RESUMO
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.
Assuntos
Acanthamoeba , Microscopia Eletrônica de Varredura , Acanthamoeba/fisiologia , Acanthamoeba/ultraestrutura , Esclera , Humanos , Lentes de Contato Hidrofílicas/parasitologia , Adesão Celular/fisiologia , Lentes de Contato/parasitologia , Trofozoítos/ultraestrutura , Trofozoítos/fisiologia , Hidrogéis , AnimaisRESUMO
Microbial keratitis is a devastating disease that can cause eye damage and blindness and can be the result of infections by several common ocular pathogens. Importantly, some of these pathogens, such as Acanthamoeba, are particularly unsusceptible to biocides in common contact lens care solutions. Therefore, the disinfection efficacy of preservative-free (PF) disinfection systems against bacteria, fungi, and Acanthamoeba trophozoites and cysts should be assessed as products with the most potential to be efficacious against resistant organisms. PF disinfection systems were analyzed for antimicrobial efficacy. These were the one-step (hydrogen peroxide-based) Clear Care and Clear Care Plus systems and the two-step (povidone-iodine-based) Cleadew system. Stand-alone challenges using bacteria, fungi, and Acanthamoeba were prepared according to the International Standards Organization method 14729. These same challenges were also conducted in the presence of the following contact lenses: Boston RGP, Acuvue Oasys, Biofinity, Ultra, and 2-week PremiO. All challenges were performed at the manufacturer's recommended disinfection time. All preservative-free disinfection systems demonstrated similarly high rates of antimicrobial efficacy when challenged with bacteria or fungi, with or without lenses. However, both Clear Care and Clear Care Plus demonstrated significantly greater disinfection efficacy against Acanthamoeba trophozoites and cysts, with and without lenses (P < 0.05). Cleadew efficacy was impacted by the addition of contact lenses, whereas Clear Care/Clear Care Plus maintained similar efficacies in the absence or presence of lenses. While both hydrogen peroxide and povidone-iodine are highly effective against bacteria and fungi, hydrogen peroxide maintains significantly greater disinfection capabilities than povidone-iodine against all forms of Acanthamoeba. IMPORTANCE Understanding the most efficacious products will allow clinicians to best communicate to patients and consumers the safest products on the market to reduce adverse events, including microbial keratitis, during contact lens use.
Assuntos
Anti-Infecciosos/farmacologia , Soluções para Lentes de Contato/farmacologia , Desinfecção/métodos , Oftalmopatias/prevenção & controle , Acanthamoeba/efeitos dos fármacos , Bactérias/efeitos dos fármacos , Lentes de Contato/microbiologia , Lentes de Contato/parasitologia , Desinfecção/instrumentação , Oftalmopatias/microbiologia , Oftalmopatias/parasitologia , Fungos/efeitos dos fármacos , Humanos , Peróxido de Hidrogênio/farmacologia , Povidona-Iodo/farmacologiaRESUMO
PURPOSE: The aim of this study was to determine the impact of Acanthamoeba keratitis (AK) caused by contact lens (CL) use on vision-related quality of life (VRQOL) and the sociodemographic factors and disease outcome associated with VRQOL. METHODS: Sixty-one CL-associated AK cases and 59 asymptomatic CL wearers (mean age ±SD 39.4 ± 16.5 vs. 45.5 ± 15.2 yrs, P = 0.04) were recruited from Moorfields Eye Hospital and Institute for Optometry, London. AK cases were surveyed during active disease and were stratified into "poor" and "good" outcomes based on clinical features. VRQOL was measured using Rasch-transformed scores from the Emotional, Mobility, and Reading domains of the 32-item Impact of Visual Impairment questionnaire. AK cases were compared with controls and "poor" outcomes compared with "good" with multivariable linear regression. Multivariable linear regression models were also used to identify the sociodemographic factors and disease outcome associated with VRQOL. RESULTS: AK was associated with significant and substantial reductions in all 3 evaluated domains of VRQOL (Reading -59.6%, Mobility -59.8%, and Emotional -66.2%) compared with controls, independent of sociodemographic factors. Patients with AK who experienced poor outcomes, those who were of British White race (compared with all other races) and female, had lower VRQOL scores across all domains. Patients with AK with lower incomes scored worse on Reading and Mobility domains, whereas those with lower education had poorer Emotional scores. CONCLUSIONS: AK has a considerable detrimental impact on VRQOL. Clinicians should consider the importance of referring patients with AK for rehabilitative support and counseling as part of active disease management.
