Determination of probability of causative pathogen in infectious keratitis using deep learning algorithm of slit-lamp images.

Corneal opacities are important causes of blindness, and their major etiology is infectious keratitis. Slit-lamp examinations are commonly used to determine the causative pathogen; however, their diagnostic accuracy is low even for experienced ophthalmologists. To characterize the "face" of an infected cornea, we have adapted a deep learning architecture used for facial recognition and applied it to determine a probability score for a specific pathogen causing keratitis. To record the diverse features and mitigate the uncertainty, batches of probability scores of 4 serial images taken from many angles or fluorescence staining were learned for score and decision level fusion using a gradient boosting decision tree. A total of 4306 slit-lamp images including 312 images obtained by internet publications on keratitis by bacteria, fungi, acanthamoeba, and herpes simplex virus (HSV) were studied. The created algorithm had a high overall accuracy of diagnosis, e.g., the accuracy/area under the curve for acanthamoeba was 97.9%/0.995, bacteria was 90.7%/0.963, fungi was 95.0%/0.975, and HSV was 92.3%/0.946, by group K-fold validation, and it was robust to even the low resolution web images. We suggest that our hybrid deep learning-based algorithm be used as a simple and accurate method for computer-assisted diagnosis of infectious keratitis.

Diagnostic approach and epidemiology of Microbial Keratitis: findings from an Italian Tertiary Care center.

Rapid identification of causative microorganisms of microbial keratitis (MK) and knowledge of the most common local pathogens are prerequisites for rational antimicrobial therapy. We retrospectively reviewed the characteristics of MK diagnosed at the IRCCS Arcispedale Santa Maria Nuova of Reggio Emilia (Italy) in a 5-years period, where the Ophthalmologist Unit is a reference center for corneal infections. During the study period, 183 MK were evaluated through corneal scrapings cultures. The positivity rate was 54,1%. A total of 107 microorganisms have been isolated: Acanthamoeba species was the etiologic agent in 19 cases. Pseudomonas aeruginosa and Staphylococcus aureus were more frequently isolated in bacterial keratitis, while Fusarium spp., Candida albicans, and Alternaria alternata were predominant among the fungal isolates. Strict cooperation between ophthalmologists and clinical microbiologists is advisable to allow the best diagnostic approach for MK.

Detection of free-living amoebae in domestic cats with and without naturally-acquired keratitis.

Pathogenic free-living amoebae, most notably Acanthamoeba spp., are important pathogens of the human cornea. The importance of infection with free-living amoebae in cats with keratitis is currently unclear. The aim of this study was to determine the frequency of amoeba detection in corneas of cats with naturally-acquired keratitis and in the ocular surface microflora of cats without ocular disease. Clinical ophthalmic and in vivo corneal confocal microscopic examinations were performed on 60 cats with keratitis. Corneal scrapings were analyzed by amoeba culture; cytological evaluation; and Acanthamoeba, Hartmannella, and Vahlkampfia PCR assays. Following ophthalmic examination, conjunctival specimens collected from 60 cats without clinically apparent ocular disease were analyzed similarly. In one cat with ulcerative keratitis, amoeba cysts and trophozoites were detected by in vivo corneal confocal microscopy; an Acanthamoeba sp. was isolated from corneal specimens and detected by Acanthamoeba PCR assay; and suppurative corneal inflammation was present cytologically. An Acanthamoeba sp. was isolated from conjunctival specimens from one cat without clinically apparent ocular disease, but with suppurative inflammation demonstrated cytologically. Both Acanthamoeba isolates belonged to the T4 genotype. Naegleria-like amoebae were isolated in samples from two cats with keratitis and seven cats without clinical ocular disease, but amoebae were not detected by the other assays in these samples. Amoeba detection by culture was significantly (P = 0.01) associated with cytologically diagnosed corneoconjunctival inflammation. This study identified naturally-acquired Acanthamoeba keratitis in cats. Detection of Naegleria-like amoebae in samples from cats with and without keratitis is of uncertain pathological significance.

