Adhesion of Acanthamoeba on Scleral Contact Lenses According to Lens Shape.

Purpose: To investigate the adhesion of Acanthamoeba to scleral contact lens (ScCL) surface according to lens shape. Methods: Two strains of A. polyphaga (CDC:V062 and ATCC 30461) and one clinical Acanthamoeba isolate, were inoculated onto five contact lens (CL): one first-generation silicone hydrogel (SHCL; lotrafilcon B; adhesion control) containing plasma surface treatment; two ScCL (fluorosilicone acrylate) one containing surface treatment composed of plasma and the other containing plasma with Hydra-PEG, and two CL designed with a flat shape having the same material and surface treatments of the ScCL. Trophozoites that adhered to the lens's surfaces were counted by inverted optical light microscopy. Possible alterations of the lens surface that could predispose amoeba adhesion and Acanthamoeba attached to these lens surfaces were evaluated by scanning electron microscopy (SEM). Results: All strains revealed greater adhesion to the ScCL when compared with the flat lenses (P < 0.001). The clinical isolate and the ATCC 30461 had a higher adhesion (P < 0.001) when compared with the CDC:V062. A rough texture was observed on the surface of the lenses that have been examined by SEM. Also, SEM revealed that the isolates had a rounded appearance on the surface of the ScCL in contrast with an elongated appearance on the surface of the silicone hydrogel. Conclusions: The findings revealed that the curved shape of the ScCL favors amoeba adhesion.

Risk Factors for Acanthamoeba Keratitis-A Multistate Case-Control Study, 2008-2011.

OBJECTIVE: To identify modifiable risk factors contributing to Acanthamoeba keratitis (AK) infection. METHODS: A case-control investigation was conducted. Case patients were soft contact lens wearers with laboratory-confirmed AK. Control were soft contact lens wearers ≥12 years of age, with no history of AK. Case patients were recruited from 14 ophthalmology referral centers and a clinical laboratory. Control were matched on state of residence and type of primary eye care provider (ophthalmologist or optometrist). Participants were interviewed using a standardized questionnaire. Univariable and multivariable conditional logistic regression analyses were conducted. Matched odds ratios (mORs) were calculated. RESULTS: Participants included 88 case patients and 151 matched control. Case patients were more likely to be aged <25 years (unadjusted mOR 2.7, 95% confidence interval 1.3-5.5) or aged >53 years (mOR 2.5, 1.1-5.7), and more likely to be men (mOR 2.6, 1.4-4.8). Unadjusted analyses identified multiple risk factors: rinsing (mOR 6.3, 1.3-29.9) and storing lenses in tap water (mOR 3.9, 1.2-12.3), topping off solution in the lens case (mOR 4.0, 2.0-8.0), having worn lenses ≤5 years (mOR 2.4, 1.3-4.4), rinsing the case with tap water before storing lenses (mOR 2.1, 1.1-4.1), and using hydrogen peroxide (mOR 3.6, 1.1-11.7) versus multipurpose solution. Significant risk factors in multivariable modeling included age >53 years, male sex, topping off, and using saline solution. CONCLUSIONS: Numerous modifiable risk factors for AK were identified, mostly involving hygiene practices. To reduce the risk of AK, lens wearers should observe recommended lens care practices.

Acanthamoeba keratitis in noncompliant soft contact lenses users: Genotyping and risk factors, a study from Cairo, Egypt.