Assuntos
Ceratite por Acanthamoeba/psicologia , Acanthamoeba/isolamento & purificação , Lentes de Contato/efeitos adversos , Infecções Oculares Parasitárias/psicologia , Qualidade de Vida , Acuidade Visual , Ceratite por Acanthamoeba/parasitologia , Ceratite por Acanthamoeba/fisiopatologia , Adulto , Estudos de Casos e Controles , Lentes de Contato/parasitologia , Córnea/parasitologia , Infecções Oculares Parasitárias/parasitologia , Infecções Oculares Parasitárias/fisiopatologia , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de RiscoRESUMO
Contact lens usage has contributed to increased incidence rates of Acanthamoeba keratitis (AK), a serious corneal infection that can lead to blindness. Since symptoms associated with AK closely resemble those incurred by bacterial or fungal keratitis, developing a diagnostic method enabling rapid detection with a high degree of Acanthamoeba-specificity would be beneficial. Here, we produced a polyclonal antibody targeting the carboxylesterase (CE) superfamily protein secreted by the pathogenic Acanthamoeba and evaluated its diagnostic potential. Western blot analysis revealed that the CE antibody specifically interacts with the cell lysates and conditioned media of pathogenic Acanthamoeba, which were not observed from the cell lysates and conditioned media of human corneal epithelial (HCE) cells, Fusarium solani, Staphylococcus aureus, and Pseudomonas aeruginosa. High titers of A. castellanii-specific antibody production were confirmed sera of immunized mice via ELISA, and these antibodies were capable of detecting A. castellanii from the cell lysates and their conditioned media. The specificity of the CE antibody was further confirmed on A. castellanii trophozoites and cysts co-cultured with HCE cells, F. solani, S. aureus, and P. aeruginosa using immunocytochemistry. Additionally, the CE antibody produced in this study successfully interacted with 7 different Acanthamoeba species. Our findings demonstrate that the polyclonal CE antibody specifically detects multiple species belong to the genus Acanthamoeba, thus highlighting its potential as AK diagnostic tool.
Assuntos
Ceratite por Acanthamoeba/diagnóstico , Acanthamoeba/imunologia , Anticorpos Antiprotozoários/análise , Carboxilesterase/imunologia , Meios de Cultivo Condicionados/metabolismo , Epitélio Corneano/citologia , Acanthamoeba/classificação , Acanthamoeba/crescimento & desenvolvimento , Acanthamoeba/isolamento & purificação , Animais , Anticorpos Antiprotozoários/sangue , Especificidade de Anticorpos , Carboxilesterase/administração & dosagem , Carboxilesterase/genética , Linhagem Celular , Células Cultivadas , Lentes de Contato/parasitologia , Diagnóstico Precoce , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Células Epiteliais/parasitologia , Epitélio Corneano/metabolismo , Epitélio Corneano/parasitologia , Humanos , Imunização , Masculino , Camundongos , Proteínas de Protozoários/administração & dosagem , Proteínas de Protozoários/genética , Proteínas de Protozoários/imunologiaRESUMO
BACKGROUND: Acanthamoeba spp. are one of the free-living amoeba that spread worldwide causing keratitis. Owing to the increase in the use of lenses, whether for medical or cosmetic purposes, the incidence of disease increases every year. Contamination of the lenses with the Acanthamoeba trophozoites or cysts may lead to eye infection and cause sight-threatening keratitis in human. We isolated Acanthamoeba spp. from new lenses, used lenses, and contact lens disinfecting solutions and identified them based on morphological characteristics and molecular test. METHODS: New and used lenses and contact lens disinfecting solutions were cultured on monogenic media. Light and scanning electron microscope was used to identify Acanthamoeba spp. morphological features. Genotype identification was also evaluated using PCR sequencing of 18S rRNA gene specific primer pair JDP1 and JDP2. RESULTS: A hundred samples were examined, 29 (29%) were infected with Acanthamoeba spp. That belonged to two strains of Acanthamoeba (Acanthamoeba 41 and Acanthamoeba 68). 18S rRNA of the Acanthamoeba 41 had 99.69% sequence identity to Acanthamoeba castellanii clone HDU-JUMS-2, whereas Acanthamoeba 68 had 99.74% similar pattern to that of Acanthamoeba sp. isolate T4 clone ac2t4 that are morphologically identified as Acanthamoeba polyphaga. The obtained data revealed that the isolated strains belong to T4 genotype that was evolutionarily similar to strains isolated in Iran. CONCLUSIONS: Cosmetic lenses and disinfectant solutions are a major transmissible mode for infection. This genotype is common as the cause of Acanthamoeba keratitis. To avoid infection, care must be taken to clean the lenses and their preservative solutions and prevent contamination with the parasite.
Assuntos
Acanthamoeba/classificação , Soluções para Lentes de Contato/análise , Lentes de Contato/parasitologia , Análise de Sequência de DNA/métodos , Acanthamoeba/genética , Acanthamoeba/isolamento & purificação , Cosméticos , DNA Ribossômico/genética , Contaminação de Medicamentos , Egito , Humanos , Irã (Geográfico) , Microscopia , Microscopia Eletrônica de Varredura , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 18S/genéticaRESUMO
SIGNIFICANCE: This study provides insight into the current recommendations, clinical behaviors, and risk assessments of eye care professionals on the topic of rigid contact lens exposure to tap water. This knowledge may motivate professional organizations to develop initiatives to educate eye care professionals on the dangers of contact lens exposure to water. PURPOSE: The purpose of this study was to investigate the practice patterns and risk perceptions of eye care professionals regarding gas-permeable contact lens exposure to tap water. METHODS: A branched-logic survey was started by 320 clinicians, vision scientists, and industry personnel and was fully completed by 272 participants. The survey queried participants about their current practices, recommendations, and perceptions of risk regarding exposure of gas-permeable contact lenses to tap water. RESULTS: Of those who prescribe gas-permeable contact lenses, 57.4% reported rinsing lenses with tap water, whereas only 32.7% reported engaging in this same behavior in front of patients. Of those who reported never rinsing lenses with tap water, 85.6% indicated that rinsing lenses with water increases infection risk, whereas 52.1% of those who rinse lenses with water reported the same perceived risk (P < .001). Of all participants, 60% indicated that wearing contact lenses while showering increases infection risk, and 50.2% of all participants advised patients to avoid this behavior. CONCLUSIONS: A discrepancy exists between perceptions of risk concerning exposure of contact lenses to water and both clinical practices and patient education provided by professionals.