Improved Outcome of Pythium Keratitis With a Combined Triple Drug Regimen of Linezolid and Azithromycin.

PURPOSE: To describe the clinical features, microbiological profile, and outcome of a series of cases of Pythium keratitis treated with topical and oral linezolid and topical azithromycin eye drops. METHODS: This was a retrospective interventional case series of microbiologically and/or histopathologically proven cases of Pythium keratitis seen between October 2016 and December 2019. All patients received a combination of topical linezolid and/or azithromycin eye drops with oral linezolid. Analysis of demographic data, predisposing risk factors, microbiological results, treatment regimen, visual acuity, surgical intervention, and final outcome was performed. A subgroup analysis of cases >6 mm in size was performed. Success was defined as complete resolution on medical management. Failure was defined as worsening of infection necessitating therapeutic penetrating keratoplasty or evisceration. RESULTS: Of 21 cases, 2 were lost to follow up, 1 was diagnosed on histopathology, and 1 received only topical linezolid. Characteristic microbiological features were noted on 10% potassium hydroxide calcofluor white wet mount in 20/21 (95.23%) and Gram stain in 18/21 (85.71%). On triple drug regimen, 14/17 cases (82.35%) resolved. Average time to resolution was 87.64 ± 44.44 days. More than 60% infiltrates (13/21) were large, and 66.66% infiltrates resolved in 109.3 ± 57.06 days. Of the 5 failures, 4 needed therapeutic keratoplasty and 1 needed evisceration. All grafts failed. CONCLUSIONS: The dual topical drug regimen with oral linezolid has good cure rates (over 80%) for Pythium keratitis over prolonged duration. It is recommended to persevere with medical therapy even in large infiltrates because more than two thirds resolved.

Linezolid shows high safety and efficacy in the treatment of Pythium insidiosum keratitis in a rabbit model.

Efficacy and safety of three antibiotics (Linezolid-LZ, 0.2%; Azithromycin-AZ, 1%; Tigecycline-TG, 1%) were determined in the treatment of Pythium insidiosum keratitis in rabbits. Infection of right eye of 38 rabbits was induced by standard intracorneal injection of P. insidiosum zoospores (left eye, intracorneal saline). Corneal infection developed in all right eyes. One hourly eye drops of one of the three antibiotics was instilled in both eyes (3 groups of 12 rabbits each) except in controls. Half of the rabbits in each group received intracorneal injection of the respective antibiotic after 4 days of starting eye drops. Clinical scoring of eyes was done over next 3 weeks. The reduction in scores post-treatment was significant for each drug (LZ: p < 0.025, AZ: p < 0.025, TG: p < 0.01). Scores with LZ (median change of 3) was significantly (p = 0.013) higher than TG (median change of 2) and comparable (p = 0.06) to AZ (median change of 3). Reduction in clinical scores in eyes receiving intracorneal antibiotics was not significantly different from the eyes that did not receive intracorneal antibiotics (p = 0.73). While no adverse effect of LZ was seen in the control corneas, 66-100% of rabbits showed reaction to AZ and TG. Histopathology showed severe inflammation in all infected corneas and intraocular extension in some of the rabbits with poor response. The success rate was noted to be 16.7%, 25% and 50% in AZ, TG and LZ respectively (p = 0.45). LZ demonstrated superior efficacy and safety and can be considered for trial in human disease.

'Targeting the feast of a sleeping beast': Nutrient and mineral dependencies of encysted Acanthamoeba castellanii.

