study provided the opportunity to estimate the current incidence of endophthalmitis after cataract surgery in Europe and determine whether 1 or both of 2 antimicrobial regimens reduces the risk for postsurgical intraocular infection.

ESCRS study of prophylaxis of postoperative endophthalmitis after cataract surgery: Preliminary report of principal results from a European multicenter study.

PURPOSE: To report results in the European Society of Cataract & Refractive Surgeons (ESCRS) multicenter study of the prophylaxis of endophthalmitis after cataract surgery. SETTING: Twenty-four ophthalmology units and eye clinics in Austria, Belgium, Germany, Italy, Poland, Portugal, Spain, Turkey, and the United Kingdom, with an administrative office in Ireland, coordinating center in England, and data management and statistical unit in Scotland. METHODS: This partially masked randomized placebo-controlled multinational clinical study to evaluate prospectively the prophylactic effect of intracameral cefuroxime injection and/or perioperative levofloxacin eyedrops on the incidence of endophthalmitis after phacoemulsification cataract surgery began in September 2003 and was terminated early in January 2006. The study used random allocation of patients in a 2 x 2 factorial design. RESULTS: By the end of 2005, complete follow-up records had been received for 13 698 study patients. Such a clear beneficial effect from the use of intracameral cefuroxime had been observed that it was agreed it would be unethical to continue the study and to wait for the completion of all follow-up procedures before reporting this important result. If total reported cases of endophthalmitis are considered, the incidence rate observed in those treatment groups not receiving cefuroxime prophylaxis (23 cases in 6862 patients) was almost 5 times as high (odds ratio [OR], 4.59; 95% confidence interval [CI], 1.74-12.08; P = .002) as that in the groups receiving this treatment (5 cases in 6836 patients). If only cases proved to be due to infection are considered, the rate was more than 5 times as high (OR, 5.32; 95% CI, 1.55-18.26; P = .008) in the treatment groups not receiving cefuroxime. Although the use of perioperative levofloxacin eyedrops as prophylaxis was also associated with a reduction in the observed incidence rate of postoperative endophthalmitis, this effect was smaller and was not statistically significant, whether total reported cases or only cases proven to be due to infection are used in calculating the rates. As not all follow-up procedures are complete, it is possible that further cases of endophthalmitis may be reported; however, it is not expected that this will alter the main conclusion. Nevertheless, it is anticipated that successful completion of follow-up procedures in all patients will increase the total number in the study to approximately 16,000. CONCLUSION: Intracameral cefuroxime administered at the time of surgery significantly reduced the risk for developing endophthalmitis after cataract surgery.

Molecular and physiological evaluation of subtropical environmental isolates of Acanthamoeba spp., causal agent of Acanthamoeba keratitis.

Previous molecular examination of Acanthamoeba spp. has resulted in the determination of distinct genotypes in this genus (designated T1-T12, T14). Genotype T4 has been responsible for the majority of cases of Acanthamoeba keratitis. Here we examine the relative abundance of environmental T4 isolates on beaches and ask whether they have temperature and salinity tolerances that could enhance pathogenicity. Twenty-four Acanthamoeba strains were isolated from beach sand (n = 20), soil (n = 3), and tap water (n = 1) in south Florida. Phylogenetic analysis identified 19 of 24 isolates as T4, the Acanthamoeba keratitis-associated genotype. The remaining isolates were genotype T5 (4) and T11 (1). Nearly all beach isolates were genotype T4, whereas the tap water and soil isolates were mostly T5. All amoebae grew at 0, 1.0, and 2.0% salt and 19 of 20 beach isolates also grew at 3.2%. No soil or tap-water acanthamoebae reproduced at 3.2%. All isolates grew at 37 degrees C and two (T5) at 42 degrees C. Little correlation existed between beach location, salt-tolerance, and genetic relatedness. Overall, the large majority of environmental isolates obtained were genotype T4, suggesting it may be the most common genotype in this environment and could be a potential source of Acanthamoeba keratitis infections.

Determination of amoebicidal activities of multipurpose contact lens solutions by using a most probable number enumeration technique.

