Contact Lens Disinfection Research Articles

In vitro effects of multi-purpose contact lens disinfecting solutions towards survivability of Acanthamoeba genotype T4 in Malaysia.

The incidence of Acanthamoeba keratitis has been increasing since the previous decades, especially among contact lens users. This infection is majorly caused by the use of ineffective contact lens disinfecting solution. Thus, this study was conducted to evaluate the in vitro effects of multi-purpose disinfecting solutions (MPDS) against Acanthamoeba trophozoites and cysts. Acanthamoeba genotype T4 isolated from contact lens paraphernalia and an environmental strains were propagated for trophozoite or cyst-containing culture and adjusted in final concentration of 1 × 105 cells/ml. Amoebicidal and cysticidal assays were conducted by incubating trophozoites and cysts with OPTI-FREE® Express®, ReNu® Fresh™, Complete® Multi-Purpose Solution and AVIZOR Unica® Sensitive according to the manufacturer’s minimum recommended disinfectant time (MMRDT) for up to 12 h at 30 ⁰C. Trypan blue hemocytometer-based microscopic counts determined amoebicidal and cysticidal effects. The viability of Acanthamoeba trophozoites and cysts was confirmed by re-inoculated them in the 1.5% non-nutrient agar plates. It was found that none of the MPDS showed amoebicidal and cysticidal effects during the MMRDT. However, OPTI-FREE® Express® demonstrated a significant differences in average cell reduction for both stages within MMRDT. When subjected to 12 h exposure, both OPTI-FREE® Express® and ReNu® Fresh™ led to significant reduction in the number of trophozoite and cyst cells. Notably, Complete® Multi-Purpose Solution and AVIZOR Unica® Sensitive did appreciably improve the solution effectiveness towards trophozoite cells when incubated for 12 h. All MPDS were largely ineffective, with 100% survival of all isolates at MMRDT, while OPTI-FREE® Express® showed limited amoebicidal activity against the contact lens paraphernalia isolate, however, it was more against the environmental strains after 12 h incubation time. The commercially available MPDS employed in this research offered minimal effectiveness against the protozoa despite the contact time. Improvement or development of new solution should consider the adjustment of the appropriate disinfectant concentration, adequate exposure time or the incorporation of novel chemical elements, which are effective against Acanthamoeba for accelerated disinfecting and more reduction of potential exposure of contact lens users to Acanthamoeba keratitis.

Hydrogen Peroxide Disinfecting Solution for Gas Permeable Contact Lenses: A Review of the Antimicrobial Efficacy, Compatibility, and Safety Performance of a One-Step Lens Care System.

CLEAR CARE PLUS (CCP), also known as AOSEPT PLUS with HydraGlyde, is approved for use with gas permeable (GP) lenses, and the indication is supported by the scientific evidence that is reviewed in this article. Antimicrobial efficacy testing of CCP both as a stand-alone disinfectant and as part of a regimen shows that CCP exceeds the ISO 14729 criteria against bacteria, yeast, and mold. In real-world conditions, it is effective against clinically relevant bacterial strains isolated from adverse events and against the two forms, trophozoites and cysts, of resilient Acanthamoeba species. Compatibility tests of CCP with two types of GP lenses indicate that the physical and/or optical parameters of lenses are unaffected through 30 cycles of simulated use with CCP, and a clinical trial shows substantial equivalence of clinical performance with a commonly used GP multipurpose solution. These results indicate that CCP is well suited for cleaning and disinfection of GP contact lenses.

Tannic acid-modified silver nanoparticles enhance the anti-Acanthamoeba activity of three multipurpose contact lens solutions without increasing their cytotoxicity