Acanthamoeba spp. cause a corneal infection, Acanthamoeba keratitis (AK), and a cerebral infection, granulomatous amoebic encephalitis (GAE). Though aggressive chemotherapy has been able to kill the active trophozoite form of Acanthamoeba, the encysted form of this parasite has remained problematic to resist physiological concentrations of drugs. The emergence of encysted amoeba into active trophozoite form poses a challenge to eradicate this parasite. Acanthamoeba trophozoites have active metabolic machinery that furnishes energy in the form of ATPs by subjecting carbohydrates and lipids to undergo pathways including glycolysis and beta-oxidation of free fatty acids, respectively. However, very little is known about the metabolic preferences and dependencies of an encysted trophozoite on minerals or potential nutrients that it consumes to live in an encysted state. Here, we investigate the metabolic and nutrient preferences of the encysted trophozoite of Acanthamoeba castellanii and the possibility to target them by drugs that act on calcium ion dependencies of the encysted amoeba. The experimental assays, immunostaining coupled with bioinformatics tools show that the encysted Acanthamoeba uses diverse nutrient pathways to obtain energy in the quiescent encysted state. These findings highlight potential pathways that can be targeted in eradicating amoebae cysts successfully.

Canine trypanosomosis in Sri Lanka: An emerging problem reported from three distinct geographic locations.

Here we report three cases of canine trypanosomosis presented to the Veterinary Teaching Hospital in the Faculty of Veterinary Medicine and Animal Sciences at the University of Peradeniya, Sri Lanka during 2018. The cases were presented to the hospital when the dogs were already in critical condition. Confirmation of the cases was done by microscopic examination of Giemsa-stained thin blood smears. All three dogs had bilateral keratitis and anterior chamber cloudiness in eyes. Despite the intramuscular administration of diminazine aceturate, all of them subsequently died. Amplification and sequencing of a fragment of the internal transcribed spacer 1 (ITS1) of the nuclear ribosomal DNA confirmed the parasite as Trypanosoma. evansi. This is the first record of clinical cases of canine trypanosomosis in Sri Lanka. The three cases reported here came from widely separated geographical locations within the country: Balangoda, Mullaitivu and Kadawatha.

Infectious keratitis in southern Brazil: a comparison culture negative and culture positive patients / Ceratite infecciosa no sul do Brasil: comparação entre pacientes com cultura negativa e positiva

Abstract Purpose: To compare clinical-epidemiological profile and treatment outcome between culture negative and culture positive keratitis patients. Methods: Patients with suspected infectious keratitis seen at two ophthalmic hospitals in Curitiba, Brazil, between June 2014 and April 2016, were prospectively studied. Ophthalmological exam with corneal scraping and microbiological tests were performed. Data regarding follow up, surgical interventions and treatment outcome were collected after 12 weeks of the first visit trough medical chart review. From the results of the culture, two groups were formed: culture negative keratitis (CNK) and culture positive keratitis (CPK). Results: According to inclusion criteria 21 patients were classified as culture negative keratitis and 20 patients as culture positive keratitis. The number of patients on antibiotic drops at the first visit was greater in CNK group (90.5% versus 60%; p=0.032). Surgical procedures were necessary in 3 patients (15%) in CNK group and in 7 patients (36,8%) in CPK group (p=0.155). Treatment success was achieved by 85% (17/20) of the patients in CNK group and by 61% (11/18) of the patients in CPK group (p=0.144). There was no significant difference between groups regarding age, gender, place of residence, presence of comorbidities, risk factors for infectious keratitis, duration of symptoms and characteristics of corneal ulcer. Conclusions: Previous treatment with antibiotics correlates with negative culture results. There was no significant difference in treatment outcome between culture negative and culture positive keratitis patients.