Acanthamoeba keratitis (AK) is a severe corneal infection that may occur as a serious outcome of improper use of contact lenses (CL). OBJECTIVES: The study aimed to diagnose AK in soft CL users presenting with infectious keratitis, and to identify the prevalent genotypes isolated from different cases. Another aim was to determine the CL hygiene-related risk behaviors, and to explore the risk of water exposure for developing AK. METHODS: A cross sectional study was performed. A questionnaire was carried out including 260 clinically diagnosed cases as infectious keratitis (170 females and 90 males); all of them were soft CL users for the suspected risk factors. Corneal scrapes from the affected eyes were cultured to diagnose bacterial and AK. PCR was performed and the amplified products were sequenced and compared with GenBank data. RESULTS: The parasite was positively amplified from 32 samples (12.3%). Acanthamoeba T4 genotype was identified in 27/32 (84.4%) of isolates. Other detected genotypes belonged to T5 and T3 genotypes at rates of 9.4%, and 6.25%, respectively. The most important risk factors associated with development of AK were female sex, sleeping while wearing CL, and exposure to water resources through different practices. These practices included rinsing the CL case in tap water, swimming and/or showering while wearing CL, using multipurpose solution for cleaning the lenses, using water from over-building tanks. Rubbing the eyes due to discomfort when applying CL was an additional important risk factor associated with AK. The protective factor was regular hand washing before using CL. CONCLUSION: CL users were more exposed to AK and should gain enough health education regarding proper lens hygiene and dangers of tap water exposure.

Failure of molecular diagnostics of a keratitis-inducing Acanthamoeba strain.

An otherwise healthy 49-year-old female patient presented at the local hospital with severe keratitis in both inflamed eyes. She was a contact lens wearer and had no history of a corneal trauma. In our laboratory for medical parasitology Acanthamoebae were detected microscopically from the cornea scraping and from the fluid of the contact lens storage case after xenical culture and showed the typical cyst morphology of Acanthamoebae group II. The diagnosis of "Acanthamoeba keratitis" was established and successful therapy was provided. While the morphological microscopic method led to the correct diagnosis in this case, an in-house multiplex qPCR and a commercial qPCR showed false negative results regarding Acanthamoeba sp. The subsequent sequencing revealed the Acanthamoeba genotype T4. In the present case report, the inability to detect Acanthamoebae using qPCR only is presented. Therefore, we recommend the utilization of combined different assays for optimal diagnostic purposes.

Effect of Multipurpose Solution Combined With Autophagy Inhibitors on Adhesion of Acanthamoeba trophozoites to Silicone Hydrogel Contact Lenses.

PURPOSE: The aim of the study was to evaluate the effect of multipurpose contact lens (CL) solution (MPS) combined with autophagy inhibitors on the adhesion properties of Acanthamoeba castellanii (AC) trophozoites to silicone hydrogel CLs. METHODS: AC trophozoites were inoculated onto discs trimmed from a silicone hydrogel CL with plasma surface treatment. After 18 hours, the number of adherent AC trophozoites on the CL treated with MPS was counted under phase-contrast microscopy. We analyzed the efficacy of MPS combined with autophagy inhibitors, 3-methyladenine (0.1, 1, and 5 mM) and chloroquine (10, 100 µM, and 1 mM), on Acanthamoeba adhesion by electron microscopy. RESULTS: Adhesion of AC trophozoites to the CL treated with MPS (average number of trophozoites adhered to the CL: 61.2 ± 8.1) was significantly lower compared with that of the CL treated without MPS (83.8 ± 10.2) (P = 0.027). In MPS application, the number of adhered AC trophozoites treated with 3-methyladenine [8.2 ± 2.5 for 5 mM (P = 0.008)] or chloroquine [19.4 ± 7.6 for 100 µM (P = 0.038) and 5.3 ± 1.9 for 1 mM (P = 0.001)] was significantly reduced compared with the sample without autophagy inhibitors. However, the number of adherent AC trophozoites was not significantly reduced in less than 5 mM in 3-methyladenine [50.4 ± 5.1 for 0.1 mM (P = 0.084) and 43.1 ± 5.0 for 1 mM (P = 0.079)] and 100 µM in chloroquine [40.6 ± 13.5 for 10 µM (P = 0.075)]. 3-methyladenine induced blebby structures or disrupted the membranes of AC trophozoites. AC trophozoites treated with chloroquine showed undigested organelles in the cytoplasm of Acanthamoeba cells. CONCLUSIONS: MPS combined with 3-methyladenine or chloroquine reduced the adhesion rate of AC trophozoites rather than MPS containing only polyhexamethylene biguanide. Appropriate concentrations of autophagy inhibitors, 3-methyladenine and chloroquine, added to commercial MPS should be considered to decrease the clinical rate of Acanthamoeba keratitis.