BackgroundFree-living amoebae of the genus Acanthamoeba are cosmopolitan, widely distributed protozoans that cause a severe, vision-threatening corneal infection known as Acanthamoeba keratitis (AK). The majority of the increasing number of AK cases are associated with contact lens use. Appropriate eye hygiene and effective contact lens disinfection are crucial in the prevention of AK because of the lack of effective therapies against it. Currently available multipurpose contact lens disinfection systems are not fully effective against Acanthamoeba trophozoites and cysts. There is an urgent need to increase the disinfecting activity of these systems to prevent AK infections. Synthesized nanoparticles (NPs) have been recently studied and proposed as a new generation of anti-microbial agents. It is also known that some plant metabolites, including tannins, have anti-parasitic activity. The aim of this study was to evaluate the anti-amoebic activity and cytotoxicity of tannic acid-modified silver NPs (AgTANPs) conjugated with selected multipurpose contact lens solutions.MethodsThe anti-amoebic activities of pure contact lens care solutions, and NPs conjugated with contact lens care solutions, were examined in vitro by a colorimetric assay based on the oxido-reduction of alamarBlue. The cytotoxicity assays were performed using a fibroblast HS-5 (ATCC CRL-11882) cell line. The results were statistically analysed by ANOVA and Student-Newman-Keuls test using P < 0.05 as the level of statistical significance.ResultsWe show that the NPs enhance the anti-Acanthamoeba activities of the tested contact lens solutions without increasing their cytotoxicity profiles. The activities are enhanced within the minimal disinfection time recommended by the manufacturers.ConclusionsThe conjugation of the selected contact lens solutions with AgTANPs might be a novel and promising approach for the prevention of AK infections among contact lens users.Graphical

Enhancement of Contact Lens Disinfection by Combining Disinfectant with Visible Light Irradiation.

Multiple use contact lenses have to be disinfected overnight to reduce the risk of infections. However, several studies demonstrated that not only microorganisms are affected by the disinfectants, but also ocular epithelial cells, which come into contact via residuals at reinsertion of the lens. Visible light has been demonstrated to achieve an inactivation effect on several bacterial and fungal species. Combinations with other disinfection methods often showed better results compared to separately applied methods. We therefore investigated contact lens disinfection solutions combined with 405 nm irradiation, with the intention to reduce the disinfectant concentration of ReNu Multiplus, OptiFree Express or AOSept while maintaining adequate disinfection results due to combination benefits. Pseudomonads, staphylococci and E. coli were studied with disk diffusion assay, colony forming unit (cfu) determination and growth delay. A log reduction of 4.49 was achieved for P. fluorescens in 2 h for 40% ReNu Multiplus combined with an irradiation intensity of 20 mW/cm2 at 405 nm. For AOSept the combination effect was so strong that 5% of AOSept in combination with light exhibited the same result as 100% AOSept alone. Combination of disinfectants with visible violet light is therefore considered a promising approach, as a reduction of potentially toxic ingredients can be achieved.

In Vitro Amoebicidal Activity of Imidazolium Salts Against Trophozoites.

Several strains of the free-living genus Acanthamoeba can cause granulomatous amoebic encephalitis (GAE), a rare chronic and slowly progressive infection of the central nervous system (CNS), and Acanthamoeba keratitis (AK), a sight-threatening eye infectious disease. AK incidence has increased with the popularization of the contact lens wear and its treatment is currently limited and frequently unsuccessful. As imidazolium salts (IS), cationic imidazole derivatives, have promising antimicrobial potential. The present study evaluated the amoebicidal activity of four IS; 1-n-hexadecyl-3-methylimidazolium methanesulfonate (C16MImMeS), chloride (C16MImCl) and bis (triluoromethylsulfonyl) imide (C16MImNTf2 ), and 1-methyl-3-n-octadecylimidazolium chloride (C18MImCl), against the Acanthamoeba castellanii (ATCC30010) environmental strain and a clinical isolate (genotype T4). Three IS showed being lethal to 100% of the Acanthamoeba trophozoites at the minimum inhibitory concentrations of 125 and 62.5 μg/mL (C16MImMeS), 31.25 and 62.5 μg/mL (C16MImCl), and 125 and 125 μg/mL (C18MImCl) for ATCC30010 and isolate T4, respectively. C16MImNTf2 did not demonstrate amoebicidal activity. All active IS caused the hemolysis of erythrocytes. The cytotoxic effect of the IS was tested in RAW macrophages and human brain microvascular endothelial cells, which demonstrated cytotoxicity in all concentrations tested against both cell lines. As a consequence, these IS with amoebicidal activity presented low selectivity index values (SI) (SI < 1.0), demonstrating lack of parasite selectivity. Thus, C16MImMeS, C16MImCl, and C18MImCl seem to hold greater promise as components for contact lens cleaning and disinfection solutions, instead of direct human application.