Assuntos
Lentes de Contato/estatística & dados numéricos , Infecções Oculares/epidemiologia , Comportamentos Relacionados com a Saúde , Conhecimentos, Atitudes e Prática em Saúde , Prática Profissional/estatística & dados numéricos , Água/efeitos adversos , Soluções para Lentes de Contato , Lentes de Contato/microbiologia , Lentes de Contato/parasitologia , Infecções Oculares/prevenção & controle , Feminino , Inquéritos Epidemiológicos , Humanos , Masculino , Ajuste de PróteseRESUMO
Acanthamoeba keratitis is a sight-endangering eye infection, and causative organism Acanthamoeba presents a significant concern to public health, given escalation of contact lens wearers. Contemporary therapy is burdensome, necessitating prompt diagnosis and aggressive treatment. None of the contact lens disinfectants (local and international) can eradicate Acanthamoeba effectively. Using a range of compounds targeting cellulose, ion channels, and biochemical pathways, we employed bioassay-guided testing to determine their anti-amoebic effects. The results indicated that acarbose, indaziflam, terbuthylazine, glimepiride, inositol, vildagliptin and repaglinide showed anti-amoebic effects. Compounds showed minimal toxicity on human cells. Therefore, effects of the evaluated compounds after conjugation with nanoparticles should certainly be the subject of future studies and will likely lead to promising leads for potential applications.
Assuntos
Ceratite por Acanthamoeba/tratamento farmacológico , Ceratite por Acanthamoeba/parasitologia , Acanthamoeba castellanii/efeitos dos fármacos , Antiprotozoários/farmacologia , Lentes de Contato/parasitologia , Acarbose/farmacologia , Carbamatos/farmacologia , Linhagem Celular , Soluções para Lentes de Contato/farmacologia , Lentes de Contato/efeitos adversos , Células HaCaT , Humanos , Indenos/farmacologia , Inositol/farmacologia , Nanopartículas , Piperidinas/farmacologia , Compostos de Sulfonilureia/farmacologia , Triazinas/farmacologia , Vildagliptina/farmacologiaRESUMO
Acanthamoeba keratitis (AK) is a rare protozoal infection of the cornea. At least eight species of Acanthamoeba are known to cause this sight-threatening disease of the ocular surface. Acanthamoeba spp. exist in a wide array of niches ranging from thermal springs to under ice and every conceivable habitat in between. Contact lens wear is the leading risk factor for AK and is practiced by over 30 million individuals in the United States, yet the incidence of AK is less than 33 cases per one million contact lens wearers. Serological studies have reported that 90%-100% of individuals with no history of AK possess antibodies specific for Acanthamoeba antigens indicating that exposure to this organism is commonplace, yet disease is remarkably rare. Animal studies have shed light on the pathobiology and immunobiology of AK and indicate that a constellation of factors including the ocular surface microbiome and the microbiome of Acanthamoeba itself contribute to the pathogenesis of AK. Interesting, secretory antibodies produced by the adaptive immune response can prevent the initiation of corneal infection, but once Acanthamoeba trophozoites breach the corneal epithelium the adaptive immune system is helpless in altering the course of AK. It has been almost 50 years since AK was first described, yet many questions remain unanswered about this curious and enigmatic disease of the ocular surface.
Assuntos
Ceratite por Acanthamoeba/diagnóstico , Acanthamoeba/imunologia , Lentes de Contato/parasitologia , Córnea/patologia , Infecções Oculares Parasitárias/diagnóstico , Ceratite por Acanthamoeba/parasitologia , Animais , Anticorpos Antiprotozoários/imunologia , Lentes de Contato/efeitos adversos , Córnea/parasitologia , Infecções Oculares Parasitárias/parasitologia , Humanos , Fatores de RiscoRESUMO
Curcuma longa and Curcumin have been documented to have a wide spectrum of pharmacological effects, including anti-Acanthamoeba activity. Hence, this study sought to explore the anti-adhesion activity of C. longa extract and Curcumin against Acanthamoeba triangularis trophozoites and cysts in plastic and contact lenses. Our results showed that C. longa extract and Curcumin significantly inhibited the adhesion of A. triangularis trophozoites and cysts to the plastic surface, as investigated by the crystal violet assay (P < 0.05). Also, an 80-90% decrease in adhesion of trophozoites and cysts to the plastic surface was detected following the treatment with C. longa extract and Curcumin at 1/2 × MIC, compared to the control. In the contact lens model, approximately 1 log cells/mL of the trophozoites and cysts was reduced when the cells were treated with Curcumin, when compared to the control. Pre-treatment of the plastic surface with Curcumin at 1/2-MIC reduced 60% and 90% of the adhesion of trophozoites and cysts, respectively. The reduction in 1 Log cells/mL of the adhesion of A. triangularis trophozoites was observed when lenses were pre-treated with both the extract and Curcumin. Base on the results obtained from this study, A. triangularis trophozoites treated with C. longa extract and Curcumin have lost strong acanthopodia, thorn-like projection pseudopodia observed by scanning electron microscope. This study also revealed the therapeutic potentials of C. longa extract and Curcumin, as such, have promising anti-adhesive potential that can be used in the management/prevention of A. triangularis adhesion to contact lenses.