Water Exposure is a Common Risk Behavior Among Soft and Gas-Permeable Contact Lens Wearers.

PURPOSE: To understand soft contact lens (SCL) and gas-permeable (GP) lens wearers' behaviors and knowledge regarding exposure of lenses to water. METHODS: The Contact Lens Risk Survey (CLRS) and health behavior questions were completed online by a convenience sample of 1056 SCL and 85 GP lens wearers aged 20 to 76 years. Participants were asked about exposing their lenses to water and their understanding of risks associated with these behaviors. Chi-square analyses examined relationships between patient behaviors and perceptions. RESULTS: GP lens wearers were more likely than SCL wearers to ever rinse or store lenses in water (rinsing: 91% GP, 31% SCL, P < 0.001; storing: 33% GP, 15% SCL P < 0.001). Among SCL wearers, men were more likely to store (24% vs. 13%, P = 0.003) or rinse (41% vs. 29%, P = 0.012) their lenses in water. Showering while wearing lenses was more common in SCL wearers (86%) than GP lens wearers (67%) (P < 0.0001). Swimming while wearing lenses was reported by 62% of SCL wearers and 48% of GP lens wearers (P = 0.027). Wearers who rinsed (SCL; P < 0.0001, GP; P = 0.11) or stored lenses in water (SCL; P < 0.0001, GP P = 0.007) reported that this behavior had little or no effect on their infection risk, compared with those who did not. Both SCL (P < 0.0001) and GP lens wearers (P < 0.0001) perceived that distilled water was safer than tap water for storing or rinsing lenses. CONCLUSIONS: Despite previously published evidence of Acanthamoeba keratitis' association with water exposure, most SCL, and nearly all GP lens wearers, regularly expose their lenses to water, with many unaware of the risk.

A Rabbit Model of Acanthamoeba Keratitis: Use of Infected Soft Contact Lenses After Corneal Epithelium Debridement With a Diamond Burr.

Purpose: To develop a rabbit model of Acanthamoeba keratitis (AK) as the best method to reproduce the natural course of this disease. Methods: To induce AK, infected contact lenses (1000 amoebae/mm2, 90% trophozoites) were placed over the previously debrided corneal surface, in combination with a temporary tarsorrhaphy. Environmental and clinical strains of Acanthamoeba spp. (genotype T4) were used. Three groups (1L, n = 32; 2L-21d, n = 5; 2L-3d, n = 23) were established according to the number of contact lenses used (1L, 1 lens; 2L-21d and 2L-3d, 2 lenses) and the placement day of these (1L, day 1; 2L-21d, days 1 and 21; 2L-3d, days 1 and 3). The infection was quantified by a clinical score system and confirmed using corneal cytology and culture, polymerase chain reaction and histopathologic analysis. Results: The infection rate obtained was high (1L, 87.5%; 2L-21d, 100%; 2L-3d, 82.6%), although no clinical signs were observed in the 50% of the infected animals in group 1L. Among groups, group 2L-3d showed more cases of moderate and severe infection. Among strains, no statistically significant differences were found in the infection rate. In the control eyes, cross infection was confirmed when a sterile contact lens was placed in the previously debrided corneas but not if the eye remained intact. Conclusions: The combination of two infected contact lenses after corneal debridement seems to be an alternative model, clinically and histopathologically similar to its human counterpart, to induce the different AK stages and reproduce the course of the disease in rabbits.

Painless Acanthamoeba Keratitis with Normal Vision.