Combination of tert-butyl hydroperoxide with vorinostat induces cell death of Acanthamoeba through cell cycle arrest

Safety precautions prior to contact lens usage is essential for preventing Acanthamoeba keratitis. Contact lens disinfecting solutions containing 3% hydrogen peroxide (H2O2) are known to exert amoebicidal effect against Acanthamoeba. Yet, these solutions need to be neutralized to prevent ocular irritation, which consequently may result in incomplete disinfection. In this study, amoebicidal effect of tert-butyl hydroperoxide (tBHP) was investigated and its efficacy was compared to those of hydrogen peroxide (H2O2). H2O2 and tBHP showed dose dependent amoebicidal effect, however high concentration of these compounds demonstrated cytotoxicity in human corneal epithelial (HCE) cells. To reduce their cytotoxicity, the concentrations of both compounds were diluted to 50 μM and subsequently combined with 10 μM vorinostat to enhance amoebicidal effect. Addition of vorinostat induced high amoebicidal effect against Acanthamoeba trophozoites, even at low concentrations of H2O2 or tBHP. Cellular damage induced by combined treatment of H2O2 or tBHP with vorinostat in Acanthamoeba were determined by assessing cell cycle arrest and apoptosis via FACS analysis. While 50 μM H2O2 combined with 10 μM vorinostat showed 36.26% cytotoxicity on HCE cells during 24 h exposure, 50 μM tBHP with 10 μM vorinostat did not show cytotoxicity on HCE cells. These findings suggest that the application of tBHP and vorinostat for Acanthamoeba keratitis treatment and contact lens disinfection system is highly plausible.

Pseudomonas aeruginosa and microbial keratitis.

Pseudomonas aeruginosa, a versatile Gram-negative pathogen that can cause a wide range of infections, is the most common causative agent in cases of bacterial keratitis associated with contact-lens use. Corneal infections with P. aeruginosa often have poor clinical outcomes and can result in long and costly treatments. During the infection process, the pathogen exploits its large genome, encoding complex regulatory networks and a wide range of virulence factors, including motility and the secretion of various proteases and toxins. Although antibiotic resistance levels in the UK are low, higher levels have been seen in some other countries. In the face of increasing antibiotic resistance, alternative therapeutic approaches such as antivirulence strategies and phage therapy are being developed. There is increasing evidence to suggest that keratitis infections are associated with a phylogenetic subgroup of P. aeruginosa isolates carrying the gene encoding the potent cytotoxin exotoxin U, one of two mutually exclusive exotoxins secreted via the type III secretion system. The mechanisms behind this association are unclear, but understanding the genetic differences that predispose P. aeruginosa to cause corneal infections may allow for the development of targeted and more effective future treatments to reduce the morbidity of P. aeruginosa keratitis. In order to minimize the risk of severe P. aeruginosa eye infections, a wide range of contact-lens disinfection solutions are available. Constant exposure to biocides at a range of concentrations, from sub-inhibitory to inhibitory, could contribute to the development of resistance to both antibiotics and disinfectants.

Realisation and assessment of a low-cost LED device for contact lens disinfection by visible violet light.

This study presents a device for efficient, low-cost and eye-friendly overnight disinfection of contact lenses by visible violet light as an alternative to disinfection with biocide-containing solutions. Bacterial solutions with one Pseudomonas and one Staphylococcus strain each were irradiated for up to 8 h in commercial transparent contact lens cases by the presented light-emitting diode (LED) device. Samples were taken at different intervals and distributed on agar plates. The surviving bacteria were determined by counting of colony-forming units and compared to the specific requirements of the stand-alone test for contact lens disinfection of the hygiene standard ISO 14729. The concentration of both microorganisms was reduced by three orders of magnitude after less than 4 h of irradiation. The LED current and intensity have not yet been at maximum and could be further increased if necessary for other microorganisms. The presented device fulfils the requirement of the stand-alone test of the contact lens hygienic standard ISO 14729 for the tested Pseudomonas and Staphylococcus strains. According to literature data, the inactivation of Serratia marcescens, Candida albicans and Fusarium solani seems also possible, but may require increased LED current and intensity.

Biocidal Efficacy of a Hydrogen Peroxide Lens Care Solution Incorporating a Novel Wetting Agent.