Assuntos
Acanthamoeba , Lentes de Contato , Curcumina , Extratos Vegetais , Acanthamoeba/efeitos dos fármacos , Animais , Lentes de Contato/parasitologia , Curcuma/química , Curcumina/farmacologia , Extratos Vegetais/farmacologia , Plásticos , Poliestirenos , Rizoma/química , TrofozoítosRESUMO
PURPOSE: To determine whether Acanthamoeba keratitis (AK) patients have higher rates of Acanthamoeba and free-living amoeba (FLA) colonising domestic sinks than control contact lens (CL) wearers, and whether these isolates are genetically similar to the corneal isolates from their CL associated AK. METHODS: 129 AK patients from Moorefield Eye Hospital, London and 64 control CL wearers from the Institute of Optometry were included in this study. The participants self-collected home kitchen and bathroom samples from tap-spouts, overflows and drains using an instructional kit. The samples were cultured by inoculating onto a non-nutrient agar plate seeded with Escherichia coli, incubated at 32°C and examined for amoebae by microscopy for up to 2 weeks. Partial sequences of mitochondrial cytochrome oxidase genes (coxA) of Acanthamoeba isolates from four AK patients were compared to Acanthamoeba isolated from the patient's home. The association between sampling sites was analysed with the chi-square test. RESULTS: A total of 513 samples from AK patients and 189 from CL controls were collected. The yield of FLA was significantly greater in patients' bathrooms (72.1%) than CL controls' bathrooms (53.4%) (p<0.05). Spouts (kitchen 6.7%, bathroom 11%) had the lowest rate of Acanthamoeba isolation compared to drains (kitchen 18.2%, bathroom 27.9%) and overflow (kitchen 39.1%, bathroom 25.9%) either in kitchens or bathrooms (p<0.05). There was no statistically significant difference between the average prevalence of Acanthamoeba in all three sample sites in kitchens (16.9%) compared to all three sample sites in bathrooms (21.5%) and no association for Acanthamoeba prevalence between AK patients and CL controls. All four corneal isolates had the same coxA sequence as at least one domestic water isolate from the patients' sink of the kitchen and the bathroom. CONCLUSION: The prevalence of Acanthamoeba and FLA was high in UK homes. FLA colonisation was higher in AK patients compared to controls but the prevalence of Acanthamoeba between AK patients and CL controls domestic sinks was similar. This study confirms that domestic water isolates are probably the source of AK infection. Advice about avoiding water contact when using CL's should be mandatory.
Assuntos
Ceratite por Acanthamoeba/epidemiologia , Ceratite por Acanthamoeba/parasitologia , Acanthamoeba/isolamento & purificação , Acanthamoeba/genética , Acanthamoeba/patogenicidade , Amebozoários/isolamento & purificação , Estudos de Casos e Controles , Lentes de Contato/efeitos adversos , Lentes de Contato/parasitologia , Suscetibilidade a Doenças , Meio Ambiente , Habitação , Humanos , Londres/epidemiologia , Fatores de Risco , Engenharia Sanitária , Água/parasitologiaRESUMO
BACKGROUND: To report the clinical and microbiological features of Acanthamoeba keratitis (AK) related to contact lens use in a tertiary hospital in China. METHODS: In this retrospective study, the medical results of 61 cases of AK related to contact lens use from January 2000 to December 2017 were reviewed. The data included patients' demographics, lens type, history, risk factors, disease stages, corneal scraping and culture reports, and treatments. Moreover, genotypic identification of some of the isolates was carried out with a PCR assay and sequence analysis of the 18S ribosomal DNA gene. RESULTS: There were 64 eyes included in the study. A total of 32.8% of the patients wore soft contact lenses, and 67.2% of patients used overnight orthokeratology. In the cases (20 eyes) in the early stage, 65% (13 eyes) had positive results according to Giemsa-stained smears, and 0.9% sodium chloride (NaCl) wet mounts revealed trophozoites in 7 eyes (35%). Six eyes (30%) were diagnosed by confocal microscopy combined with clinical signs. In the orthokeratology patients, 87.8% (36/41) rinsed their lenses and/or cases with tap water; 55% of soft-lens wearers wore their lenses while showering. The genotype of 9 isolates was determined, and all the strains belonged to genotype T4. In the orthokeratology group, the number of patients who required therapeutic penetrating keratoplasty after 2005 was less than that before 2005 (chi-square test, χ2 = 4.209, P = 0.04). CONCLUSIONS: More than two-thirds of the cases were associated with orthokeratology. Examinations with Giemsa-stained smears, 0.9% NaCl wet mounts and confocal microscopy should be performed for patients who are highly suspected of having early-stage AK to help with early diagnosis. In the orthokeratology group, the rate of therapeutic keratoplasty after 2005 was less than that before 2005.