PURPOSE: To report an atypical case of contact lens-related Acanthamoeba keratitis. CASE REPORT: A 15-year-old secondary school female student with a history of soft contact lens wear was referred to our hospital by a private general practitioner for management of right eye redness and discomfort for 2 weeks. Upon examination, the best-corrected visual acuity was 20/20 and 20/16 for her right and left eyes, respectively. There was diffuse radial keratoneuritis noted in the cornea of her right eye without any associated epithelial defect. Corneal scraping was performed to induce epithelial defect over the keratoneuritis area, and it was positive for Acanthamoeba trophozoites. She was treated with amoebicidal therapy consisting of propamidine isethionate 0.1% and polyhexamethylene biguanide 0.02%, for 6 months. The patient did not complain of any ocular pain in the entire course of her disease. She attained a final visual acuity of 20/13 in the affected eye with residual peripheral radial perineuritic scar. CONCLUSIONS: Atypical presentation of Acanthamoeba infection is uncommon. This case should arouse the awareness of an indolent presentation of this potentially sight-threatening disease. Clinicians should have a high level of suspicion in contact lens users who present with corneal abnormalities despite an absence of pain. Microbiological work-up and prompt treatment led to a complete resolution of Acanthamoeba infection in our patient.

Effects of Lead Phytochemicals of Radix Scutellariae on Acanthamoeba.

Purpose: Acanthamoeba keratitis (AK), which is associated with noncompliant use of contact lenses, remains difficult to treat due to delayed diagnosis and paucity of therapeutic agents. Although improvements in activity against Acanthamoeba infection have been achieved in disinfecting solutions for soft contact lenses, such modifications have not been extended to those for special rigid gas permeable (RGP) contact lenses, which are increasingly used for myopia control in children. Phytochemicals present in herbs used for traditional Chinese medicine may be effective as therapeutic or preventive agents. The purpose of this study was to investigate amoebicidal properties of lead phytochemicals of Radix scutellariae alone and in combination with multipurpose (disinfecting) solutions (MPS) for RGP lenses. Methods: Viability of Acanthamoeba castellani and A. polyphaga trophozoites was determined following exposure to four phytochemicals: baicalin, baicalein, wogonoside, and oroxylin A and both alone and in combination with four RGP MPS, using a modified stand-alone technique. Results: As individual agents, wogonoside and oroxylin A showed highest activity against A. castellani and A. polyphaga trophozoites, respectively. For both organisms, the combination of baicalein and oroxylin A was superior. Effectiveness of MPS alone did not exceed 0.27 log reduction, but addition of combined baicalein and oroxylin A resulted in 0.92 and 0.64 log reductions of A. castellani and A. polyphaga, respectively. Conclusions: The combination of baicalein and oroxylin A enhanced the activity of MPS for RGP contact lenses against trophozoites of two pathogens, A. castellani, and A. polyphaga, and offers a potential therapeutic and/or preventative agent for AK.

Rare Case of Polymicrobial Keratitis With Balantidium coli.

PURPOSE: To report a rare case of polymicrobial keratitis due to Balantidium coli and gram-negative bacteria, Pseudomonas aeruginosa and Klebsiella pneumoniae, in a soft contact lens (CL) wearer. METHODS: We report a case of CL-related keratitis due to B. coli, P. aeruginosa, and K. pneumoniae. RESULTS: The culture of the corneal scrapings, the CL cleaning solution, and the CL revealed the growth of a rare ciliated parasite, B. coli, along with gram-negative bacteria, namely, P. aeruginosa and K. pneumoniae. The patient was successfully treated with topical broad-spectrum antibiotics and intravenous metronidazole. CONCLUSIONS: Polymicrobial keratitis has seldom been reported with B. coli as the causative agent. CL wear can be a risk factor for this infection. Treatment with topical antibiotics may not suffice, and the intravenous route of antiprotozoal drugs may be a useful adjunct. Increasing awareness, early diagnosis, and treatment may improve the final visual outcome.

Adhesion of Acanthamoeba on Silicone Hydrogel Contact Lenses.