Resumo Objetivo: Comparar os perfis clinico-epidemiológicos e os desfechos entre pacientes com ceratite com cultura positiva e pacientes com ceratite com cultura negativa. Métodos: Pacientes com ceratite infecciosa, atendidos em dois hospitais oftalmológicos em Curitiba, Brasil, entre junho de 2014 e abril de 2016, foram estudados prospectivamente. Exame oftalmológico, raspado de córnea e exames microbiológicos foram realizados no primeiro atendimento. Os dados quanto a seguimento e desfecho foram coletados após 12 semanas do primeiro atendimento através de revisão de prontuário. A partir dos resultados das culturas, dois grupos foram formados: ceratite com cultura negativa e ceratite com cultura positiva. Resultados: Vinte e um pacientes foram classificados como ceratite com cultura negativa e 20 como ceratite com cultura positiva. O número de pacientes em uso de colírio antibiótico no primeiro atendimento foi maior no grupo de cultura negativa (90,5% versus 60%; p=0,032). Sete pacientes (37%) no grupo cultura positiva precisaram de procedimentos cirúrgicos no manejo da ceratite, versus 3 pacientes (15%) do grupo cultura negativa (p=0,155). Oitenta e cinco por cento (17/20) dos pacientes do grupo cultura negativa alcançaram sucesso no tratamento, contra 61% (11/18) dos pacientes no grupo cultura positiva (p=0,144). Não houve diferença entre os grupos quanto a idade, gênero, local de procedência, presença de comorbidades, fatores de risco, duração dos sintomas e características da úlcera de córnea. Conclusão: Tratamento prévio com colírio de antibiótico correlaciona-se com resultados negativos de cultura. Não houve diferença no desfecho após tratamento entre os pacientes com cultura negativa e cultura positiva.

Bristle Worm-Induced Keratouveitis: A Case Report.

PURPOSE: To report keratouveitis arising from corneal penetration by a bristle worm seta (bristle). METHODS: We report on a 64-year-old woman presenting with uniocular pain, redness, reduced vision, discharge, and pustular rash of the hands after cleaning out her marine aquarium containing bristle worms and rubbing her eye. RESULTS: Slit-lamp biomicroscopy showed pseudomembranous conjunctivitis, corneal punctate erosions, stromal infiltrate, and edema, but no visible foreign body. The anterior chamber developed 3+ cells with hypopyon within 24 hours. Laboratory testing of corneal and hand samples was negative. In vivo confocal microscopy revealed a hair-like seta in the anterior corneal stroma, 25 to 105 µm wide, with surrounding inflammatory cells. Anterior chamber washout was performed. Topical prednisolone 0.5% was commenced, and corneal edema gradually resolved over 3 months. CONCLUSIONS: Bristle worms thrive as detritivores in marine aquarium sediment. Aquarium owners risk touching their setae during tank cleaning. Ophthalmia nodosa describes ocular reaction to caterpillar, vegetable, and spider hairs: features include posterior migration of hairs, granulomatous inflammation, and uncertain prognosis. Eye specialists should be alerted to the possibility of bristle worm-induced ophthalmia nodosa. Confocal microscopy may be useful in identifying microscopic setae.

Enterobious vermicularis induced infectious keratitis following penetrating keratoplasty.

A 28-year-old man with a prior history of penetrating keratoplasty in the left eye for total corneal melt presented with a 3-day history of a red, painful left eye. On examination, he was found to have graft infiltrates. Microscopic examination of wet mount preparation of corneal scrapings revealed the presence of embryonated eggs of E. vermicularis. Cellophane (scotch) tape preparation from perianal region also revealed embryonated eggs of E. vermicularis as well as live adult worms. Ocular and adnexal involvement by E. vermicularis is extremely rare. Here, we report the first case of infection of a corneal graft with E. vermicularis.

Antimicrobial Photodynamic Therapy with the photosensitizer TONS504 eradicates Acanthamoeba.