Assuntos
Ceratite por Acanthamoeba/etiologia , Lentes de Contato/parasitologia , Úlcera da Córnea/etiologia , Infecções Oculares Parasitárias/etiologia , Acanthamoeba/genética , Acanthamoeba/isolamento & purificação , Ceratite por Acanthamoeba/diagnóstico , Ceratite por Acanthamoeba/terapia , Adolescente , Adulto , Anti-Infecciosos Locais/uso terapêutico , Biguanidas/uso terapêutico , Criança , China , Clorexidina/uso terapêutico , Úlcera da Córnea/diagnóstico , Úlcera da Córnea/terapia , DNA de Protozoário/genética , Desinfetantes/uso terapêutico , Combinação de Medicamentos , Infecções Oculares Parasitárias/diagnóstico , Infecções Oculares Parasitárias/terapia , Feminino , Humanos , Ceratoplastia Penetrante , Masculino , Microscopia Confocal , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , RNA Ribossômico 18S/genética , Estudos Retrospectivos , Centros de Atenção Terciária , Adulto JovemRESUMO
PURPOSE: To compare the infectious contact lens-related corneal ulcer (CLRU) and non-CLRU cases at Saint Louis University. METHODS: Retrospective review of corneal ulcer cases identified by search of the ophthalmology and microbiology department databases between 1999 and 2016. RESULTS: Six hundred seventy-seven cases of corneal ulcers were identified, of which 46% were CLRU. CLRU cases were seen more commonly in younger patients (P<0.001) and women (P=0.03) than non-CLRU cases. Many of the infections were vision-threatening as defined by central/paracentral location (73% CLRU and 71% non-CLRU [P=0.60]) and large size of ulcer >2 mm in 36% CLRU and 51% non-CLRU (P=0.002). Causative pathogen in cultured CLRU was predominately Pseudomonas species (44%, P<0.001 vs. the non-CLRU group), other gram-negative (6%), gram-positive (33%), fungi (13%), and Acanthamoeba (5%). Comparatively, cultured non-CLRU was predominately gram-positive (64%, P<0.001 vs. the CLRU group), gram-negative (26%), and fungi (11%). The combined oxacillin-resistant Staphylococcus aureus and coagulase-negative Staphylococcus isolates were 35% and 34%, respectively. Despite the progressive increase in the number of corneal ulcers seen, the annual trend for any one particular organism for either CLRU cases or non-CLRU cases did not change significantly. CONCLUSIONS: Most of the cases were non-CLRU. CLRU was disproportionately associated with Pseudomonas species and non-CLRU with Staphylococcal species. Fungal infections were predominately caused by filamentous organisms in both groups. Acanthamoeba keratitis was exclusively associated with CL use.
Assuntos
Ceratite por Acanthamoeba/parasitologia , Lentes de Contato/microbiologia , Lentes de Contato/parasitologia , Úlcera da Córnea/microbiologia , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Fúngicas/microbiologia , Infecções Oculares Parasitárias/parasitologia , Centros Médicos Acadêmicos , Ceratite por Acanthamoeba/diagnóstico , Ceratite por Acanthamoeba/tratamento farmacológico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/uso terapêutico , Antiprotozoários/uso terapêutico , Criança , Pré-Escolar , Úlcera da Córnea/diagnóstico , Úlcera da Córnea/tratamento farmacológico , Infecções Oculares Bacterianas/diagnóstico , Infecções Oculares Bacterianas/tratamento farmacológico , Infecções Oculares Fúngicas/diagnóstico , Infecções Oculares Fúngicas/tratamento farmacológico , Infecções Oculares Parasitárias/diagnóstico , Infecções Oculares Parasitárias/tratamento farmacológico , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
RESUMEN El parasitismo es uno de los fenómenos más sorprendentes de los observados en los animales. El número de organismos conocidos de vida parasitaria es muy elevado. Existe una gran cantidad de especies cuya supervivencia está relacionada con la de otras y dependen, en distinta medida, de ellas. Esta dependencia no implica que los animales parásitos sean organismos degenerados o deficientes; al contrario, a la vida parasitaria se ha llegado tras largo tiempo de evolución, en el que los parásitos han ido superando barreras y adaptándose a vivir en, o sobre sus hospedadores. Las queratitis por Acanthamoeba son infrecuentes; pero se describe un aumento a nivel mundial relacionado con el uso creciente de lentes de contacto. El diagnóstico precoz y el tratamiento adecuado deben realizarse para evitar la pérdida de la visión. Se realizó una búsqueda de artículos publicados, con el objetivo de conocer sobre la Acanthamoeba como parásito y su afectación ocular. Se utilizó la plataforma Infomed, específicamente la Biblioteca Virtual de Salud(AU)
ABSTRACT Parasitism is one of the most surprising phenomena among those occurring in animals. The number of known parasitic organisms is very high. There is a large number of species whose survival is related to that of others and depend on them to a greater or lesser degree. Such dependence does not imply that parasitic animals are either degenerate or deficient. On the contrary, parasitic life has been the result of a long process of evolution along which parasites have gradually overcome hurdles and have adapted to live in or on their hosts. Acanthamoeba keratitis is a rare disease, but a worldwide increase has been reported due to the growing use of contact lenses. Early diagnosis and appropriate treatment are required to prevent sight loss. A search was conducted for published papers with the purpose of learning about Acanthamoeba as a parasite and the related eye conditions. Use was made of the platform Infomed, specifically the Virtual Health Library(AU)
