RARE DISEASE
Amoebic keratitis
Amoebic keratitis
Amoebic keratitis
Drug discovery
6
drugs
With orphan designations
Overview
Acanthamoeba keratitis (AK) is a rare, sight-threatening corneal infection caused by Acanthamoeba protozoa. Predominantly affecting contact lens wearers, it arises from exposure to contaminated water or poor lens hygiene. Early symptoms mimic bacterial/viral keratitis, delaying diagnosis. Treatment involves dual topical therapy (chlorhexidine/polyhexamethylene biguanide + propamidine) targeting trophozoites and cysts, often requiring months of therapy. Severe cases may necessitate corneal transplant. Prognosis depends on early intervention but carries risks of permanent vision loss [1][3][12].
Burden
Global incidence: ~2.34 cases/million population, rising steadily due to contact lens misuse [2][7][12].
High morbidity: 41% risk of legal blindness in advanced cases; prolonged treatment (6–12 months) common [3][16][17].
Economic cost: $175M/year in U.S. healthcare expenditures, including antimicrobial prescriptions and surgical interventions [4][9][17].
Therapies
First-line: Dual topical therapy with biguanides (chlorhexidine 0.02%, PHMB 0.02–0.06%) and diamidines (propamidine 0.1%) hourly initially, tapered over months [1][3][8].
Adjunctive: Epithelial debridement, oral miltefosine for refractory cases, and cautious steroid use for inflammation [3][8][12].
Surgical: Penetrating keratoplasty reserved for perforation or scarring post-infection control [1][8].
Categories: rare infectious diseases, rare ophthalmic disorders
Research Papers
1,056 drug discovery papers about Amoebic keratitis, with 4 first-in-class and 9 next-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
1,056 drug discovery papers about Amoebic keratitis, with 4 first-in-class and 9 next-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
categories:
Small molecules
small molecules
2026-09-03 | Clinical and Microbiological Comparison of Acanthamoeba Keratitis According to the Existence of Hypopyon.
To compare the clinical outcomes of patients with Acanthamoeba keratitis (AK) according to the existence of hypopyon. A retrospective analysis of microbiologically confirmed AK cases over the past 15 years was conducted. Nine eyes with hypopyon (group 1) and 29 eyes without hypopyon (group 2) were analyzed to study the differences in these 2 groups of AK eyes. The cases were diagnosed using conventional microscopic analysis with staining techniques with smear and cultural methods and were treated according to diagnosis. There was no difference in the demographic profiles and duration of symptoms before first visit in the 2 groups. The presenting corneal features, presenting best-corrected visual acuity, and final best-corrected visual acuity were significantly different between groups (P = 0.042, P = 0.002, P = 0.001, respectively). There were significant differences between groups in microbiologic diagnosis (P < 0.001): bacteria with Acanthamoeba (0/9 in group 1 vs. 4/29 in group 2), fungus with Acanthamoeba (7/9 in group 1 vs. 0/29 in group 2), and Acanthamoeba only (2/9 in group 1 vs. 25/29 in group 2). The eyes with fungus and AK coinfection showed worse visual prognosis. AK with hypopyon can be a representative sign of fungal coinfection, which has a trend of a worse visual prognosis. It should immediately alert clinicians to initiate early antifungals and antiamoebics.
2026-09-02 | Anti-Acanthamoeba Activities of Silica Nanoformulated Quercetin Compound Derived From Annona muricata L. Leaves.
Acanthamoeba spp., free-living amoebae (FLA), are the causative agents of Acanthamoeba keratitis (AK) and granulomatous amoebic encephalitis (GAE). These diseases have limited treatment options; therefore, drug discovery remains a critical priority. This study evaluated the leaf extract of Annona muricata L., which contains quercetin, formulated in silica nanoparticles against Acanthamoeba species. The anti-Acanthamoeba effect of the nanoformulation was determined by its MIC and percentage viability. Scanning electron microscopy confirmed the nanocompound-directed effect on trophozoites and cysts. The nanoquercetin exhibited anti-Acanthamoeba activity, showing greater effectiveness against the trophozoites of Acanthamoeba polyphaga ATCC 30461 (AP) (MIC > 2 mg/mL) compared to cysts (MIC > 8 mg/mL) and against A. triangularis WU19001 (AT) trophozoites (MIC > 4 mg/mL) compared to cysts (MIC > 8 mg/mL). It also showed reduced activity against Acanthamoeba castellanii ATCC 30010 (AC3) trophozoites (MIC > 4 mg/mL) compared to cysts (MIC > 8 mg/mL) and A. castellanii ATCC50739 (AC5) trophozoites (MIC > 4 mg/mL) compared to cysts (MIC > 8 mg/mL). The anti-Acanthamoeba activity of A. muricata on all four species was compared to chlorhexidine, which had MIC values of 0.008 mg/mL for AC3 and AT trophozoites and 0.016 mg/mL for AC5 and AP trophozoites. For cysts, chlorhexidine exhibited MIC values of > 0.0128 mg/mL for AC3 and AP, 0.032 mg/mL for AC5, and 0.064 mg/mL for AT. All MIC values demonstrated inhibition of Acanthamoeba species with reduced viability exceeding 90%. SEM findings revealed morphological changes in Acanthamoeba, indicating parasitic inhibition. Quercetin-loaded silica nanoparticles demonstrated anti-Acanthamoeba activity with low cytotoxicity. This indicates the potential of quercetin as a nano-enhanced natural therapeutic or contact lens-care agent. This study is limited by its in vitro design, small parasite strain panel, and potential nanoparticle variability. This highlights the need for in vivo validation, ocular/contact lens model testing, and formulation optimization for improved cysticidal efficacy.
2026-08-28 | An expert's guide to Acanthamoeba keratitis pharmacology.
Acanthamoeba keratitis (AK) is a rare but sight-threatening corneal infection, increasingly associated with contact lens wear, whose treatment remains constrained by the lack of agents that are both cysticidal and well tolerated by the cornea. This review appraises the current pharmacological landscape and the obstacles impeding therapeutic progress. PubMed, Scopus, Google Scholar, and Web of Science were searched for articles published between January 2000 and June 2026 using combinations of the terms 'Acanthamoeba keratitis,' 'treatment,' 'pharmacology,' 'amoebicidal' and individual drug names; reference lists were hand-searched. We provide an overview of the epidemiology, biology, life cycle and pathogenic mechanisms of AK. We then critically evaluate current clinical management strategies and their inherent limitations before providing a comprehensive synthesis of recent in vitro and in vivo investigations to identify effective drugs that target both the cyst and trophozoite stages with low cytotoxicity. Prolonged, intensive topical regimens impose a substantial burden that drives poor adherence and relapse, and standardized protocol-based dosing offers the most immediate practical gain. The field now requires agents combining potent cysticidal activity with corneal biocompatibility, alongside standardized assays and validated ex vivo models, so that promising laboratory activity translates into genuine clinical benefit.
2026-08-27 | Oxidative Stress and Redox Imbalance in Acanthamoeba Keratitis: Host-Parasite Crosstalk and Therapeutic Perspectives.
Acanthamoeba keratitis is a severe, sight-threatening corneal infection caused by free-living amoebae widely distributed in soil, natural waters, and domestic or recreational water systems. Following entry into the corneal microenvironment, disease develops within a redox conflict in which host-derived reactive oxygen species (ROS) and reactive nitrogen species (RNS) may contribute to parasite control while simultaneously promoting inflammation and tissue injury. Acanthamoeba responds through an adaptable defense network involving superoxide dismutases, catalase, thioredoxin- and glutathione-dependent systems, peroxiredoxins, and mitochondrial mechanisms. However, most mechanistic evidence derives from in vitro trophozoite studies, whereas the contribution of individual redox pathways to mature cyst survival and persistence in the infected cornea remains poorly defined. This review critically integrates evidence on host oxidative and nitrosative responses, parasite antioxidant adaptation, bidirectional redox crosstalk, and redox-modulating therapeutic strategies. Current data support neither indiscriminate suppression nor uncontrolled amplification of reactive species. Instead, future interventions should achieve spatially, temporally, and stage-specifically controlled redox modulation that weakens parasite defenses while preserving corneal integrity and essential innate immune functions. Progress will require causal validation of redox targets, separate assessment of trophozoites and mature cysts, long-term excystation and regrowth assays, evaluation across clinical isolates and genotypes, and parallel assessment of host-cell toxicity in clinically relevant corneal models.
2026-08-20 | Benzophenone Derivatives as a New Class of Anti-Infectives Against Acanthamoeba spp.