BACKGROUND: Microbial keratitis is a potential cause of corneal blindness. We investigated the amoebicidal efficacy of photodynamic antimicrobial therapy with a light-emitting diode as the light source and the cationic chlorin derivative TONS504 as the photosensitizer for the elimination of Acanthamoeba, a causative organism of corneal infection and blindness. Acanthamoeba keratitis remains a challenge to treat because of limited available treatments. METHODS: Acanthamoeba castellani 50370 was exposed to TONS504 at various concentrations (0, 1, or 10 mg/L for trophozoites; 0, 1, 10, or 20 mg/L for cysts), irradiated at various light energies (0, 10, or 30 J/cm2 for trophozoites; 0, 30, or 60 J/cm2 for cysts), and incubated at 26 °C for 3 h. Assessment of cell viability by trypan blue staining revealed that photodynamic antimicrobial therapy attenuated the survival of trophozoites and cysts dependent on TONS504 concentration and light energy. RESULTS: Photodynamic antimicrobial therapy with 10 mg/L TONS504 and 30 J/cm2 light energy suppressed trophozoite viability by 77%, and 20 mg/L TONS504 and 60 J/cm2 light energy attenuated cyst survival by 42%. Staining with fluorescein isothiocyanate-conjugated annexin V and ethidium homodimer III revealed photodynamic antimicrobial therapy induced apoptosis and necrosis in trophozoites dependent upon the intensity of treatment, whereas apoptosis was the predominant form of cell death in cysts. CONCLUSIONS: Photodynamic antimicrobial therapy with TONS504 warrants further investigation as a potential treatment modality for Acanthamoeba keratitis.

The most abundant cyst wall proteins of Acanthamoeba castellanii are lectins that bind cellulose and localize to distinct structures in developing and mature cyst walls.

BACKGROUND: Acanthamoeba castellanii, which causes keratitis and blindness in under-resourced countries, is an emerging pathogen worldwide, because of its association with contact lens use. The wall makes cysts resistant to sterilizing reagents in lens solutions and to antibiotics applied to the eye. METHODOLOGY/PRINCIPAL FINDINGS: Transmission electron microscopy and structured illumination microscopy (SIM) showed purified cyst walls of A. castellanii retained an outer ectocyst layer, an inner endocyst layer, and conical ostioles that connect them. Mass spectrometry showed candidate cyst wall proteins were dominated by three families of lectins (named here Jonah, Luke, and Leo), which bound well to cellulose and less well to chitin. An abundant Jonah lectin, which has one choice-of-anchor A (CAA) domain, was made early during encystation and localized to the ectocyst layer of cyst walls. An abundant Luke lectin, which has two carbohydrate-binding modules (CBM49), outlined small, flat ostioles in a single-layered primordial wall and localized to the endocyst layer and ostioles of mature walls. An abundant Leo lectin, which has two unique domains with eight Cys residues each (8-Cys), localized to the endocyst layer and ostioles. The Jonah lectin and glycopolymers, to which it binds, were accessible in the ectocyst layer. In contrast, Luke and Leo lectins and the glycopolymers, to which they bind, were mostly inaccessible in the endocyst layer and ostioles. CONCLUSIONS/SIGNIFICANCE: The most abundant A. castellanii cyst wall proteins are three sets of lectins, which have carbohydrate-binding modules that are conserved (CBM49s of Luke), newly characterized (CAA of Jonah), or unique to Acanthamoebae (8-Cys of Leo). Cyst wall formation is a tightly choreographed event, in which lectins and glycopolymers combine to form a mature wall with a protected endocyst layer. Because of its accessibility in the ectocyst layer, an abundant Jonah lectin is an excellent diagnostic target.

Biofilm testing of microbiota: An essential step during corneal scrap examination in Egyptian acanthamoebic keratitis cases.