To compare the antimicrobial effects of CLEAR CARE, a 3% hydrogen peroxide (H2O2) solution formulated for simultaneous cleaning, daily protein removal, disinfection, and storage of soft (hydrophilic) hydrogel, silicone hydrogel, and gas-permeable contact lenses, and CLEAR CARE PLUS, consisting of the 3% H2O2 solution plus a novel wetting agent, polyoxyethylene-polyoxybutylene (EOBO-21). Three lots each of the 2 solutions were incubated with 5 compendial microorganisms required by the Food and Drug Administration (FDA) 510(k) and International Organization for Standardization (ISO) 14729 stand-alone procedures, 4 clinical isolates of Gram-positive and Gram-negative bacteria, and trophozoites and cysts of 2 Acanthamoeba strains that are associated with microbial keratitis. Microbial loads were evaluated after disinfection and neutralization. Both solutions exceeded the FDA/ISO stand-alone primary criteria against Gram-positive and Gram-negative compendial bacteria, yeast, and mold after only 1.5-hr disinfection/neutralization. At the recommended minimum disinfection time, bacteria were reduced by 4.4 to 5.1 logs, yeast by 4.4 to 4.9 logs, and mold by 2.9 to 3.5 logs with and without organic soil. In addition, both solutions eliminated or effectively reduced populations of clinically relevant ocular bacterial isolates (4.5-5.0 logs), Acanthamoeba trophozoites (3.4-4.2 logs), and cysts (1.5-2.1 logs). Both solutions eliminated or reduced populations of FDA/ISO compendial bacteria and fungi as well as clinically relevant microorganisms and Acanthamoeba trophozoites and cysts. The addition of EOBO-21 to the 3% H2O2 lens care solution had no impact on antimicrobial activity.

Evaluation of in vitro effect of selected contact lens solutions conjugated with nanoparticles in terms of preventive approach to public health risk generated by Acanthamoeba strains.

Various Acanthamoeba species are free-living organisms widely distributed in the human environment. Amphizoic amoebae as facultative parasites may cause vision-threatening eye disease - Acanthamoeba keratitis, mostly among contact lens wearers. As the number of cases is increasing, and applied therapy often unsuccessful, proper hygienic measures and effective contact lenses disinfection are crucial for the prevention of this disease. Available contact lens solutions are not fully effective against amphizoic amoebae; there is a need to enhance their disinfecting activity to prevent amoebic infections. The use of developing nanotechnology methods already applied with success in the prevention, diagnostic and therapy of other infectious diseases might be helpful regarding amoebic keratitis. This study assesses the in vitro effect of selected contact lens solutions conjugated with nanoparticles against Acanthamoeba trophozoites. Three selected contact lens solutions conjugated with silver and gold nanoparticles in concentration of 0.25-2.5 ppm were used in vitro against the axenically cultured ATCC 30010 type Acanthamoeba castellanii strain. The anti-amoebic efficacy was examined based on the oxido-reduction of AlamarBlue. The cytotoxicity tests based on the measurement of lactate dehydrogenase (LDH) activity were performed using a fibroblast HS-5 cell line. Enhancement of the anti-amoebic activity of contact lens solutions conjugated with selected nanoparticles expressed in the dose dependent amoebic growth inhibition with a low cytotoxicity profile was observed. Results of the study showed that conjugation of selected contact lens solutions with silver nanoparticles might be a promising approach to prevent Acanthamoeba keratitis among contact lens users.

Study of the Effectiveness of Multipurpose Solutions on the Bacterial Disinfection of Silicone Hydrogel Contact Lenses In Vitro

Objectives:To assess the antimicrobial effectiveness of multipurpose solutions in regard to the disinfection of silicone hydrogel contact lenses (CL) using a study of clinical bacterial isolates from ocular material.Methods:Three multipurpose solutions (solution A: polyhexamethylene biguanide 0.00025 g/100 mL; solution B: polyquaternary-1 0.001% and myristamidopropyl dimethylamine 0.0006%; and solution C: polyaminopropyl biguanide 0.00013% and polyquaternary 0.0001%) were used as a 3-phase disinfection on silicone hydrogel CL contaminated with bacteria from clinical isolates that were divided into five groups (group 1: Pseudomonas aeruginosa; group 2: Staphylococcus aureus; group 3: Staphylococcus epidermidis; group 4: Streptococcus spp; and group 5: enterobacteria).Results:No differences were observed between the 24- and 48-hr measurements in any of the samples, and the positivity of microorganisms in T0 was 100% for all solutions; it was 0% in T3. Therefore, only steps T1 (rubbing followed by rinsing) and T2 (rubbing followed by rinsing and immersion of CL into solution) were considered for analysis at the 24-hr measurement time. Throughout the phases, a decrease in the number of bacteria was observed, culminating in the elimination (no recovery) of all microorganisms in the three solutions.Conclusions:At the end of the proposed process, the tested solutions were effective.