The rare but sight-threatening ocular disease Acanthamoeba keratitis represents a therapeutic challenge due to its causative agents, Acanthamoeba spp., being able to penetrate the corneal epithelium and persist in the corneal stroma as dormant and highly resistant cysts. To combat the scarcity and limitations of current treatment options, we developed a new class of anti-infective agents against Acanthamoeba spp. based on Ro 48-8071 (3), a known inhibitor of the human oxidosqualene cyclase (hOSC). The compound design was guided by docking studies using a homology model of the cycloartenol synthase (CAS), which is the homolog of hOSC found in Acanthamoeba spp. and essential for the parasite's steroid biosynthesis. Among the synthesized Ro 48-8071 analogues, MSHG19 (4f) evoked the most potent activity against Acanthamoeba hatchetti leading to total eradication of trophozoites and cysts at single-digit micromolar concentrations, whereas having no effects within this concentration range on the viability of human corneal epithelial cells. With our initial set of synthesized compounds, we established a first structure-activity relationship model, which nicely aligns with the predicted binding mode of 4f to CAS. Overall, the presented findings highlight the potential of benzophenones as a new class of anti-infectives for the treatment of Acanthamoeba keratitis.
proteins
2025-12-16 | Acanthamoeba Spp. Detection in Contact Lens Wearers and Non-Wearers in Iraq
Background: Acanthamoeba spp. are free-living amoebae found in a broad range of environments. Acanthamoeba spp. are responsible for about 20% of keratitis infections in contact lens wearers. We aimed to detect Acanthamoeba spp. and determined the prevalence of Acanthamoeba spp. in contact lens wearers to increase health awareness of the risks of spreading it. Methods: The current study included an investigation of free-living opportunistic amoeba Acanthamoeba spp. in eye swab samples among students in the College of Sciences at the University of Thi-Qar/Iraq. Eighty-eight samples were collected from the eyes of both sexes and for both lens wearers and non-lens wearers from January to April 2024. Results: The current study results showed that the percentage of Acanthamoeba was 4.5% (4/88). All positive cases were in females who were lens wearers only, and no cases were recorded in males or females who had no contact lens. Acanthamoeba spp. were identified morphologically based on characteristics of active trophozoites and cyst forms, and molecular identified by conventional PCR using Acanthamoeba primers JPD1/ JPD2. Additionally, sequencing analysis was performed on positive samples and their alignment sequences were analyzed using BLAST. The results showed a homologous identity (100%) for four samples with the reference isolate Acanthamoeba spp. 18S ribosomal RNA gene )MK390853.1). It was registered as one isolate of Acanthamoeba spp. in gene bank with accession number (PQ661179.1). Conclusion: the current findings indicate the prevalence of Acanthamoeba spp. in contact lens wearers, which is a health risk, especially in females
2025-12-04 | Acanthamoeba Keratitis: Rare but not to be Overlooked
Hornhimnusking af vldum Acanthamoeba (Acanthamoeba keratitis -AK) er afar sjaldgaefur en alvarlegur augnsjkdmur
2025-11-27 | Inhibitory Effect of a Novel Ophthalmic Solution on Acanthamoeba castellanii Adhesion and Biofilm Formation on Human Corneal Epithelium.
Background/Objectives:Acanthamoeba keratitis (AK) is a rare but sight-threatening corneal infection, often associated with contact lens wear and resistant to conventional therapies. Preventive strategies capable of reducing Acanthamoeba adhesion to corneal epithelium may represent an important tool for infection control. This study aimed to evaluate the amebicidal and preventive activity of CORNEIAL MED eye drops against Acanthamoeba castellanii adhesion and early adhesion layer on human corneal epithelium (HCE). Methods: Reconstructed HCE models were exposed to A. castellanii under four experimental conditions: negative control (HCE only), positive control (HCE + A. castellanii), co-incubation with CORNEIAL MED and A. castellanii (Study 1), and treatment with CORNEIAL MED after initial A. castellanii adhesion (Study 2). Adherent amoebae were quantified using EDTA detachment and Neubauer chamber counting. The early adhesion layer was characterized by scanning electron microscopy (SEM). Statistical analysis considered p < 0.05 as significant. Results: In Study 1, simultaneous application of CORNEIAL MED with A. castellanii reduced amoeba adhesion by 33.0 ± 11% compared with controls (p = 0.0529). In Study 2, when the product was applied 3 h after amoeba inoculation, adhesion was significantly reduced by 51.9 ± 6.5% (p < 0.05). SEM confirmed a decrease in amoebic colonization and biofilm density in treated samples. Conclusions: CORNEIAL MED demonstrated a measurable inhibitory effect on A. castellanii adhesion to HCE, particularly when applied after initial pathogen contact. These findings suggest a potential preventive role of CORNEIAL MED in reducing AK risk, although further in vivo studies are warranted.
2025-11-19 | [The fascinating world of free-living amoebae, these remarkably versatile protists].
Free-living amoebae, microorganisms with variable forms, are essential to the ecology. However, some species, such as Naegleria fowleri, Acanthamoeba spp., and Balamuthia mandrillaris, cause severe, though rare, infections including meningitis and keratitis, which are often linked to water. Free-living amoebae are also reservoirs and vectors of various pathogens (bacteria, viruses, and fungi), which facilitates their dissemination and the potential emergence of diseases. Free-living amoebae can also be used as models to study interactions between eukaryotes and prokaryotes. This review explores the ecological, scientific, and pathogenic roles of free-living amoebae and their implications for public health.
2025-07-13 | Complicated Diagnosis and Treatment of Rare Painless Acanthamoeba Keratitis.
Objectives: The aim was to present the complicated diagnostic and therapeutic process of atypical, painless keratitis caused by a cosmopolitan protozoan of the genus Acanthamoeba. Methods: This Case Report describes a medical case involving a 48-year-old woman who occasionally wears soft contact lenses and was referred to our hospital for treatment due to deteriorating visual acuity in her left eye. The diagnostic process included the isolation of amoebae from corneal scrapings and the morphological and molecular identification of the etiological agent of the infection. Results: After examination, painless atypical keratitis was diagnosed, initially considered recurrent herpetic keratitis. However, antiviral treatment did not bring about any improvement. Further observation revealed a dense, central, annular infiltrate on the periphery of the cornea. Despite treatment, the corneal infiltrate did not improve and the patient required therapeutic penetrating keratoplasty. Ultimately, the patient underwent combined surgery: corneal transplantation with cataract phacoemulsification and intraocular lens implantation. The postoperative course was uneventful. Conclusions: Acanthamoeba keratitis should be included in the differential diagnosis of keratitis, even in the absence of its characteristic feature of severe ocular pain, especially in contact lens wearers and patients who have had herpetic keratitis. Infection of the cornea with the Herpes simplex type 1 virus causes nerve degeneration, which probably translates into a painless course of Acanthamoeba castellanii infection.
antibodies
2026-06-29 | In Vitro Inhibition of Bacterial Interactions With Acanthamoeba castellanii by a Monoclonal Antibody Against Galactose-Binding Protein.
Introduction Acanthamoeba castellanii causes amoebic keratitis (AK), primarily in contact lens wearers, and severe lesions may require corneal transplantation. Direct contact between the amoeba and the cornea during AK induction is important for the pathogenic process of the amoeba. From a clinical perspective, identifying molecular factors that regulate this contact-dependent interaction is essential for understanding disease progression and for developing preventive or therapeutic strategies for AK. Methods This study analyzed the effect of treatments with galactose and monoclonal antibodies against the galactose-binding protein (GBP) on the interactions between bacteria and A. castellanii. A. castellanii trophozoites were pre-incubated with monosaccharides (galactose, mannose, or glucose; 100 mM) or monoclonal antibodies against GBP for one hour, followed by co-incubation with pathogenic Escherichia coli O157:H7 (one hour) or non-pathogenic E. coli DH5α (four hours). Bacterial association and invasion were quantified using colony-forming unit (CFU)-based assays following cell lysis, and proteolytic activity was evaluated using gelatin zymography. Statistical analysis was performed using Student's t-test, with P<0.05 considered significant. Results The amoeba-bacteria association after the incubation treatment with mannose was 3.4 times lower than with no treatment. However, the association after the galactose treatment was about 0.7 times lower than with no treatment. In particular, incubation with monoclonal antibodies to GBP showed results very similar to those of mannose. E. coli O157:H7's invasion of the amoebae was reduced by about three times as compared to the association reaction. This reduction in invasion suggests that disruption of lectin-mediated binding may limit the ability of pathogenic bacteria to persist within amoebae. The effects of the incubation treatments with monoclonal antibodies against galactose, mannose, and GBP were so similar that they were almost incomparable. E. coli DH5α's association reaction with galactose was slightly lower than that in the monosaccharide-untreated (no treatment) group. However, it was confirmed that monoclonal antibodies against GBP could reduce the amoeba-bacteria association about 3.65 times more than no treatment. The incubation treatment with antibodies to GBP showed an increase in proteolytic enzyme expression to a degree very similar to that of the mannose treatment group. Conclusion Taken together, these findings indicate that GBP plays a key role in contact-dependent interactions relevant to amoebic pathogenicity. Targeting GBP-mediated pathways may represent a clinically meaningful approach to attenuating host tissue damage and microbial interactions associated with AK.
2025-06-11 | Detection of immunogenic protein components in excretion/secretion products of Acanthamoeba T5 using polyclonal antibodies.
Acanthamoeba is a free-living amoeba widely distributed, responsible for keratitis and granulomatous amoebic encephalitis. The presence of virulence factors in its excretion/secretion products has been demonstrated. Characterisation of these products, including the determination of immunogenic protein components using polyclonal antibodies, could be the basis for the development of new diagnostic tools and help to understand aspects related to its pathogenesis. To identify immunogenic protein components in Acanthamoeba conditioned medium (ACM) and extracellular vesicles (EVs) using polyclonal anti-Acanthamoeba antibodies produced in the laboratory and to evaluate the effect of these antibodies in adhesion and cytopathic effect. Excretion/secretion products were obtained after the axenic culture of a potentially pathogenic environmental Acanthamoeba T5 isolate. The presence of immunogenic components in lysates of trophozoites, ACM and EVs was determined using polyclonal anti-Acanthamoeba antibodies produced in Wistar rats. Proteomic analyses to identify the immunogenic protein components in ACM and EVs were included. Experiments to evaluate the effect of polyclonal anti-Acanthamoeba antibodies in adhesion and cytopathic effect in vitro were also performed in Vero cells. Protein recognition by anti-Acanthamoeba antibodies in lysates, ACM and EVs was demonstrated, and these components were identified using proteomics. Decreases in adhesion and cytopathic effect after the preincubation of trophozoites with antibodies, prior to the contact with cells, were observed. The development of polyclonal antibodies, capable of recognising proteins related to pathogenesis in ACM and EVs, and with significant effects in adhesion, provides an important tool for the search for new therapeutic and diagnostic targets in infections caused by Acanthamoeba.