Six multipurpose contact lens solutions [All-in-One, All-in-One (Light), ReNu MultiPlus, Optifree Express, Complete, and Solo-care soft] were tested for their efficacies against Acanthamoeba castellanii trophozoites and cysts by using a most probable number (MPN) technique for amoebic enumeration. Against trophozoites, All-in-One, ReNu Multiplus, and Optifree Express achieved total kill (log reduction of >3) after the manufacturer's minimum recommended disinfection time (MMRDT), with the remaining solutions failing to reach a log reduction of 1. After 24 h of exposure, all solutions proved trophozoiticidal, achieving, with the exception of Complete (log reduction of 3.13), total kill. Against cysts, All-in-One gave a log reduction of >3 within the MMRDT, with all other solutions failing to achieve a log reduction of 1. After 24 h of exposure, All-in-One achieved total kill of cysts (log reduction of 3.74), ReNu MultiPlus gave a log reduction of 3.15, and the remaining solutions reached log reductions of between 1.09 and 2.27. The MPN technique provides a simple, reliable, and reproducible method of amoebic enumeration that depends on simply establishing the presence or absence of growth on culture plates inoculated with a series of dilutions and determining the MPN of amoebae present from statistical tables. By use of this technique, two of the multipurpose solutions tested, ReNu MultiPlus and Optifree Express, demonstrated effective trophozoiticidal activities within the recommended disinfection times; however, only All-in-One proved effective against both trophozoites and cysts over the same time period. This MPN technique, which uses axenically produced trophozoites and mature, double-walled cysts, has the potential to form the basis of a national standard for amoebicidal efficacy testing of multipurpose contact lens disinfecting solutions.

Surface treatment or material characteristic: the reason for the high level of Acanthamoeba attachment to silicone hydrogel contact lenses.

PURPOSE: To determine the reason for the high level of attachment of Acanthamebic to silicone hydrogel (SH) contact lenses. The effect surface treatment has on attachment is determined using silicone elastomer (SE) lenses. METHODS: All test lenses were unworn. SH (PureVision), conventional hydrogel (Acuvue), treated SE (Silsoft), and untreated SE (Silsoft) lens quarters were incubated for 90 min with plate-cultured Acanthamoeba castellanii trophozoites. After incubation and rinsing, the trophozoites attached to one surface of each quarter were counted by direct light microscopy. Sixteen replicates were performed for each lens type. Logarithmic transformation of data allowed the use of parametric analysis of variance. RESULTS: Lens polymer had a significant effect on attachment (P<0.001), with higher numbers of trophozoites attaching to the SH and SE (treated and untreated) lenses as compared with the conventional hydrogel. No significant difference in attachment was detected between the SH and SE (treated and untreated) lenses. CONCLUSION: Acanthamoeba attachment to the SH lens was significantly greater than to the conventional hydrogel. The similarity in attachment to surface-treated and non-surface-treated SE lenses suggests that the increased attachment found with the SH lens may be an inherent characteristic of the polymer rather than an effect of the surface treatment procedure. It is possible that SH lenses are at greater risk of promoting Acanthamoeba infection if exposed to the organism because of the enhanced attachment characteristic of this new material.

Enhanced attachment of acanthamoeba to extended-wear silicone hydrogel contact lenses: a new risk factor for infection?

PURPOSE: To establish if silicone hydrogel (S-H) contact lenses could be a risk factor for Acanthamoeba infection by facilitating the attachment of trophozoites to their surface and transfer to the cornea and to determine the effect Acanthamoeba culture technique, patient wear, and Pseudomonas biofilm coating have on attachment to the S-H lens. DESIGN: Experimental material study. PARTICIPANTS AND CONTROLS: Attachment to a S-H lens was compared with that of a conventional hydrogel control lens. Sixteen replicates were carried out for both lens types under each test condition. METHODS: Unworn S-H (PureVision; Bausch & Lomb, Kingston-Upon-Thames UK) and conventional hydrogel (Acuvue; Vistakon, Johnson & Johnson, Jacksonville, FL USA) lens quarters were incubated for 90 minutes in suspensions of liquid or plate-cultured Acanthamoeba castellanii trophozoites. Unworn, worn, and Pseudomonas biofilm coated S-H and hydrogel quarters were incubated for 90 minutes with plate-cultured trophozoites. MAIN OUTCOME MEASURES: Trophozoites attached to one surface of each lens quarter were counted by direct light microscopy. Logarithmic transformation of data allowed the use of a parametric analysis of variance. RESULTS: Lens polymer had a significant effect on attachment (P < 0.001), with higher numbers of trophozoites attaching to the S-H lens. Culture technique also had a significant effect on attachment (P = 0.013), with higher numbers of liquid-cultured organisms attaching to both lens types. A significant increase in attachment was demonstrated with worn and Pseudomonas biofilm-coated hydrogel lenses (P < 0.001); however, this difference was not seen with the S-H lens. CONCLUSIONS: Acanthamoebal attachment to the S-H lenses was significantly greater than to the conventional hydrogel. Liquid-cultured trophozoites demonstrated a higher affinity for the lenses tested. Wear and bacterial biofilm coating had no effect on attachment to S-H lenses. The increased attachment found with the S-H lens may be an inherent characteristic of the polymer or a side effect of the surface treatment procedure to which the lenses are exposed. It is possible that S-H lenses are at greater risk of promoting Acanthamoeba infection if exposed to the organism because of the enhanced attachment characteristic of this new material.