PURPOSE: To evaluate adhesion of Acanthamoeba trophozoites to different silicone hydrogel contact lens (SHCL) generations with and without multipurpose contact lens care solution (MPS) treatment. METHODS: Acanthamoeba lugdunensis L3a trophozoites were inoculated onto discs trimmed from SHCLs: first generation, Air Optix (Lotrafilcon B) with a plasma surface treatment, second generation, Acuvue Oasys (Senofilcon A), which contains an internal wetting agent (Hydraclear), and third generation, Biofinity (Comfilcon A) with no surface treatment. After 18-hour inoculation, the number of adherent trophozoites on SHCLs was counted as the control under phase contrast microscopy. The effects of the 3 different MPSs, Opti-Free Express, ReNu Fresh, and Biotrue, soaking SHCLs for 6 hours, on Acanthamoeba adhesion were analyzed. Scanning electron microscopic examination was performed for assessment of Acanthamoeba attached on the lens surface. RESULTS: Acanthamoeba trophozoites showed greater adhesion to Air Optix than to Acuvue Oasys and Biofinity (P < 0.05). On Air Optix and Acuvue Oasys, the number of adherent Acanthamoeba was significantly reduced compared with the control after treatment with Opti-Free Express (P < 0.05), but not significantly reduced by treatment with ReNu Fresh and Biotrue (P > 0.05). Acanthamoeba did not adhere to Biofinity regardless of MPSs treatment. Attachment of the acanthopodia of Acanthamoeba on the curved ridge of the Air Optix lens surface was observed. CONCLUSIONS: Acanthamoeba showed greater affinity for the first-generation SHCL and seemed to be more attached on SHCLs with more ridges. MPS with myristamidopropyl dimethylamine reduced the adhesion rate.

Demodex sp. as a Potential Cause of the Abandonment of Soft Contact Lenses by Their Existing Users.

Demodex mites may be a potential etiological factor in the development of various eye and skin disorders. The aim of the study was to investigate the presence of Demodex in the hair follicles of eyelashes and their potential influence on abandoning soft contact lenses which had been previously well tolerated by their users. A group of 62 users of contact lenses (28 with emerging discomfort and 34 without discomfort) were examined. There is a need to check the existence of a relationship between D. folliculorum or/and D. brevis infestation and the emergence of intolerance to the presence of soft contact lenses. The removed lashes were examined under light microscopy, applying standard parasitological methods if demodicosis is suspected. A positive result was assumed if at least one adult stage, larva, protonymph/nymph, or egg of D. folliculorum and/or D. brevis was present. A positive correlation was observed between the presence of Demodex and intolerance to contact lenses by their existing users (p < 0.05), and Demodex sp. infections were observed in 92.86% of patients with intolerance to contact lenses. Our results provide further evidence for the pathogenic role played by the mites in the development of eye diseases.

In vivo imaging of radial keratoneuritis in patients with Acanthamoeba keratitis by anterior-segment optical coherence tomography.

PURPOSE: To investigate in vivo corneal changes of radial keratoneuritis in early-stage Acanthamoeba keratitis (AK) using anterior-segment optical coherence tomography (AS-OCT). DESIGN: Single-center, prospective clinical study. PARTICIPANTS: Four eyes (4 patients with a mean age of 28.5 years) with early-stage AK showing radial keratoneuritis were included in this study. Definitive diagnosis was made by confirmation of AK cysts using in vivo confocal microscopy and culture. METHODS: Anterior-segment OCT examination was performed on the initial visit and at follow-up visits paying special attention to radial keratoneuritis. MAIN OUTCOME MEASURES: Selected AS-OCT images of the cornea were evaluated qualitatively for the shape and degree of light reflection of abnormal neurons. RESULTS: With the use of AS-OCT, we successfully obtained high-resolution images of putative radial keratoneuritis in all patients as highly reflective bands or lines in the corneal stroma. The depth and width of the highly reflective bands/lines varied from case to case (anterior stroma to mid-stroma, from 20 to 200 µm). Some lines ran obliquely from the deep peripheral stroma toward the anterior stroma, and some were located at different depths (subepithelial and mid-stroma) and ran relatively parallel to the corneal layers. After appropriate treatment, radial keratoneuritis was resolved by both slit-lamp biomicroscopy and AS-OCT in all patients. CONCLUSIONS: High-resolution Fourier-domain AS-OCT provides novel and detailed visual information of radial keratoneuritis in patients with early-stage AK. Visualization of radial keratoneuritis by AS-OCT may be a useful adjunct to the diagnosis and follow-up of early-stage AK.