2025-02-03 | Prevalence of HSV Genomic Signatures Among Acanthamoeba Hosts and Contaminated Lens Cases: A Molecular and Clinical Study.
To document the presence of herpes simplex virus (HSV) genomic signatures among Acanthamoeba hosts recovered from patients with Acanthamoeba keratitis (AK) and in contaminated lens cases. We used a combination of PCR sequencing and shotgun metagenomics to detect and confirm the presence of HSV genomic signatures in Acanthamoeba hosts and lens cases. Clinical outcomes were correlated with the prevalence of host HSV signatures. HSV genomic signatures were detected in 26% (n = 6) of Acanthamoeba hosts recovered from patients with culture confirmed AK. HSV-1 and HSV-2 or both were identified in 33%, 50%, and 17% of isolates, respectively. Fifty-two percent of patients (12/23) were misdiagnosed initially as presenting with HSV keratitis. Patients with HSV-positive Acanthamoeba isolates had a mean best-corrected visual acuity of 1.43 LogMAR at diagnosis and 0.53 LogMAR at follow-up, compared with 1.85 and 0.92 LogMAR, respectively, in HSV-negative cases. Contact lens use was identified as a risk factor in 83% of 18 patients. We detected 46,597 viral signatures in 5 of 14 contaminated lens cases (35.7%). Distribution included 33% bacteriophages, 8.2% giant viruses, 4.1% nonhuman Herpesviridae, and 0.39% human Herpesviridae. Among the 184 human Herpesviridae genomic signatures, HSV types 1 or 2 or both were documented in 5.7%, VZV in 39.7%, HHV7 in 38.6%, HHV6 in 15.0%, and Epstein-Barr virus in 0.5%. This study is the first to identify HSV-positive genomic signatures in clinical AK isolates and/or contact lens cases. Taken together, the high prevalence of HSV genomic signatures in both amebic hosts and lens cases, might signal an unrecognized Acanthamoeba-HSV association and the need for reassessing current management.
2024-02-04 | Epidemiology of and Genetic Factors Associated with Acanthamoeba Keratitis
Acanthamoeba keratitis (AK) is a severe, rare protozoal infection of the cornea. Acanthamoeba can survive in diverse habitats and at extreme temperatures. AK is mostly seen in contact lens wearers whose lenses have become contaminated or who have a history of water exposure, and in those without contact lens wear who have experienced recent eye trauma involving contaminated soil or water. Infection usually results in severe eye pain, photophobia, inflammation, and corneal epithelial defects. The pathophysiology of this infection is multifactorial, including the production of cytotoxic proteases by Acanthamoeba that degrades the corneal epithelial basement membrane and induces the death of ocular surface cells, resulting in degradation of the collagen-rich corneal stroma. AK can be prevented by avoiding risk factors, which includes avoiding water contact, such as swimming or showering in contact lenses, and wearing protective goggles when working on the land. AK is mostly treated with an antimicrobial therapy of biguanides alone or in combination with diaminidines, although the commercial availability of these medicines is variable. Other than anti-amoeba therapies, targeting host immune pathways in Acanthamoeba disease may lead to the development of vaccines or antibody therapeutics which could transform the management of AK.
2021-11-04 | The increasing importance of Vermamoeba vermiformis.
Vermamoeba vermiformis are one of the most prevalent free-living amoebae. These amoebae are ubiquitous and also thermotolerant. Of concern, V. vermiformis have been found in hospital water networks. Furthermore, associations between V. vermiformis and pathogenic bacteria have been reported, such as Legionella pneumophila. Moreover, V. vermiformis are well known to host viruses, bacteria, and other microorganisms and cases of keratitis due to V. vermiformis in conjunction with other amoebae have been reported. Despite the preceding, the medical importance of V. vermiformis is still an ongoing discussion and its genome has been only recently sequenced. Herein, we present a review of the current understanding of the biology and pathogenesis pertaining to V. vermiformis, as well as its' role as an etiological agent and trojan horse. An approach known as theranostics which combines both diagnosis and therapy could be utilized to eradicate and diagnose keratitis cases caused by such amoebae. Given the rise in global warming, it is imperative to investigate these rarely studied amoebae and to understand their importance in human health.
cell therapies
2026-04-01 | Anti-amoebic and biocompatible properties of nephrite-embedded contact lenses.
Acanthamoeba keratitis remains a severe complication of contact lens (CL) wear, and preventing trophozoite adhesion to the lens surface is a critical step in preventing infection. This study investigated the anti-amoebic performance and ocular safety of soft CLs embedded with nephrite, a mineral with inherent antimicrobial activity. Clear, cosmetic, and nephrite-embedded CLs were compared. We uniformly dispersed nephrite powder onto the inner mold surface and encapsulated it within the polymer network using the Flusier method, preventing direct corneal exposure while maintaining transparency. In vitro assays demonstrated that nephrite-embedded CLs had significantly fewer adherent Acanthamoeba lugdunensis trophozoites than clear (p = 0.0093) and cosmetic CLs (p = 0.0068). SEM images showed reduced pseudopodia formation and round, contracted morphology of the attached trophozoites, suggesting early encystment. Although AFM revealed greater surface roughness than clear lenses, Acanthamoeba adhesion was still markedly lower. In vivo evaluation in rabbits showed no corneal opacity, no histologic inflammatory response, and no significant differences in corneal thickness or endothelial cell density compared with other groups. These findings indicate that the nephrite-embedded CLs developed in this study effectively inhibited Acanthamoeba adhesion while maintaining corneal safety, suggesting nephrite as a promising material for CLs with anti-amoebic properties.
2025-07-30 | Achromobacter in the Conjunctival Sac Microbiota: Potential Association With Acanthamoeba Keratitis Related to Orthokeratology Lenses.
Acanthamoeba keratitis (AK) is a severe infection linked to orthokeratology lens use, whereas the involvement of conjunctival microbiota in AK remains poorly understood. This study investigates microbiota dysbiosis in AK pathogenesis to inform microbiota-based interventions. Conjunctival swabs from 14 patients with AK and 10 healthy controls underwent 16S rRNA sequencing. Microbiome analysis compared diversity, taxa, and metabolic pathways. Functional assays quantified Achromobacter-enhanced Acanthamoeba adhesion and migration. Metagenomics and fluorescence in situ hybridization (FISH) with species-specific probes confirmed endosymbiosis. Patients with AK showed reduced bacterial diversity compared with the healthy controls (P < 0.001) but similar richness. Relative abundance of Achromobacter in the AK group was higher compared to the healthy control group (P < 0.001). Achromobacter dominated microbiota among the AK group, being identified as a key biomarker via the linear discriminant analysis effect size (LEfSe). In vitro, Achromobacter increased Acanthamoeba adhesion (P = 0.007) and the migration area (P < 0.05). Metagenomic analysis and FISH further showed Achromobacter spp. as potential endosymbionts of Acanthamoeba. Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed upregulated phenylalanine, fatty acid, and propanoate metabolism in the AK group (all P < 0.001). MetaCyc highlighted enriched pyruvate fermentation to isobutanol, aerobic respiration I, and L-isoleucine biosynthesis II in the AK group (P < 0.001). AK-associated conjunctival dysbiosis features Achromobacter dominance, reduced diversity, and altered metabolism. Achromobacter is associated with enhanced adhesion and migration of Acanthamoeba, indicating a possible symbiotic interaction and its potential as a biomarker and therapeutic target.
2023-09-29 | Amniotic membrane use for resistant infectious and non-infectious corneal ulcer
In corneal ulcers, amniotic membrane (AM) is utilised for supporting damaged tissues, shield and protect deficiencies from further deterioration or breakdown from external influences and encourage re-cellularization of the cornea. Anti-fibrotic, anti-inflammatory, and anti-angiogenic effects, as well as an antimicrobial impact and the encouragement of epithelial and stromal repair are some of the qualities possessed by amniotic membrane. Surgical grafting to the ocular surface, non-surgical patching with a carriage device, amniotic membrane extract eye drops (AMEED), amniotic membrane with umbilical cord blood, and amniotic membrane with placental fluid are certain uses and techniques for utilising AM in corneal ulcers and corneal epithelial deficiencies.
2022-03-11 | Commensals Serve as Natural Barriers to Mammalian Cells during Acanthamoeba castellanii Invasion.