Assuntos
Humanos , Doenças Parasitárias/tratamento farmacológico , Ceratite por Acanthamoeba/epidemiologia , Transplante de Córnea/métodos , Lentes de Contato/parasitologia , Literatura de Revisão como AssuntoRESUMO
Free-living amoebae belong to the genus Acanthamoeba; can feed on microbial population by phagocytosis, and with the capability to act as a reservoir and a vehicle of microorganisms to susceptible host. Therefore, the role of endosymbiosis in the pathogenesis of Acanthamoeba is complex and not fully understood. The aim of the present study was to identify bacterial, fungal, and human adenovirus (HADV) endosymbionts as well as evaluating the endosymbionts role of such organisms in the pathogenesis of Acanthamoeba in keratitis patients living in Iran. Fifteen Acanthamoeba (T4 genotype) isolates were recovered from corneal scrapes and contact lenses of patients with keratitis. Cloning and purification was performed for all isolate. Gram staining was performed to identify bacterial endosymbionts. DNA extraction, PCR, and nested PCR was set up to identify endosymbiont of amoeba. Evaluation of pathogenicity was conducted by osmo-tolerance and thermo-tolerance assays and cell culture, and then CPE (cytopathic effect) was survey. Statistical analysis was used between Acanthamoeba associated endosymbionts and Acanthamoeba without endosymbiont at 24, 48, 72, and 96â¯h. A p valueâ¯<â¯0.05 was considered as significant, statistically. A total of 9 (60%) Acanthamoeba (T4 genotypes) isolates were successfully cloned for detecting microorganism endosymbionts. The only isolate negative for the presence of endosymbiont was ICS9. ICS7 (Pseudomonas aeruginosa, Aspergillus sp., and human adenovirus endosymbionts) and ICS2 (Escherichia coli endosymbiont) isolates were considered as Acanthamoeba associated endosymbionts. ICS7 and ICS2 isolates were highly pathogen whereas ICS9 isolate showed low pathogenicity in pathogenicity evaluated. Positive CPE for ICS7 and ICS2 isolates and negative CPE for ICS9 isolate were observed in cell culture. The average number of cells, trophozoites, and cysts among ICS7, ICS2, and ICS9 isolates at 24, 48, 72, and 96â¯h was significant. This is the first survey on microbial endosymbionts of Acanthamoeba in keratitis patients of Iran, and also the first report of Aspergillus sp, Achromobacter sp., Microbacterium sp., Brevibacillus sp, Brevundimonas sp and Mastadenovirus sp in Acanthamoeba as endosymbionts. Our study demonstrated that microbial endosymbionts can affect the pathogenicity of Acanthamoeba; however, further research is required to clarify the exact pattern of symbiosis, in order to modify treatment protocol.
Assuntos
Ceratite por Acanthamoeba/complicações , Acanthamoeba/fisiologia , Adenovírus Humanos/isolamento & purificação , Bactérias/isolamento & purificação , Fungos/isolamento & purificação , Simbiose , Acanthamoeba/isolamento & purificação , Acanthamoeba/microbiologia , Acanthamoeba/patogenicidade , Adenovírus Humanos/genética , Adenovírus Humanos/fisiologia , Animais , Bactérias/genética , Chlorocebus aethiops , Clonagem Molecular , Doenças Transmissíveis/microbiologia , Doenças Transmissíveis/transmissão , Lentes de Contato/parasitologia , Córnea/parasitologia , Reservatórios de Doenças , Fungos/genética , Humanos , Irã (Geográfico) , Reação em Cadeia da Polimerase , Células Vero , VirulênciaRESUMO
PURPOSE: To ascertain the incidence of Acanthamoeba keratitis and the coexistence of Acanthamoeba and fungi in microbial keratitis. DESIGN: Prospective cross-sectional study. METHODS: Patients presenting with stromal keratitis were additionally tested for Acanthamoeba irrespective of the clinical diagnosis. Culture positivity was the gold standard. RESULTS: Of the 401 cases included in the study, 40 were positive for Acanthamoeba (10%); of these 40, 16 were positive for both Acanthamoeba and fungi (4.5% of the study group was Acanthamoeba and fungal keratitis positive); 5 were positive for Acanthamoeba and bacteria; and 2 had triple infection with Acanthamoeba, fungi, and bacteria. Ring infiltrates and stromal edema are frequently associated with Acanthamoeba keratitis, as well as in Acanthamoeba coinfections. Ring infiltrates in particular were more frequently seen in the Acanthamoeba and fungal keratitis group (8/16) and they were often yellowish with hyphate edges (vs ring infiltrates only, which are seen in the patients with Acanthamoeba alone). Only 2 patients were contact lens wearers: however, they presented with history of trauma. CONCLUSIONS: Acanthamoeba coinfections are much more frequent and are not restricted to contact lens users. Anticipating coinfections is necessary for establishing a diagnosis as well as for appropriate and timely therapeutic interventions.