Failure of chemotherapy in the first reported cases of Acanthamoeba keratitis in Pakistan.

Acanthamoeba keratitis is a painful and progressive infection of the cornea that can result in loss of vision. Here, for the first time in Pakistan, we report two cases of Acanthamoeba keratitis. The first patient was a 37-year-old female who presented with severe itching, redness, pain, along with loss of vision. The patient was a regular soft contact lens wearer. The second patient was a 25-year-old female who had been using soft contact lenses for the past two years. She presented with a burning sensation and extreme pain, along with loss of vision. Both patients were treated for a possible microbial keratitis with topical moxifloxacin hydrochloride drops, vancomycin drops, propamidine isethionate ointment, amphotericin B drops, and amikacin drops. However, the response was inadequate and both patients were referred for corneal transplant. Acanthamoeba castellanii was isolated by placing contact lenses and contact lens cases on non-nutrient agar plates containing a lawn of non-invasive Escherichia coli K-12 HB101 bacteria. The polymerase chain reaction (PCR) using genus-specific probes confirmed the identity of Acanthamoeba spp., whereas the morphological characteristics of trophozoites and cysts were suggestive of A. castellanii in both cases. With growing use of contact lenses for vision correction/cosmetic use coupled with sub-standard lens care in this region and the possibility of non-contact lens-associated Acanthamoeba keratitis, a need for increased awareness of this sight-threatening infection is discussed further.

In vivo laser confocal microscopy findings of radial keratoneuritis in patients with early stage Acanthamoeba keratitis.

OBJECTIVE: To investigate in vivo corneal changes of keratoneuritis in early stage Acanthamoeba keratitis (AK) using in vivo laser confocal microscopy. DESIGN: Single-center, prospective, clinical study. PARTICIPANTS: Thirteen eyes (12 patients; 5 men and 7 women; mean age ± standard deviation, 22.3 ± 4.2 years) with keratoneuritis resulting from early stage AK were included in this study. TESTING: In vivo laser confocal microscopy was performed, paying special attention to keratoneuritis. MAIN OUTCOME MEASURES: Selected confocal images of corneal layers were evaluated qualitatively for shape and degree of light reflection of abnormal cells and deposits. RESULTS: In all patients, Acanthamoeba cysts were observed clearly in the basal epithelial cell layer as highly reflective round particles with a diameter of 10 to 20 µm. Bowman's layer infiltration of Acanthamoeba cysts was observed in only 1 case, and no cases showed stromal or nerve infiltration of Acanthamoeba cysts. In the stroma, all cases showed highly reflective activated keratocytes forming a honeycomb pattern; these changes were significant around the keratoneuritis. Infiltration of inflammatory cells, possibly polymorphonuclear cells, was observed along with keratocyte bodies in all cases. Numerous highly reflective spindle-shaped materials were observed around the keratoneuritis. Most notably, highly reflective patchy lesions were observed around the keratoneuritis in 11 cases (84.6%). Inflammatory cells also were observed in the endothelial cell layer in 4 cases (30.8%). CONCLUSIONS: In vivo laser confocal microscopy identified consistent corneal abnormalities around keratoneuritis in early stage AK patients, of which highly reflective patchy lesions may be characteristic of keratoneuritis. Further morphologic studies of corneas with early stage AK in a larger number of patients may elucidate the clinical significance of radial keratoneuritis and may help us to understand the interaction between Acanthamoeba organisms and host corneal cells or nerves.

Effects of multipurpose contact lens care solutions on the adhesion of Acanthamoeba to silicone hydrogel contact lenses.