Acanthamoeba castellanii is a free-living, pathogenic ameba found in the soil and water. It invades the body through ulcerated skin, the nasal passages, and eyes and can cause blinding keratitis and granulomatous encephalitis. However, the mechanisms underlying the opportunistic pathogenesis of A. castellanii remain unclear. In this study, we observed that commensal bacteria significantly reduced the cytotoxicity of the ameba on mammalian cells. This effect occurred in the presence of both Gram-positive and Gram-negative commensals. Additionally, commensals mitigated the disruption of cell junctions. Ex vivo experiments on mouse eyeballs further showed that the commensals protected the corneal epithelial layer. Together, these findings indicate that A. castellanii is pathogenic to individuals with a dysbiosis of the microbiota at infection sites, further highlighting the role of commensals as a natural barrier during parasite invasion. IMPORTANCE Acanthamoeba castellanii, an opportunistic protozoan widely present in the environment, can cause Acanthamoeba keratitis and encephalitis in humans. However, only a few reports describe how the ameba acts as an opportunistic pathogen. Our study showed that the normal microbiota interfered with the cytotoxicity of Acanthamoeba, persevered during Acanthamoeba invasion, and reduced corneal epithelium peeling in the mouse eyeball model. This suggests that commensals may act as a natural barrier against Acanthamoeba invasion. In future, individuals who suffer from Acanthamoeba keratitis should be examined for microbiota absence or dysbiosis to reduce the incidence of Acanthamoeba infection in clinical settings.
2020-03-11 | [Necrotizing scleritis after acanthamoeba keratitis].
Nodular scleritis and necrotizing scleritis are rare complications of acanthamoeba keratitis. This article presents the case of a 61-year-old female patient who had suffered from persistent keratitis in the right eye for more than 4 months. The patient was initially treated with propamidine isethionate and polyhexamethylene biguanide eye drops. A penetrating limbo-keratoplasty was performed. Examination of the corneal explant showed acanthamoeba cysts. In the following 5 months the sclera showed recurrent abscesses. A total of two thermal cauterizations and three amniotic membrane grafts were carried out. To our knowledge, this is the first case of sclerokeratitis after acanthamoeba keratitis which was treated with a combination of thermal cauterization and amniotic membrane transplantation. Further studies are necessary to investigate this procedure as an alternative to the established cryotherapy.Die noduläre Skleritis und die nekrotisierende Skleritis sind seltene Komplikationen der Akanthamöbenkeratitis. Wir präsentieren den Fall einer 61-jährigen Patientin, die seit mehr als 4 Monaten an einer persistierenden Keratitis am rechten Auge litt. Es wurde eine perforierende Limbokeratoplastik durchgeführt. Die Untersuchung des Hornhautexplantates zeigte Akanthamöbenzysten. In den folgenden 5 Monaten zeigte die Sklera rezidivierende Abszedierungen. Wir führten insgesamt 2 Thermokauterisationen und 3 Amnionmembrantransplantationen durch. Nach unserem Wissen ist dies der erste Fall einer Sklerokeratitis nach einer Akanthamöbenkeratitis, welcher mit einer Kombination aus Thermokauterisation und Amnionmembrantransplantation behandelt wurde. Weitere Studien sind notwendig, um dieses Verfahren als Alternative zur etablierten Kryotherapie zu untersuchen.
other
2025-11-25 | PREVALENCE OF ACANTHAMOEBA IN MALAYSIAN WATER SOURCES AND ITS PUBLIC HEALTH IMPLICATIONS: A SCOPING REVIEW
Acanthamoeba, a free-living amoeba, poses significant public health risks, causing infections like Acanthamoeba keratitis (AK) in contact lens users and granulomatous amoebic encephalitis (GAE) in immunocompromised individuals. This scoping review examines Acanthamoeba prevalence in Malaysian water sources using a PRISMA-ScR-guided methodology. Literature searches from 2010 to 2024 identified studies focused on natural and artificial water sources, employing microscopy, culture, and molecular techniques for detection and genotyping. The findings revealed high contamination rates: 54.4 % in swimming pools, 90.2 % in drinking water, 100.0 % in recreational rivers, 86.7 % in lakes, 76.0 % in hot springs, and 72.0 % in marine waters, with the pathogenic T4 genotype predominating. Acanthamoeba’s role as a "Trojan horse," harbouring amoeba-resistant bacteria (ARBs), amplifies public health concerns. Prevalence in Malaysia exceeds neighbouring countries, influenced by factors such as climate and water management practices. Key research gaps include the lack of longitudinal studies and insufficient exploration of environmental influences on Acanthamoeba virulence. The review highlights the need for improved water treatment protocols, advanced diagnostics, and targeted public health measures. Effective strategies should prioritize thorough monitoring, public education, and enhanced water safety practices to mitigate risks and protect high-risk populations.
2025-06-26 | Navigating the Challenges of Acanthamoeba Keratitis: Current Trends and Future Directions.
Acanthamoeba keratitis (AK) is a vision-threatening eye infection induced by the free-living species of the amoeba genus Acanthamoeba, presenting considerable therapeutic difficulties due to its frequently delayed diagnosis, chronic nature, and resistance to standard treatments. This review examines the changing landscape of AK, emphasizing recent developments in pathogenesis, diagnosis, and treatment. We examine the molecular pathways that enable Acanthamoeba invasion and persistence in the cornea, and how comprehending these processes can inform the creation of more effective treatment therapies. The review emphasizes current advancements in diagnostic methodologies, such as PCR-based tests and in vivo confocal imaging, which have enhanced early detection rates and diagnostic precision. Furthermore, we analyze contemporary treatment modalities, including antimicrobial therapy and surgical procedures, while recognizing the difficulties presented by antimicrobial resistance and the lack of standardized treatment protocols. This review seeks to deliver a thorough examination of AK, presenting insights into cutting-edge treatments and pinpointing essential areas for future research to address the persistent hurdles in controlling this potentially devastating ocular infection.
2024-03-04 | Could giant viruses be considered as a biotechnological tool for preventing and controlling Acanthamoeba infections?
The aim of the study was to evaluate the efficiency of mimivirus as a potential therapeutic and prophylactic tool against Acanthamoeba castellanii, the etiological agent of Acanthamoeba keratitis, a progressive corneal infection, that is commonly associated with the use of contact lenses and can lead to blindness if not properly treated. Mimivirus particles were tested in different multiplicity of infection, along with commercial multipurpose contact lenses' solutions, aiming to assess their ability to prevent encystment and excystment of A. castellanii. Solutions were evaluated for their amoebicidal potential and cytotoxicity in MDCK cells, as well as their effectiveness in preventing A. castellanii damage in Madin-Darby canine kidney (MDCK) cells. Results indicated that mimivirus was able to inhibit the formation of A. castellanii cysts, even in the presence of Neff encystment solution. Mimivirus also showed greater effectiveness in controlling A. castellanii excystment compared to commercial solutions. Additionally, mimivirus solution was more effective in preventing damage caused by A. castellanii, presented greater amoebicidal activity, and were less cytotoxic to MDCK cells than commercial MPS. Mimivirus demonstrates a greater ability to inhibit A. castellanii encystment and excystment compared to commercial multipurpose contact lens solutions. Additionally, mimivirus is less toxic to MDCK cells than those commercial solutions. New studies utilizing in vivo models will be crucial for confirming safety and efficacy parameters.
2023-12-18 | Epidemiology and Genetic Factors Associated with <em>Acanthamoeba</em> Keratitis
Acanthamoeba keratitis (AK) is a severe, rare protozoal infection of the cornea. Acanthamoeba can survive in diverse habitats and at extreme temperatures. AK is mostly seen in contact lens wearers, whose lenses have become contaminated, or a history of water exposure, and in those without contact lens wear, who have experienced recent eye trauma involving contaminated soil or water. Infection usually results in severe eye pain, photophobia, inflammation, and corneal epithelial defects. The pathophysiology of this infection is multifactorial, including the production of cytotoxic proteases by Acanthamoeba that degrades the corneal epithelial basement membrane and induce the death of ocular surface cells, resulting in degradation of the collagen-rich corneal stroma. AK can be prevented by avoiding the risk factors, which include avoiding water contact such as swimming or showering in contact lenses, and wearing protective goggles when working on the land. AK is mostly treated with an antimicrobial therapy of biguanides alone or in combination with diaminidines, although the commercial availability of these medicines is variable. Other than anti-amoeba therapies, targeting host immune pathways in Acanthamoeba disease may lead to the development of vaccines or antibody therapeutics which could transform the management of AK.
2021-01-12 | Ac-HSP20 Is Associated With the Infectivity and Encystation of Acanthamoeba castellanii
Acanthamoeba castellanii is a pathogenic and opportunistic free-living amoeba that causes Acanthamoeba keratitis (AK) and granulomatous amebic encephalitis (GAE) in immunocompromised individuals. The biological and pathogenic characterizations behind this opportunistic protozoan is not fully understood. This study aimed to determine the biological functions of heat shock protein (HSP)-20 of A. castellanii ( Ac -HSP20) involved in the maintenance of life cycle and the infectivity of A. castellanii . Immunoscreening A. castellanii cDNA library with A. castellanii infected rabbit sera identified three positive clones, one of them was a putative heat shock protein ( Ac -HSP20). The recombinant 23 kDa Ac -HSP20 protein (r Ac -HSP20) was successfully expressed in Escherichia coli BL21 (DE3) and purified using metal affinity chromatography. The rabbits immunized with r Ac -HSP20 produced high titer antibody (1:25,600). Immunolocalization with the antibody identified the expression of native Ac -HSP20 on the surface of both A. castellanii trophozoites and cysts. Further, Western blot with antibody identified that the expression of native Ac-HSP20 was 7.5 times higher in cysts than in trophozoites. Blocking Ac -HSP20 on the membrane of trophozoites with specific antibody or silencing Ac-hsp20 gene transcription by siRNA inhibited their transformation into cysts at the early stage but returned to normal at the late stage by stimulating the transcription of Ac-hsp20 . Incubation of trophozoites with anti- Ac -HSP20 IgG increased macrophage-involved phagocytosis to the protozoa and inhibited trophozoite infectivity on the cornea of rabbits compared with that without antibody. Our study provides that Ac -HSP20 is a surface antigen involved in the encystation and infectivity of A. castellanii and thus an important target for vaccine and drug development.
small molecules
2026-09-03 | Clinical and Microbiological Comparison of Acanthamoeba Keratitis According to the Existence of Hypopyon.