PURPOSE: To detect co-infections in the culture-proven acanthamoebic keratitis (AK) cases, and to test the capability of biofilm formation in the isolated microbiota. The clinical findings, habit of wearing contact lens and in-vitro antibiotic resistance were analyzed further according to the biofilm formation capability. METHODS: After clinical examination, corneal scraps and swabs were taken from 240 clinically suspected AK cases, for Acanthamoeba and microbiological cultures. In cases of keratoplasty, trimmed corneal tissue was collected and sent for histopathological examination. Scanning electron microscopy was done for some samples. Biofilm formation capability was investigated using a tissue culture plate method. Antibiotic resistance pattern was determined using a modified-Kirby-Bauer disc diffusion method. RESULTS: In 102 AK culture proven cases, 11 had no co-infection, 74 had a single co-infection and 17 had double co-infections. Enterobactericae and Aspergillus were the commonest bacterial and fungal isolates, respectively. Regarding the biofilm formation, 64.7% of Enterobactericae, 50% of Pseudomonas aeuroginosa, 43.75% of Staph aureus, 76.92% of Streptococcus pneumoniae, 28.57% of Corynebacterium, 60% of α-haemolytic streptococci, 40% of Acinetobacter, 100% of Candida and 77.8% Aspergillus isolates were biofilm producers. Severe manifestations were more frequently reported in cases co-infected with biofilm producers than with non-biofilm producers. Generally, high percentages of the biofilm forming bacterial isolates were sensitive to antibiotics in-vitro. CONCLUSIONS: Routine investigations for co-infection and biofilm formation in addition to Acanthamoeba culture are strongly recommended in suspected AK cases. Co-infection with biofilm producers may precipitate extrinsic in-vivo drug resistance despite of the in-vitro sensitivity. Designing a biofilm-dissolving topical drug is highly recommended to enhance the response to the standard therapeutic regimen especially in the resistant AK cases.

Pythium Keratitis Leading to Fatal Cavernous Sinus Thrombophlebitis.

PURPOSE: To report a case of Pythium insidiosum keratitis leading to fatal cavernous sinus thrombophlebitis. METHODS: Case report. RESULTS: A 70-year-old man presented with excruciating pain, redness, and diminution of vision in his left eye for 2 weeks after washing his hair with tap water. A total corneal ulcer with surrounding infiltrates and associated corneal thinning was present. Corneal scraping revealed the presence of Gram-positive cocci. KOH wet mount and in vivo confocal microscopy revealed branching hyphae. Combined antibacterial and antifungal treatment was started, but 4 days later, the ulcer showed signs of worsening with perforation for which a large therapeutic penetrating keratoplasty was done. The host cornea showed branching septate hyphae on Sabarouds Dextrose Agar. Two weeks later, the patient developed left eye proptosis with associated extraocular movement restriction. Magnetic resonance imaging of the head and orbit revealed cavernous sinus thrombophlebitis. Lid sparing partial exenteration was performed. Polymerase chain reaction revealed P. insidiosum. The patient subsequently developed a cerebrovascular attack and died of its complications. CONCLUSIONS: Ocular pythiosis may lead to cavernous sinus thrombophlebitis and can even be life threatening. Timely diagnosis and early radical surgery are of value. A high index of suspicion must be kept for P. insidiosum in cases with suspected fungal etiology not responding to conventional treatment.

Keratitis After Post-Kala-Azar Dermal Leishmaniasis.

PURPOSE: To report 2 cases of keratitis occurring after post-kala-azar dermal leishmaniasis (PKDL) in immunocompetent patients. METHODS: Case report and literature review. RESULTS: Two male patients presented with a dense stromal infiltrate in the temporal and superior aspects of the cornea, respectively. Microbiological workup in both patients did not show any offending organism. Both patients were undergoing treatment for PKDL in the form of oral miltefosine. After careful deliberation, topical prednisolone was started under antibiotic cover for both patients with close follow-up. There was a dramatic improvement with resolution of the infiltrate and a good visual outcome. CONCLUSIONS: We present the first reported cases of keratitis after PKDL, which responded to topical corticosteroid therapy. The treating clinician must be made aware of such a presentation of leishmaniasis especially in endemic areas.

Significant correlation between meibomian gland dysfunction and keratitis in young patients with Demodex brevis infestation.