Assuntos
Ceratite por Acanthamoeba/epidemiologia , Coinfecção/epidemiologia , Úlcera da Córnea/epidemiologia , Úlcera da Córnea/parasitologia , Infecções Oculares Fúngicas/epidemiologia , Infecções Oculares Parasitárias/epidemiologia , Acanthamoeba/isolamento & purificação , Ceratite por Acanthamoeba/diagnóstico , Ceratite por Acanthamoeba/tratamento farmacológico , Ceratite por Acanthamoeba/microbiologia , Adolescente , Adulto , Antibacterianos/uso terapêutico , Antiprotozoários/uso terapêutico , Bactérias/isolamento & purificação , Coinfecção/diagnóstico , Coinfecção/tratamento farmacológico , Coinfecção/microbiologia , Lentes de Contato/microbiologia , Lentes de Contato/parasitologia , Substância Própria/microbiologia , Substância Própria/parasitologia , Úlcera da Córnea/diagnóstico , Úlcera da Córnea/microbiologia , Estudos Transversais , Infecções Oculares Bacterianas/diagnóstico , Infecções Oculares Bacterianas/tratamento farmacológico , Infecções Oculares Bacterianas/epidemiologia , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Fúngicas/diagnóstico , Infecções Oculares Fúngicas/tratamento farmacológico , Infecções Oculares Fúngicas/microbiologia , Infecções Oculares Parasitárias/diagnóstico , Infecções Oculares Parasitárias/tratamento farmacológico , Infecções Oculares Parasitárias/microbiologia , Feminino , Fungos/isolamento & purificação , Humanos , Índia/epidemiologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Adulto JovemRESUMO
Free-living amoebae of the genus Acanthamoeba are the etiological agents of cutaneous lesions, granulomatous amoebic encephalitis (GAE) and amoebic keratitis (AK), which are chronic infections with poor prognosis if not diagnosed promptly. Currently, there is no optimal therapeutic scheme to eradicate the pathologies these protozoa cause. In this study we report the morphological and molecular identification of three species of the genus Acanthamoeba, belonging to T4 group; A. polyphaga isolated from the corneal ulcer of a patient sample of AK case; A. castellanii isolated from the contact lens of an AK patient and A. palestinensis obtained from a soil sample. The in vitro activity of chlorhexidine, itraconazole and voriconazole drugs against trophic stage was also evaluated through a colorimetric assay based on the oxidation-reduction of alamar blue. The strains in the study were sensitive to the evaluated drugs; although when determining the 50% inhibitory concentration (IC50) statistically significant differences were observed. A. castellanii showed to be highly sensitive to voriconazole (0.66⯱â¯0.13⯵M) but the least sensitive to chlorhexidine and itraconazole (8.61⯱â¯1.63 and 20.14⯱â¯4.93⯵M, respectively), A. palestinensis showed the highest sensitivity to itraconazole (0.502⯱â¯0.11⯵M) and A. polyphaga expressed moderate sensitivity to chlorhexidine and itraconazole and lower sensitivity to voriconazole (10.10⯱â¯2.21⯵M). These results showed that species of the genus Acanthamoeba express different sensitivity to the tested drugs, which could explain the problems surrounding the establishment of a treatment of choice in the infections caused by these amoebae. We consider that although chlorhexidine and itraconazole show good activity on these amoebae and have been used in cases of AK in Mexico with acceptable results, voriconazole should be considered as the first therapeutic option of future Acanthamoeba infections that will be diagnosed in our country.
Assuntos
Acanthamoeba/efeitos dos fármacos , Amebíase/parasitologia , Anti-Infecciosos/farmacologia , Clorexidina/farmacologia , Itraconazol/farmacologia , Voriconazol/farmacologia , Acanthamoeba/classificação , Acanthamoeba/genética , Ceratite por Acanthamoeba/parasitologia , Amebíase/tratamento farmacológico , Lentes de Contato/parasitologia , Úlcera da Córnea/parasitologia , DNA de Protozoário/isolamento & purificação , Genótipo , Humanos , Concentração Inibidora 50 , México , Solo/parasitologiaRESUMO
Acanthamoeba keratitis (AK) is a sight-threatening corneal infection. The early symptoms include redness, pain, photophobia and intense tearing. Chronic infection usually progresses to stromal inflammation, ring ulcers, corneal opacification and hypopyon. Here we document an AK case in a high myopic 38-year-old woman from Mexico City, with a history of wearing contact lenses while swimming. Corneal scrapes cultures were positive only for amoebae, consequently a treatment including netilmicin 0.3% and oral itraconazole 100 mg/12â¯h was prescribed. The infection was resolved after 8 months, leaving a slight leucoma outside the visual axis, with a visual acuity of 20/150. In the laboratory, the amoebic isolate was axenized in PYG medium, with an optimal growth at 30⯰C, and was identified morphologically as Acanthamoeba polyphaga according to the taxonomic criteria of Page (1988) and placed in the T4 group by genotyping. The virulence of this strain (40%) was determined by intranasal inoculation of 1â¯×â¯106/20⯵l trophozoites in BALB/c mice recovering from brain, proving their invasion ability and by the interaction with monolayers of epithelial cells of the established MDCK line of canine kidney origin (1:2 ratio of interaction), at 1, 3, 6, 8 and 24â¯h; trophozoites migrated to cell junctions inducing few lytic zones. In addition to the biological characterization, in vitro drug sensitivity tests were performed using chlorhexidine, itraconazole, netilmicin and voriconazole. Results revealed that voriconazole was the most effective compound. A. polyphaga remains as one of the most frequently isolated species producing AK. The treatment of AK case using netilmicin and oral itraconazole solved the disease, but the healing process was wide-ranging (8 months). The use of voriconazole and chlorhexidine may be an alternative treatment of future AK cases in Mexico.