To compare the clinical outcomes of patients with Acanthamoeba keratitis (AK) according to the existence of hypopyon. A retrospective analysis of microbiologically confirmed AK cases over the past 15 years was conducted. Nine eyes with hypopyon (group 1) and 29 eyes without hypopyon (group 2) were analyzed to study the differences in these 2 groups of AK eyes. The cases were diagnosed using conventional microscopic analysis with staining techniques with smear and cultural methods and were treated according to diagnosis. There was no difference in the demographic profiles and duration of symptoms before first visit in the 2 groups. The presenting corneal features, presenting best-corrected visual acuity, and final best-corrected visual acuity were significantly different between groups (P = 0.042, P = 0.002, P = 0.001, respectively). There were significant differences between groups in microbiologic diagnosis (P < 0.001): bacteria with Acanthamoeba (0/9 in group 1 vs. 4/29 in group 2), fungus with Acanthamoeba (7/9 in group 1 vs. 0/29 in group 2), and Acanthamoeba only (2/9 in group 1 vs. 25/29 in group 2). The eyes with fungus and AK coinfection showed worse visual prognosis. AK with hypopyon can be a representative sign of fungal coinfection, which has a trend of a worse visual prognosis. It should immediately alert clinicians to initiate early antifungals and antiamoebics.
2026-09-02 | Anti-Acanthamoeba Activities of Silica Nanoformulated Quercetin Compound Derived From Annona muricata L. Leaves.
Acanthamoeba spp., free-living amoebae (FLA), are the causative agents of Acanthamoeba keratitis (AK) and granulomatous amoebic encephalitis (GAE). These diseases have limited treatment options; therefore, drug discovery remains a critical priority. This study evaluated the leaf extract of Annona muricata L., which contains quercetin, formulated in silica nanoparticles against Acanthamoeba species. The anti-Acanthamoeba effect of the nanoformulation was determined by its MIC and percentage viability. Scanning electron microscopy confirmed the nanocompound-directed effect on trophozoites and cysts. The nanoquercetin exhibited anti-Acanthamoeba activity, showing greater effectiveness against the trophozoites of Acanthamoeba polyphaga ATCC 30461 (AP) (MIC > 2 mg/mL) compared to cysts (MIC > 8 mg/mL) and against A. triangularis WU19001 (AT) trophozoites (MIC > 4 mg/mL) compared to cysts (MIC > 8 mg/mL). It also showed reduced activity against Acanthamoeba castellanii ATCC 30010 (AC3) trophozoites (MIC > 4 mg/mL) compared to cysts (MIC > 8 mg/mL) and A. castellanii ATCC50739 (AC5) trophozoites (MIC > 4 mg/mL) compared to cysts (MIC > 8 mg/mL). The anti-Acanthamoeba activity of A. muricata on all four species was compared to chlorhexidine, which had MIC values of 0.008 mg/mL for AC3 and AT trophozoites and 0.016 mg/mL for AC5 and AP trophozoites. For cysts, chlorhexidine exhibited MIC values of > 0.0128 mg/mL for AC3 and AP, 0.032 mg/mL for AC5, and 0.064 mg/mL for AT. All MIC values demonstrated inhibition of Acanthamoeba species with reduced viability exceeding 90%. SEM findings revealed morphological changes in Acanthamoeba, indicating parasitic inhibition. Quercetin-loaded silica nanoparticles demonstrated anti-Acanthamoeba activity with low cytotoxicity. This indicates the potential of quercetin as a nano-enhanced natural therapeutic or contact lens-care agent. This study is limited by its in vitro design, small parasite strain panel, and potential nanoparticle variability. This highlights the need for in vivo validation, ocular/contact lens model testing, and formulation optimization for improved cysticidal efficacy.
2026-08-28 | An expert's guide to Acanthamoeba keratitis pharmacology.
Acanthamoeba keratitis (AK) is a rare but sight-threatening corneal infection, increasingly associated with contact lens wear, whose treatment remains constrained by the lack of agents that are both cysticidal and well tolerated by the cornea. This review appraises the current pharmacological landscape and the obstacles impeding therapeutic progress. PubMed, Scopus, Google Scholar, and Web of Science were searched for articles published between January 2000 and June 2026 using combinations of the terms 'Acanthamoeba keratitis,' 'treatment,' 'pharmacology,' 'amoebicidal' and individual drug names; reference lists were hand-searched. We provide an overview of the epidemiology, biology, life cycle and pathogenic mechanisms of AK. We then critically evaluate current clinical management strategies and their inherent limitations before providing a comprehensive synthesis of recent in vitro and in vivo investigations to identify effective drugs that target both the cyst and trophozoite stages with low cytotoxicity. Prolonged, intensive topical regimens impose a substantial burden that drives poor adherence and relapse, and standardized protocol-based dosing offers the most immediate practical gain. The field now requires agents combining potent cysticidal activity with corneal biocompatibility, alongside standardized assays and validated ex vivo models, so that promising laboratory activity translates into genuine clinical benefit.
2026-08-27 | Oxidative Stress and Redox Imbalance in Acanthamoeba Keratitis: Host-Parasite Crosstalk and Therapeutic Perspectives.
Acanthamoeba keratitis is a severe, sight-threatening corneal infection caused by free-living amoebae widely distributed in soil, natural waters, and domestic or recreational water systems. Following entry into the corneal microenvironment, disease develops within a redox conflict in which host-derived reactive oxygen species (ROS) and reactive nitrogen species (RNS) may contribute to parasite control while simultaneously promoting inflammation and tissue injury. Acanthamoeba responds through an adaptable defense network involving superoxide dismutases, catalase, thioredoxin- and glutathione-dependent systems, peroxiredoxins, and mitochondrial mechanisms. However, most mechanistic evidence derives from in vitro trophozoite studies, whereas the contribution of individual redox pathways to mature cyst survival and persistence in the infected cornea remains poorly defined. This review critically integrates evidence on host oxidative and nitrosative responses, parasite antioxidant adaptation, bidirectional redox crosstalk, and redox-modulating therapeutic strategies. Current data support neither indiscriminate suppression nor uncontrolled amplification of reactive species. Instead, future interventions should achieve spatially, temporally, and stage-specifically controlled redox modulation that weakens parasite defenses while preserving corneal integrity and essential innate immune functions. Progress will require causal validation of redox targets, separate assessment of trophozoites and mature cysts, long-term excystation and regrowth assays, evaluation across clinical isolates and genotypes, and parallel assessment of host-cell toxicity in clinically relevant corneal models.
2026-08-20 | Benzophenone Derivatives as a New Class of Anti-Infectives Against Acanthamoeba spp.
The rare but sight-threatening ocular disease Acanthamoeba keratitis represents a therapeutic challenge due to its causative agents, Acanthamoeba spp., being able to penetrate the corneal epithelium and persist in the corneal stroma as dormant and highly resistant cysts. To combat the scarcity and limitations of current treatment options, we developed a new class of anti-infective agents against Acanthamoeba spp. based on Ro 48-8071 (3), a known inhibitor of the human oxidosqualene cyclase (hOSC). The compound design was guided by docking studies using a homology model of the cycloartenol synthase (CAS), which is the homolog of hOSC found in Acanthamoeba spp. and essential for the parasite's steroid biosynthesis. Among the synthesized Ro 48-8071 analogues, MSHG19 (4f) evoked the most potent activity against Acanthamoeba hatchetti leading to total eradication of trophozoites and cysts at single-digit micromolar concentrations, whereas having no effects within this concentration range on the viability of human corneal epithelial cells. With our initial set of synthesized compounds, we established a first structure-activity relationship model, which nicely aligns with the predicted binding mode of 4f to CAS. Overall, the presented findings highlight the potential of benzophenones as a new class of anti-infectives for the treatment of Acanthamoeba keratitis.
proteins
2025-12-16 | Acanthamoeba Spp. Detection in Contact Lens Wearers and Non-Wearers in Iraq
Background: Acanthamoeba spp. are free-living amoebae found in a broad range of environments. Acanthamoeba spp. are responsible for about 20% of keratitis infections in contact lens wearers. We aimed to detect Acanthamoeba spp. and determined the prevalence of Acanthamoeba spp. in contact lens wearers to increase health awareness of the risks of spreading it. Methods: The current study included an investigation of free-living opportunistic amoeba Acanthamoeba spp. in eye swab samples among students in the College of Sciences at the University of Thi-Qar/Iraq. Eighty-eight samples were collected from the eyes of both sexes and for both lens wearers and non-lens wearers from January to April 2024. Results: The current study results showed that the percentage of Acanthamoeba was 4.5% (4/88). All positive cases were in females who were lens wearers only, and no cases were recorded in males or females who had no contact lens. Acanthamoeba spp. were identified morphologically based on characteristics of active trophozoites and cyst forms, and molecular identified by conventional PCR using Acanthamoeba primers JPD1/ JPD2. Additionally, sequencing analysis was performed on positive samples and their alignment sequences were analyzed using BLAST. The results showed a homologous identity (100%) for four samples with the reference isolate Acanthamoeba spp. 18S ribosomal RNA gene )MK390853.1). It was registered as one isolate of Acanthamoeba spp. in gene bank with accession number (PQ661179.1). Conclusion: the current findings indicate the prevalence of Acanthamoeba spp. in contact lens wearers, which is a health risk, especially in females
2025-12-04 | Acanthamoeba Keratitis: Rare but not to be Overlooked
Hornhimnusking af vldum Acanthamoeba (Acanthamoeba keratitis -AK) er afar sjaldgaefur en alvarlegur augnsjkdmur
2025-11-27 | Inhibitory Effect of a Novel Ophthalmic Solution on Acanthamoeba castellanii Adhesion and Biofilm Formation on Human Corneal Epithelium.