AIMS: To report the clinical characteristics and correlation between meibomian gland dysfunction (MGD) and keratitis in young patients with ocular demodicosis. METHODS: Observational case series of 60 patients younger than 35 years with ocular demodicosis, of which the diagnosis was based on microscopic counting of Demodex folliculorum and D. brevis of epilated lashes. Severity of keratitis and MGD was graded by photography and meibography, respectively, in a masked fashion. RESULTS: MGD was detected in 54/60 (90%) patients with the loss of meibomian gland in the upper lid more than the lower lid (p<0.001). Blepharoconjunctivitis and a variety of corneal pathologies were noted in 47/60 (78.3%) and 39/60 (65%) patients, respectively. For a total of 120 eyes, normal cornea was noted in 53 (44.2%) eyes, superficial punctate keratitis or limbitis was noted in 17 (14.2%), while corneal stromal infiltration was found in 50 (41.7%) eyes. Both univariate and multivariate analyses showed that the severity of meibomian gland loss was significantly correlated with higher D. brevis count and more severe keratitis (all p<0.05). Rapid resolution of keratitis and blepharoconjunctivitis was accompanied by significant reduction of the Demodex count in 48 patients receiving lid scrub directed to kill mites. CONCLUSIONS: There is a significant correlation between MGD and keratitis in young patients with ocular demodicosis especially inflicted by D. brevis.

Keratitis due to microfilariae in dogs: a newly recognized disease.

Parasitic agents have been associated with keratitis, but a diagnosis of parasitic keratitis has not been commonly made in domestic animals. The purpose of this study was to describe the clinical and histopathological findings in seven dogs with chronic keratitis caused by microfilariae diagnosed in Brazil. All dogs presented with superficial corneal opacities of varying degrees affecting the perilimbal and central regions of the cornea, with other opaque areas appearing as crystalline deposits and corneal vascularization. The lesions were bilateral and were associated with mild-to-moderate conjunctival hyperemia. There was no history of blepharospasm or pruritus, and no subjects presented with epithelial erosions. Corneal biopsy revealed free microfilariae in the corneal stroma, with varying degrees of inflammation and collagen fiber destruction. The microfilariae were also found in skin lesions by skin snip technique. No adult worms were found in these dogs, and no dogs were on heartworm preventative before diagnosis. Monthly doses of oral ivermectin improved ocular and dermal lesions. One dog showed complete remission with the treatment. The species of the microfilariae was not identified.

Status of the effectiveness of contact lens disinfectants in Malaysia against keratitis-causing pathogens.

The aim of this study was (i) to assess the antimicrobial effects of contact lens disinfecting solutions marketed in Malaysia against common bacterial eye pathogens and as well as eye parasite, Acanthamoeba castellanii, and (ii) to determine whether targeting cyst wall would improve the efficacy of contact lens disinfectants. Using ISO 14729 Stand-Alone Test for disinfecting solutions, bactericidal and amoebicidal assays of six different contact lens solutions including Oxysept®, AO SEPT PLUS, OPTI-FREE® pure moist®, Renu® fresh™, FreshKon® CLEAR and COMPLETE RevitaLens™ were performed using Manufacturers Minimum recommended disinfection time (MRDT). The efficacy of contact lens solutions was determined against keratitis-causing microbes, namely: Pseudomonas aeruginosa, Methicillin-resistant Staphylococcus aureus, Streptococcus pyogenes, Streptococcus pneumoniae, and Acanthamoeba castellanii. In addition, using chlorhexidine as an antiamoebic compound and cellulase enzyme to disrupt cyst wall structure, we determined whether combination of both agents can enhance efficacy of marketed contact lens disinfectants against A. castellanii trophozoites and cysts, in vitro. The results revealed that all contact lens disinfectants tested showed potent bactericidal effects exhibiting 100% kill against all bacterial species tested. In contrast, none of the contact lens disinfectants had potent effects against Acanthamoeba cysts viability. When tested against trophozoites, two disinfectants, Oxysept Multipurpose and AO-sept Multipurpose showed partial amoebicidal effects. Using chlorhexidine as an antiamoebic compound and cellulase enzyme to disrupt cyst wall structure, the findings revealed that combination of both agents in contact lens disinfectants abolished viability of A. castellanii cysts and trophozoites. Given the inefficacy of contact lens disinfectants tested in this study, these findings present a significant concern to public health. These findings revealed that targeting cyst wall by using cyst wall degrading molecules in contact lens disinfecting solutions will enhance their efficacy against this devastating eye infection.