PURPOSE: To evaluate the effect of 7 multipurpose contact lens care solutions (MPSs) on the adhesion of Acanthamoeba (AC) to 5 silicone hydrogel contact lenses (SHCLs). METHODS: Acanthamoeba castellanii (ATCC50370) trophozoites were inoculated onto disks trimmed from SHCLs, Asmofilcon A, Galyfilcon A, Senofilcon A, Lotrafilcon B, and Balafilcon A. After 4-hour incubation, the number of adherent AC trophozoites on SHCL was counted under phase contrast microscopy. AC trophozoites mixed with 7 MPSs were inoculated onto Balafilcon A and incubated for 24 hours followed by direct counting, phase contrast microscopy, and scanning electron microscopy. AC cysts were also inoculated onto Balafilcon A followed by counting using phase contrast microscopy. RESULTS: Adhesion of AC trophozoites to Lotrafilcon B and Balafilcon A was 10 times higher in comparison with the other 3 SHCLs. Twenty four-hour treatment of AC trophozoites with Epica Cold, Epica Cold Aquamore, ReNu MultiPlus, OptiFree Plus, and Complete DoubleMoist reduced the numbers of adherent AC to less than 25% of control, whereas the numbers of AC treated with Complete AminoMoist and C3 SoftOne Moist was about 50% and 75% of control, respectively. Normal AC trophozoites without any treatments showed 25 times higher adhesion rates compared with normal AC cysts. CONCLUSIONS: The adhesion rates of AC trophozoites to SHCL varied depending on the type of MPSs used. Appropriate uses of MPS could reduce adhesion rates of AC to SHCL and potentially decrease clinical rates of Acanthamoeba keratitis.

Prognostic factors in Acanthamoeba keratitis.

OBJECTIVE: To assess the prognostic factors influencing visual prognosis and length of treatment after acanthamoeba keratitis (AK). PARTICIPANTS: Forty-two AK eyes of 41 patients treated between 1999 and 2006 were included. METHODS: A diagnosis of AK was made on the basis of culture results with a corresponding clinical presentation. We calculated the prognostic effect of the various factors on final visual acuity and the length of treatment. Multivariate regression analysis was used to adjust for the simultaneous effects of the various prognostic factors. RESULTS: Mean follow-up was 19.7 ± 21.0 months. Sixty-four percent of cases had > 1 identified risk factor for AK, the most common risk factor being contact lens wear (92.9% of eyes). At presentation, median best spectacle corrected visual acuity (BCVA) was 20/200 (20/30 to Hand Motion [HM]) that improved after treatment to 20/50 (20/20 to Counting Fingers [CF]). Infection acquired by swimming or related to contact lenses had significantly better final BCVA (p = 0.03 and p = 0.007, respectively). Neuritis and pseudodendrites were also associated with better final BCVA (p = 0.04 and p = 0.05, respectively). Having had an epithelial defect on presentation and having been treated with topical steroid were associated with worse final best spectacle corrected visual acuity (BSCVA) (p = 0.0006 and p = 0.04). Multivariate regression analysis found a good initial visual acuity (p = 0.002), infections related to swimming (p = 0.01), the absence of an epithelial defect (p = 0.03), having been treated with chlorhexidine (p = 0.05), and not having receive steroids (p = 0.003) to significantly forecast a good final BCVA. CONCLUSIONS: We identified several prognostic factors that can help clinicians evaluate the expected visual damage of the AK infection and thus tailor treatment accordingly.

Characteristics of an Acanthamoeba keratitis outbreak in British Columbia between 2003 and 2007.