Background/Objectives:Acanthamoeba keratitis (AK) is a rare but sight-threatening corneal infection, often associated with contact lens wear and resistant to conventional therapies. Preventive strategies capable of reducing Acanthamoeba adhesion to corneal epithelium may represent an important tool for infection control. This study aimed to evaluate the amebicidal and preventive activity of CORNEIAL MED eye drops against Acanthamoeba castellanii adhesion and early adhesion layer on human corneal epithelium (HCE). Methods: Reconstructed HCE models were exposed to A. castellanii under four experimental conditions: negative control (HCE only), positive control (HCE + A. castellanii), co-incubation with CORNEIAL MED and A. castellanii (Study 1), and treatment with CORNEIAL MED after initial A. castellanii adhesion (Study 2). Adherent amoebae were quantified using EDTA detachment and Neubauer chamber counting. The early adhesion layer was characterized by scanning electron microscopy (SEM). Statistical analysis considered p < 0.05 as significant. Results: In Study 1, simultaneous application of CORNEIAL MED with A. castellanii reduced amoeba adhesion by 33.0 ± 11% compared with controls (p = 0.0529). In Study 2, when the product was applied 3 h after amoeba inoculation, adhesion was significantly reduced by 51.9 ± 6.5% (p < 0.05). SEM confirmed a decrease in amoebic colonization and biofilm density in treated samples. Conclusions: CORNEIAL MED demonstrated a measurable inhibitory effect on A. castellanii adhesion to HCE, particularly when applied after initial pathogen contact. These findings suggest a potential preventive role of CORNEIAL MED in reducing AK risk, although further in vivo studies are warranted.
2025-11-19 | [The fascinating world of free-living amoebae, these remarkably versatile protists].
Free-living amoebae, microorganisms with variable forms, are essential to the ecology. However, some species, such as Naegleria fowleri, Acanthamoeba spp., and Balamuthia mandrillaris, cause severe, though rare, infections including meningitis and keratitis, which are often linked to water. Free-living amoebae are also reservoirs and vectors of various pathogens (bacteria, viruses, and fungi), which facilitates their dissemination and the potential emergence of diseases. Free-living amoebae can also be used as models to study interactions between eukaryotes and prokaryotes. This review explores the ecological, scientific, and pathogenic roles of free-living amoebae and their implications for public health.
2025-07-13 | Complicated Diagnosis and Treatment of Rare Painless Acanthamoeba Keratitis.
Objectives: The aim was to present the complicated diagnostic and therapeutic process of atypical, painless keratitis caused by a cosmopolitan protozoan of the genus Acanthamoeba. Methods: This Case Report describes a medical case involving a 48-year-old woman who occasionally wears soft contact lenses and was referred to our hospital for treatment due to deteriorating visual acuity in her left eye. The diagnostic process included the isolation of amoebae from corneal scrapings and the morphological and molecular identification of the etiological agent of the infection. Results: After examination, painless atypical keratitis was diagnosed, initially considered recurrent herpetic keratitis. However, antiviral treatment did not bring about any improvement. Further observation revealed a dense, central, annular infiltrate on the periphery of the cornea. Despite treatment, the corneal infiltrate did not improve and the patient required therapeutic penetrating keratoplasty. Ultimately, the patient underwent combined surgery: corneal transplantation with cataract phacoemulsification and intraocular lens implantation. The postoperative course was uneventful. Conclusions: Acanthamoeba keratitis should be included in the differential diagnosis of keratitis, even in the absence of its characteristic feature of severe ocular pain, especially in contact lens wearers and patients who have had herpetic keratitis. Infection of the cornea with the Herpes simplex type 1 virus causes nerve degeneration, which probably translates into a painless course of Acanthamoeba castellanii infection.
antibodies
2026-06-29 | In Vitro Inhibition of Bacterial Interactions With Acanthamoeba castellanii by a Monoclonal Antibody Against Galactose-Binding Protein.
Introduction Acanthamoeba castellanii causes amoebic keratitis (AK), primarily in contact lens wearers, and severe lesions may require corneal transplantation. Direct contact between the amoeba and the cornea during AK induction is important for the pathogenic process of the amoeba. From a clinical perspective, identifying molecular factors that regulate this contact-dependent interaction is essential for understanding disease progression and for developing preventive or therapeutic strategies for AK. Methods This study analyzed the effect of treatments with galactose and monoclonal antibodies against the galactose-binding protein (GBP) on the interactions between bacteria and A. castellanii. A. castellanii trophozoites were pre-incubated with monosaccharides (galactose, mannose, or glucose; 100 mM) or monoclonal antibodies against GBP for one hour, followed by co-incubation with pathogenic Escherichia coli O157:H7 (one hour) or non-pathogenic E. coli DH5α (four hours). Bacterial association and invasion were quantified using colony-forming unit (CFU)-based assays following cell lysis, and proteolytic activity was evaluated using gelatin zymography. Statistical analysis was performed using Student's t-test, with P<0.05 considered significant. Results The amoeba-bacteria association after the incubation treatment with mannose was 3.4 times lower than with no treatment. However, the association after the galactose treatment was about 0.7 times lower than with no treatment. In particular, incubation with monoclonal antibodies to GBP showed results very similar to those of mannose. E. coli O157:H7's invasion of the amoebae was reduced by about three times as compared to the association reaction. This reduction in invasion suggests that disruption of lectin-mediated binding may limit the ability of pathogenic bacteria to persist within amoebae. The effects of the incubation treatments with monoclonal antibodies against galactose, mannose, and GBP were so similar that they were almost incomparable. E. coli DH5α's association reaction with galactose was slightly lower than that in the monosaccharide-untreated (no treatment) group. However, it was confirmed that monoclonal antibodies against GBP could reduce the amoeba-bacteria association about 3.65 times more than no treatment. The incubation treatment with antibodies to GBP showed an increase in proteolytic enzyme expression to a degree very similar to that of the mannose treatment group. Conclusion Taken together, these findings indicate that GBP plays a key role in contact-dependent interactions relevant to amoebic pathogenicity. Targeting GBP-mediated pathways may represent a clinically meaningful approach to attenuating host tissue damage and microbial interactions associated with AK.
2025-06-11 | Detection of immunogenic protein components in excretion/secretion products of Acanthamoeba T5 using polyclonal antibodies.
Acanthamoeba is a free-living amoeba widely distributed, responsible for keratitis and granulomatous amoebic encephalitis. The presence of virulence factors in its excretion/secretion products has been demonstrated. Characterisation of these products, including the determination of immunogenic protein components using polyclonal antibodies, could be the basis for the development of new diagnostic tools and help to understand aspects related to its pathogenesis. To identify immunogenic protein components in Acanthamoeba conditioned medium (ACM) and extracellular vesicles (EVs) using polyclonal anti-Acanthamoeba antibodies produced in the laboratory and to evaluate the effect of these antibodies in adhesion and cytopathic effect. Excretion/secretion products were obtained after the axenic culture of a potentially pathogenic environmental Acanthamoeba T5 isolate. The presence of immunogenic components in lysates of trophozoites, ACM and EVs was determined using polyclonal anti-Acanthamoeba antibodies produced in Wistar rats. Proteomic analyses to identify the immunogenic protein components in ACM and EVs were included. Experiments to evaluate the effect of polyclonal anti-Acanthamoeba antibodies in adhesion and cytopathic effect in vitro were also performed in Vero cells. Protein recognition by anti-Acanthamoeba antibodies in lysates, ACM and EVs was demonstrated, and these components were identified using proteomics. Decreases in adhesion and cytopathic effect after the preincubation of trophozoites with antibodies, prior to the contact with cells, were observed. The development of polyclonal antibodies, capable of recognising proteins related to pathogenesis in ACM and EVs, and with significant effects in adhesion, provides an important tool for the search for new therapeutic and diagnostic targets in infections caused by Acanthamoeba.
2025-02-03 | Prevalence of HSV Genomic Signatures Among Acanthamoeba Hosts and Contaminated Lens Cases: A Molecular and Clinical Study.