Assuntos
Ceratite por Acanthamoeba/parasitologia , Acanthamoeba/efeitos dos fármacos , Anti-Infecciosos/administração & dosagem , Acanthamoeba/isolamento & purificação , Ceratite por Acanthamoeba/tratamento farmacológico , Adulto , Animais , Antibacterianos/administração & dosagem , Antibacterianos/farmacologia , Anti-Infecciosos/farmacologia , Antifúngicos/administração & dosagem , Antifúngicos/farmacologia , Clorexidina/farmacologia , Lentes de Contato/efeitos adversos , Lentes de Contato/parasitologia , Cães , Feminino , Humanos , Concentração Inibidora 50 , Itraconazol/administração & dosagem , Itraconazol/farmacologia , Células Madin Darby de Rim Canino , México , Camundongos , Camundongos Endogâmicos BALB C , Midriáticos/administração & dosagem , Netilmicina/administração & dosagem , Netilmicina/farmacologia , Testes de Sensibilidade Parasitária , Fenilefrina/administração & dosagem , Tropicamida/administração & dosagem , Voriconazol/farmacologiaAssuntos
Ceratite por Acanthamoeba/prevenção & controle , Pesquisa Biomédica/tendências , Infecções Oculares Parasitárias/prevenção & controle , Conhecimentos, Atitudes e Prática em Saúde , Testes Imediatos/tendências , Ceratite por Acanthamoeba/tratamento farmacológico , Ceratite por Acanthamoeba/epidemiologia , Ceratite por Acanthamoeba/parasitologia , Animais , Antiparasitários/uso terapêutico , Conscientização , Lentes de Contato/parasitologia , Infecções Oculares Parasitárias/tratamento farmacológico , Infecções Oculares Parasitárias/epidemiologia , Infecções Oculares Parasitárias/parasitologia , Humanos , Reino Unido/epidemiologiaRESUMO
Multipurpose contact lens disinfecting solutions (MPDS) are widely used to cleanse and disinfect microorganisms. However, disinfection efficacy of these MPDS against Acanthamoeba cyst remain insufficient. 2, 6-dichlorobenzonitrile (DCB), a cellulose synthesis inhibitor, is capable of increasing the amoebical effect against Acanthamoeba by inhibiting its encystation. In this study, we investigated the possibility of DCB as a disinfecting agent to improve the amoebicidal activity of MPDS against Acanthamoeba cyst. Eight commercial MPDS (from a to h) were assessed, all of which displayed insufficient amoebicidal activity against the mature cysts. Solution e, f, and h showed strong amoebicidal effect on the immature cysts. Amoebicidal efficacy against mature cysts remained inadequate even when the 8 MPDS were combined with 100 µM DCB. However, 4 kinds of MPDS (solution d, e, f, and h) including 100 µM DCB demonstrated strong amoebicidal activity against the immature cysts. The amoebicidal activity of solution d was increased by addition of DCB. Cytotoxicity was absent in human corneal epithelial cells treated with either DCB or mixture of DCB with MPDS. These results suggested that DCB can enhance the amoebicical activity of MPDS against Acanthamoeba immature cyst in vitro.
Assuntos
Acanthamoeba/efeitos dos fármacos , Amebicidas/farmacologia , Soluções para Lentes de Contato/farmacologia , Nitrilas/farmacologia , Acanthamoeba/metabolismo , Células Cultivadas , Celulose/metabolismo , Soluções para Lentes de Contato/efeitos adversos , Lentes de Contato/parasitologia , Células Epiteliais/efeitos dos fármacos , Epitélio Corneano/efeitos dos fármacos , Humanos , Nitrilas/efeitos adversos , Encistamento de Parasitas/efeitos dos fármacosRESUMO
We report an Acanthamoeba keratitis case associated with the use of contact lens in a 28-year-old female from Brasília, Brazil. Samples from corneal scraping and contact lens case were used for culture establishment, PCR amplification, and partial sequencing (fragments of ~400kb) of small subunit rDNA; both culture and PCR were positive. The sequence analyses of the cornea and of isolates from the contact lens case showed similarity with the T4 genotype. To the best of our knowledge, this is the first report of T4 Acanthamoeba keratitis case from the Midwest region of Brazil.