OBJECTIVE: Quantify and describe Acanthamoeba keratitis (AK) cases in British Columbia (BC). DESIGN: A comparison of annual incidence rates confirms the presence of an outbreak. A case series describes characteristics of the outbreak. PARTICIPANTS: All laboratory-confirmed AK cases (persons) in BC (1988-2011; n = 68) were included in the incidence rate comparison. Of the 42 cases (persons) between 2003 and 2007, 32 were selected to interview (laboratory confirmed, 2005-2007), and the 23 who completed interviews form the case series. METHODS: A comparison of standardized annual incidence rates in historic to outbreak periods is performed by z-score test. A telephone interview and descriptive analysis detailing demographics, risk factors, and contact lens (CL) wearing habits was completed for 23 cases. MAIN OUTCOME MEASURES: We measure number of laboratory confirmed cases in BC. In addition, risk factors and potential exposures of these cases are reported. RESULTS: The annual incidence of AK increased significantly from 0.029 to 0.200 per 100 000 population between historic years (1988-2002) and outbreak years (2003-2007; P = 0.022). The annual incidence of AK has since returned to near historic levels (0.056/100 000 population). The case series identified multiple risk factors, including the use of a specific recalled solution (60.9%), daily soft CL wear (95.7%), all-in-one solutions (95.7%), showering while wearing CL (65.2%), and generally poor CL hygiene. CONCLUSIONS: A significant increase in annual AK incidence occurred between 2003 and 2007 in BC. After 2007, the incidence of AK returned to near historic levels. The recalled solution was associated with many cases; however, other risk factors were also identified, including being unaware of the recall and poor CL hygiene practices, highlighting the need for improved education about the severity of AK and consequences of improper CL hygiene. FINANCIAL DISCLOSURE(S): The authors have no proprietary or commercial interest in any of the materials discussed in this article.

Amoebicidal effects of contact lens disinfecting solutions.

PURPOSE: To compare the traditional manual hemacytometer method and an automated counter (Vi-cell) to enumerate and distinguish between viable and non-viable amoeba, and to determine the efficacies of contact lens (CL) disinfecting solutions against three species of Acanthamoeba. The efficacies in the presence of a bacterial food source and bovine serum albumin (BSA) were investigated. METHODS: Four brands of multipurpose solutions and a hydrogen peroxide disinfecting system (Oxysept) for soft CLs, and four disinfecting solutions for Rigid Gas Permeable (RGP) lenses were tested against three species of Acanthamoeba. Page's amoebic saline was included as a negative control and standard solutions of disinfecting agents, 6% hydrogen peroxide and 0.5% chlorhexidine, as positive controls. The effects of the presence of Pseudomonas aeruginosa and BSA on effectiveness were assessed. RESULTS: None of the CL solutions tested achieved a 1-log reduction in viability of all three Acanthamoeba species within the manufacturer's recommended disinfection times. The presence of P. aeruginosa did not significantly affect disinfecting capacity of multipurpose solution solutions but reduced activity of RGP solutions and the hydrogen peroxide system. BSA reduced trophozoicidal activity of all solutions. Bland and Altman analysis showed good agreement between Vi-cell and hemacytometer. CONCLUSIONS: The Vi-Cell analyzer offers a simple and effective method of determining amoebicidal activity. Our results show that the CL solutions tested could not satisfactorily kill Acanthamoeba.

Utility of real-time polymerase chain reaction in diagnosing and treating acanthamoeba keratitis.

PURPOSE: Using real-time polymerase chain reaction (PCR), we detected Acanthamoeba and monitored the changes in Acanthamoeba DNA copy number over the treatment course in patients suspected of Acanthamoeba keratitis (AK). METHODS: Subjects were 6 patients (average age, 26.2 years) suspected of AK at the Kinki University Outpatient Clinic. For detection of Acanthamoeba, patients' corneal scrapings were collected for smear analysis, culture, and real-time PCR. After the diagnosis of AK was confirmed, treatment was initiated based on the quantitative result of the real-time PCR. RESULTS: Both the smear and culture were positive for Acanthamoeba in 4 cases and negative in 2 cases (agreement in 3 cases and disagreement in 2 cases). By real-time PCR, all 6 cases were positive for Acanthamoeba with an average DNA copy number of 4.8 ± 9.1 × 10 copies per sample. We further monitored the variation in the Acanthamoeba DNA copy number over the treatment course and successfully treated all the patients. DNA copy number provided a parallel with other clinical features of AK. CONCLUSIONS: Real-time PCR can be a useful method for a rapid and precise diagnosis of AK. Moreover, utility of the Acanthamoeba DNA copy number obtained by real-time PCR can help ophthalmologists in making the best treatment decision.