To document the presence of herpes simplex virus (HSV) genomic signatures among Acanthamoeba hosts recovered from patients with Acanthamoeba keratitis (AK) and in contaminated lens cases. We used a combination of PCR sequencing and shotgun metagenomics to detect and confirm the presence of HSV genomic signatures in Acanthamoeba hosts and lens cases. Clinical outcomes were correlated with the prevalence of host HSV signatures. HSV genomic signatures were detected in 26% (n = 6) of Acanthamoeba hosts recovered from patients with culture confirmed AK. HSV-1 and HSV-2 or both were identified in 33%, 50%, and 17% of isolates, respectively. Fifty-two percent of patients (12/23) were misdiagnosed initially as presenting with HSV keratitis. Patients with HSV-positive Acanthamoeba isolates had a mean best-corrected visual acuity of 1.43 LogMAR at diagnosis and 0.53 LogMAR at follow-up, compared with 1.85 and 0.92 LogMAR, respectively, in HSV-negative cases. Contact lens use was identified as a risk factor in 83% of 18 patients. We detected 46,597 viral signatures in 5 of 14 contaminated lens cases (35.7%). Distribution included 33% bacteriophages, 8.2% giant viruses, 4.1% nonhuman Herpesviridae, and 0.39% human Herpesviridae. Among the 184 human Herpesviridae genomic signatures, HSV types 1 or 2 or both were documented in 5.7%, VZV in 39.7%, HHV7 in 38.6%, HHV6 in 15.0%, and Epstein-Barr virus in 0.5%. This study is the first to identify HSV-positive genomic signatures in clinical AK isolates and/or contact lens cases. Taken together, the high prevalence of HSV genomic signatures in both amebic hosts and lens cases, might signal an unrecognized Acanthamoeba-HSV association and the need for reassessing current management.
2024-02-04 | Epidemiology of and Genetic Factors Associated with Acanthamoeba Keratitis
Acanthamoeba keratitis (AK) is a severe, rare protozoal infection of the cornea. Acanthamoeba can survive in diverse habitats and at extreme temperatures. AK is mostly seen in contact lens wearers whose lenses have become contaminated or who have a history of water exposure, and in those without contact lens wear who have experienced recent eye trauma involving contaminated soil or water. Infection usually results in severe eye pain, photophobia, inflammation, and corneal epithelial defects. The pathophysiology of this infection is multifactorial, including the production of cytotoxic proteases by Acanthamoeba that degrades the corneal epithelial basement membrane and induces the death of ocular surface cells, resulting in degradation of the collagen-rich corneal stroma. AK can be prevented by avoiding risk factors, which includes avoiding water contact, such as swimming or showering in contact lenses, and wearing protective goggles when working on the land. AK is mostly treated with an antimicrobial therapy of biguanides alone or in combination with diaminidines, although the commercial availability of these medicines is variable. Other than anti-amoeba therapies, targeting host immune pathways in Acanthamoeba disease may lead to the development of vaccines or antibody therapeutics which could transform the management of AK.
2021-11-04 | The increasing importance of Vermamoeba vermiformis.
Vermamoeba vermiformis are one of the most prevalent free-living amoebae. These amoebae are ubiquitous and also thermotolerant. Of concern, V. vermiformis have been found in hospital water networks. Furthermore, associations between V. vermiformis and pathogenic bacteria have been reported, such as Legionella pneumophila. Moreover, V. vermiformis are well known to host viruses, bacteria, and other microorganisms and cases of keratitis due to V. vermiformis in conjunction with other amoebae have been reported. Despite the preceding, the medical importance of V. vermiformis is still an ongoing discussion and its genome has been only recently sequenced. Herein, we present a review of the current understanding of the biology and pathogenesis pertaining to V. vermiformis, as well as its' role as an etiological agent and trojan horse. An approach known as theranostics which combines both diagnosis and therapy could be utilized to eradicate and diagnose keratitis cases caused by such amoebae. Given the rise in global warming, it is imperative to investigate these rarely studied amoebae and to understand their importance in human health.
cell therapies
2026-04-01 | Anti-amoebic and biocompatible properties of nephrite-embedded contact lenses.
Acanthamoeba keratitis remains a severe complication of contact lens (CL) wear, and preventing trophozoite adhesion to the lens surface is a critical step in preventing infection. This study investigated the anti-amoebic performance and ocular safety of soft CLs embedded with nephrite, a mineral with inherent antimicrobial activity. Clear, cosmetic, and nephrite-embedded CLs were compared. We uniformly dispersed nephrite powder onto the inner mold surface and encapsulated it within the polymer network using the Flusier method, preventing direct corneal exposure while maintaining transparency. In vitro assays demonstrated that nephrite-embedded CLs had significantly fewer adherent Acanthamoeba lugdunensis trophozoites than clear (p = 0.0093) and cosmetic CLs (p = 0.0068). SEM images showed reduced pseudopodia formation and round, contracted morphology of the attached trophozoites, suggesting early encystment. Although AFM revealed greater surface roughness than clear lenses, Acanthamoeba adhesion was still markedly lower. In vivo evaluation in rabbits showed no corneal opacity, no histologic inflammatory response, and no significant differences in corneal thickness or endothelial cell density compared with other groups. These findings indicate that the nephrite-embedded CLs developed in this study effectively inhibited Acanthamoeba adhesion while maintaining corneal safety, suggesting nephrite as a promising material for CLs with anti-amoebic properties.
2025-07-30 | Achromobacter in the Conjunctival Sac Microbiota: Potential Association With Acanthamoeba Keratitis Related to Orthokeratology Lenses.
Acanthamoeba keratitis (AK) is a severe infection linked to orthokeratology lens use, whereas the involvement of conjunctival microbiota in AK remains poorly understood. This study investigates microbiota dysbiosis in AK pathogenesis to inform microbiota-based interventions. Conjunctival swabs from 14 patients with AK and 10 healthy controls underwent 16S rRNA sequencing. Microbiome analysis compared diversity, taxa, and metabolic pathways. Functional assays quantified Achromobacter-enhanced Acanthamoeba adhesion and migration. Metagenomics and fluorescence in situ hybridization (FISH) with species-specific probes confirmed endosymbiosis. Patients with AK showed reduced bacterial diversity compared with the healthy controls (P < 0.001) but similar richness. Relative abundance of Achromobacter in the AK group was higher compared to the healthy control group (P < 0.001). Achromobacter dominated microbiota among the AK group, being identified as a key biomarker via the linear discriminant analysis effect size (LEfSe). In vitro, Achromobacter increased Acanthamoeba adhesion (P = 0.007) and the migration area (P < 0.05). Metagenomic analysis and FISH further showed Achromobacter spp. as potential endosymbionts of Acanthamoeba. Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed upregulated phenylalanine, fatty acid, and propanoate metabolism in the AK group (all P < 0.001). MetaCyc highlighted enriched pyruvate fermentation to isobutanol, aerobic respiration I, and L-isoleucine biosynthesis II in the AK group (P < 0.001). AK-associated conjunctival dysbiosis features Achromobacter dominance, reduced diversity, and altered metabolism. Achromobacter is associated with enhanced adhesion and migration of Acanthamoeba, indicating a possible symbiotic interaction and its potential as a biomarker and therapeutic target.
2023-09-29 | Amniotic membrane use for resistant infectious and non-infectious corneal ulcer
In corneal ulcers, amniotic membrane (AM) is utilised for supporting damaged tissues, shield and protect deficiencies from further deterioration or breakdown from external influences and encourage re-cellularization of the cornea. Anti-fibrotic, anti-inflammatory, and anti-angiogenic effects, as well as an antimicrobial impact and the encouragement of epithelial and stromal repair are some of the qualities possessed by amniotic membrane. Surgical grafting to the ocular surface, non-surgical patching with a carriage device, amniotic membrane extract eye drops (AMEED), amniotic membrane with umbilical cord blood, and amniotic membrane with placental fluid are certain uses and techniques for utilising AM in corneal ulcers and corneal epithelial deficiencies.
2022-03-11 | Commensals Serve as Natural Barriers to Mammalian Cells during Acanthamoeba castellanii Invasion.
Acanthamoeba castellanii is a free-living, pathogenic ameba found in the soil and water. It invades the body through ulcerated skin, the nasal passages, and eyes and can cause blinding keratitis and granulomatous encephalitis. However, the mechanisms underlying the opportunistic pathogenesis of A. castellanii remain unclear. In this study, we observed that commensal bacteria significantly reduced the cytotoxicity of the ameba on mammalian cells. This effect occurred in the presence of both Gram-positive and Gram-negative commensals. Additionally, commensals mitigated the disruption of cell junctions. Ex vivo experiments on mouse eyeballs further showed that the commensals protected the corneal epithelial layer. Together, these findings indicate that A. castellanii is pathogenic to individuals with a dysbiosis of the microbiota at infection sites, further highlighting the role of commensals as a natural barrier during parasite invasion. IMPORTANCE Acanthamoeba castellanii, an opportunistic protozoan widely present in the environment, can cause Acanthamoeba keratitis and encephalitis in humans. However, only a few reports describe how the ameba acts as an opportunistic pathogen. Our study showed that the normal microbiota interfered with the cytotoxicity of Acanthamoeba, persevered during Acanthamoeba invasion, and reduced corneal epithelium peeling in the mouse eyeball model. This suggests that commensals may act as a natural barrier against Acanthamoeba invasion. In future, individuals who suffer from Acanthamoeba keratitis should be examined for microbiota absence or dysbiosis to reduce the incidence of Acanthamoeba infection in clinical settings.
2020-03-11 | [Necrotizing scleritis after acanthamoeba keratitis].
Nodular scleritis and necrotizing scleritis are rare complications of acanthamoeba keratitis. This article presents the case of a 61-year-old female patient who had suffered from persistent keratitis in the right eye for more than 4 months. The patient was initially treated with propamidine isethionate and polyhexamethylene biguanide eye drops. A penetrating limbo-keratoplasty was performed. Examination of the corneal explant showed acanthamoeba cysts. In the following 5 months the sclera showed recurrent abscesses. A total of two thermal cauterizations and three amniotic membrane grafts were carried out. To our knowledge, this is the first case of sclerokeratitis after acanthamoeba keratitis which was treated with a combination of thermal cauterization and amniotic membrane transplantation. Further studies are necessary to investigate this procedure as an alternative to the established cryotherapy.Die noduläre Skleritis und die nekrotisierende Skleritis sind seltene Komplikationen der Akanthamöbenkeratitis. Wir präsentieren den Fall einer 61-jährigen Patientin, die seit mehr als 4 Monaten an einer persistierenden Keratitis am rechten Auge litt. Es wurde eine perforierende Limbokeratoplastik durchgeführt. Die Untersuchung des Hornhautexplantates zeigte Akanthamöbenzysten. In den folgenden 5 Monaten zeigte die Sklera rezidivierende Abszedierungen. Wir führten insgesamt 2 Thermokauterisationen und 3 Amnionmembrantransplantationen durch. Nach unserem Wissen ist dies der erste Fall einer Sklerokeratitis nach einer Akanthamöbenkeratitis, welcher mit einer Kombination aus Thermokauterisation und Amnionmembrantransplantation behandelt wurde. Weitere Studien sind notwendig, um dieses Verfahren als Alternative zur etablierten Kryotherapie zu untersuchen.
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2025-11-25 | PREVALENCE OF ACANTHAMOEBA IN MALAYSIAN WATER SOURCES AND ITS PUBLIC HEALTH IMPLICATIONS: A SCOPING REVIEW
Acanthamoeba, a free-living amoeba, poses significant public health risks, causing infections like Acanthamoeba keratitis (AK) in contact lens users and granulomatous amoebic encephalitis (GAE) in immunocompromised individuals. This scoping review examines Acanthamoeba prevalence in Malaysian water sources using a PRISMA-ScR-guided methodology. Literature searches from 2010 to 2024 identified studies focused on natural and artificial water sources, employing microscopy, culture, and molecular techniques for detection and genotyping. The findings revealed high contamination rates: 54.4 % in swimming pools, 90.2 % in drinking water, 100.0 % in recreational rivers, 86.7 % in lakes, 76.0 % in hot springs, and 72.0 % in marine waters, with the pathogenic T4 genotype predominating. Acanthamoeba’s role as a "Trojan horse," harbouring amoeba-resistant bacteria (ARBs), amplifies public health concerns. Prevalence in Malaysia exceeds neighbouring countries, influenced by factors such as climate and water management practices. Key research gaps include the lack of longitudinal studies and insufficient exploration of environmental influences on Acanthamoeba virulence. The review highlights the need for improved water treatment protocols, advanced diagnostics, and targeted public health measures. Effective strategies should prioritize thorough monitoring, public education, and enhanced water safety practices to mitigate risks and protect high-risk populations.
2025-06-26 | Navigating the Challenges of Acanthamoeba Keratitis: Current Trends and Future Directions.
Acanthamoeba keratitis (AK) is a vision-threatening eye infection induced by the free-living species of the amoeba genus Acanthamoeba, presenting considerable therapeutic difficulties due to its frequently delayed diagnosis, chronic nature, and resistance to standard treatments. This review examines the changing landscape of AK, emphasizing recent developments in pathogenesis, diagnosis, and treatment. We examine the molecular pathways that enable Acanthamoeba invasion and persistence in the cornea, and how comprehending these processes can inform the creation of more effective treatment therapies. The review emphasizes current advancements in diagnostic methodologies, such as PCR-based tests and in vivo confocal imaging, which have enhanced early detection rates and diagnostic precision. Furthermore, we analyze contemporary treatment modalities, including antimicrobial therapy and surgical procedures, while recognizing the difficulties presented by antimicrobial resistance and the lack of standardized treatment protocols. This review seeks to deliver a thorough examination of AK, presenting insights into cutting-edge treatments and pinpointing essential areas for future research to address the persistent hurdles in controlling this potentially devastating ocular infection.
2024-03-04 | Could giant viruses be considered as a biotechnological tool for preventing and controlling Acanthamoeba infections?
The aim of the study was to evaluate the efficiency of mimivirus as a potential therapeutic and prophylactic tool against Acanthamoeba castellanii, the etiological agent of Acanthamoeba keratitis, a progressive corneal infection, that is commonly associated with the use of contact lenses and can lead to blindness if not properly treated. Mimivirus particles were tested in different multiplicity of infection, along with commercial multipurpose contact lenses' solutions, aiming to assess their ability to prevent encystment and excystment of A. castellanii. Solutions were evaluated for their amoebicidal potential and cytotoxicity in MDCK cells, as well as their effectiveness in preventing A. castellanii damage in Madin-Darby canine kidney (MDCK) cells. Results indicated that mimivirus was able to inhibit the formation of A. castellanii cysts, even in the presence of Neff encystment solution. Mimivirus also showed greater effectiveness in controlling A. castellanii excystment compared to commercial solutions. Additionally, mimivirus solution was more effective in preventing damage caused by A. castellanii, presented greater amoebicidal activity, and were less cytotoxic to MDCK cells than commercial MPS. Mimivirus demonstrates a greater ability to inhibit A. castellanii encystment and excystment compared to commercial multipurpose contact lens solutions. Additionally, mimivirus is less toxic to MDCK cells than those commercial solutions. New studies utilizing in vivo models will be crucial for confirming safety and efficacy parameters.
2023-12-18 | Epidemiology and Genetic Factors Associated with <em>Acanthamoeba</em> Keratitis
Acanthamoeba keratitis (AK) is a severe, rare protozoal infection of the cornea. Acanthamoeba can survive in diverse habitats and at extreme temperatures. AK is mostly seen in contact lens wearers, whose lenses have become contaminated, or a history of water exposure, and in those without contact lens wear, who have experienced recent eye trauma involving contaminated soil or water. Infection usually results in severe eye pain, photophobia, inflammation, and corneal epithelial defects. The pathophysiology of this infection is multifactorial, including the production of cytotoxic proteases by Acanthamoeba that degrades the corneal epithelial basement membrane and induce the death of ocular surface cells, resulting in degradation of the collagen-rich corneal stroma. AK can be prevented by avoiding the risk factors, which include avoiding water contact such as swimming or showering in contact lenses, and wearing protective goggles when working on the land. AK is mostly treated with an antimicrobial therapy of biguanides alone or in combination with diaminidines, although the commercial availability of these medicines is variable. Other than anti-amoeba therapies, targeting host immune pathways in Acanthamoeba disease may lead to the development of vaccines or antibody therapeutics which could transform the management of AK.
2021-01-12 | Ac-HSP20 Is Associated With the Infectivity and Encystation of Acanthamoeba castellanii
Acanthamoeba castellanii is a pathogenic and opportunistic free-living amoeba that causes Acanthamoeba keratitis (AK) and granulomatous amebic encephalitis (GAE) in immunocompromised individuals. The biological and pathogenic characterizations behind this opportunistic protozoan is not fully understood. This study aimed to determine the biological functions of heat shock protein (HSP)-20 of A. castellanii ( Ac -HSP20) involved in the maintenance of life cycle and the infectivity of A. castellanii . Immunoscreening A. castellanii cDNA library with A. castellanii infected rabbit sera identified three positive clones, one of them was a putative heat shock protein ( Ac -HSP20). The recombinant 23 kDa Ac -HSP20 protein (r Ac -HSP20) was successfully expressed in Escherichia coli BL21 (DE3) and purified using metal affinity chromatography. The rabbits immunized with r Ac -HSP20 produced high titer antibody (1:25,600). Immunolocalization with the antibody identified the expression of native Ac -HSP20 on the surface of both A. castellanii trophozoites and cysts. Further, Western blot with antibody identified that the expression of native Ac-HSP20 was 7.5 times higher in cysts than in trophozoites. Blocking Ac -HSP20 on the membrane of trophozoites with specific antibody or silencing Ac-hsp20 gene transcription by siRNA inhibited their transformation into cysts at the early stage but returned to normal at the late stage by stimulating the transcription of Ac-hsp20 . Incubation of trophozoites with anti- Ac -HSP20 IgG increased macrophage-involved phagocytosis to the protozoa and inhibited trophozoite infectivity on the cornea of rabbits compared with that without antibody. Our study provides that Ac -HSP20 is a surface antigen involved in the encystation and infectivity of A. castellanii and thus an important target for vaccine and drug development.
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Drug Discovery Landscape
6 orphan drug designations for Amoebic keratitis, including 1 approved therapy.
6 orphan drug designations for Amoebic keratitis, including 1 approved therapy.
Drug | Therapy type | Orphan designation | Clinical trial status | Sponsor |
|---|---|---|---|---|
polyhexanide | small molecules | FDA | Società Industria Farmaceutica Italiana S.p.A. | |
povidone-iodine 0.5% (w/w) in dimethylsulfoxide 44% (w/w) | small molecules | FDA | Veloce BioPharma, LLC | |
miltefosine | small molecules | FDA | — | Profounda, Inc. |
Polihexanide [AKANTIOR] | small molecules | EMA | Approved | S.I.F.I. Società Industria Farmaceutica Italiana S.p.A. |
Miltefosine | small molecules | EMA | — | Orpha-Devel Handels und Vertriebs GmbH |
Propamidine isethionate 0.1% ophthalmic solution | small molecules | FDA | Bausch & Lomb |
“Terminated” – programs with discontinued clinical trials, or with no updates reported for 5 years after completion or 12 years after the last trial started.
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