Comparative in vitro effectiveness of a novel contact lens multipurpose solution on Acanthamoeba castellanii

BackgroundA multipurpose contact lens cleaning solution (MPS) containing novel active ingredients under development was compared to two commercially available MPS solutions for effectiveness against Acanthamoeba isolates.MethodsThe Acanthamoeba isolate A. castellanii was propagated for trophozoite or cyst-containing cultures for the purpose of assessment of effectiveness of each MPS. An alamar blue-based cellular respiration assay was used to assess effectiveness against trophozoites; Trypan blue hemocytometer-based microscopic counts measured cysticidal effects. To assess the general antimicrobial potency of each solution as controls for the anti-amoebic assays, comparative bactericidal effectiveness using Serratia marcenses was also performed.ResultsMinimal effectiveness against either Acanthamoeba form was observed from either commercial MPS. In contrast, the novel MPS achieved complete kill within 1 h contact time for both Acanthamoeba trophozoite and cysts. Each commercial MPS required 6 h contact time to achieve a two to three log reduction in S. marcenses. In contrast, the experimental MPS achieved disinfection in 60 min contact time, and complete kill (< 1 CFU) at 90 min.ConclusionsResults suggest that the inclusion of a novel ingredient combination within the MPS under development clearly is required and is ideal for rapid and effective killing of Acanthamoeba species in the context of contact lens disinfection systems. The representative commercially available MPS used in this testing provided minimal effectiveness against the protozoa regardless of contact time. In addition, comparative results with the bacterial agent in the control study show distinct differences in the speed to disinfection with the novel MPS. Future MPS development should consider inclusion of novel chemical entities that are effective against Acanthamoeba species to speed disinfection and further reduce the exposure potential of users of contact lenses and cleaning systems.

Study of Disinfectant Resistance Genes in Ocular Isolates of Pseudomonas aeruginosa.

Background: The prevalence of disinfectant resistance in Pseudomonas aeruginosa is on the rise. P. aeruginosa is the most common bacteria isolated from cases of microbial keratitis. Many multi-purpose contact lens disinfectant solutions are available to decontaminate contact lenses before use and to help reduce the incidence of infections. However, with increasing disinfectant resistance, the effect of multi-purpose disinfectant solutions may diminish. The goal of this study was to examine genes associated with disinfectant resistance in ocular isolates of P. aeruginosa and understand the strain’s susceptibility to different multipurpose disinfectant solutions. Methods: Seven potential disinfectant resistance genes were used in BLASTn searches against the whole genomes of 13 eye isolates of P. aeruginosa. A microdilution broth method was used to examine susceptibility to four different multipurpose disinfectant solutions. Results: All strains possessed the sugE2, sugE3 and emrE (qacE) genes. The sugE1 and qacEdelta1 genes were present in 6/13 isolates. No strains contained the qacF or qacG genes. All tested disinfectant solutions had the ability to kill all test strains at 100% concentration, with some strains being susceptible at 1:8 dilutions of the disinfecting solutions. However, the presence of disinfectant resistance genes was not associated with susceptibility to multi-purpose disinfectants. Conclusion: All four tested contact lens disinfectant preparations are effective against P. aeruginosa isolates regardless of the presence of disinfectant resistance genes.

Effect of 2, 6-Dichlorobenzonitrile on Amoebicidal Activity of Multipurpose Contact Lens Disinfecting Solutions.

Multipurpose contact lens disinfecting solutions (MPDS) are widely used to cleanse and disinfect microorganisms. However, disinfection efficacy of these MPDS against Acanthamoeba cyst remain insufficient. 2, 6-dichlorobenzonitrile (DCB), a cellulose synthesis inhibitor, is capable of increasing the amoebical effect against Acanthamoeba by inhibiting its encystation. In this study, we investigated the possibility of DCB as a disinfecting agent to improve the amoebicidal activity of MPDS against Acanthamoeba cyst. Eight commercial MPDS (from a to h) were assessed, all of which displayed insufficient amoebicidal activity against the mature cysts. Solution e, f, and h showed strong amoebicidal effect on the immature cysts. Amoebicidal efficacy against mature cysts remained inadequate even when the 8 MPDS were combined with 100 μM DCB. However, 4 kinds of MPDS (solution d, e, f, and h) including 100 μM DCB demonstrated strong amoebicidal activity against the immature cysts. The amoebicidal activity of solution d was increased by addition of DCB. Cytotoxicity was absent in human corneal epithelial cells treated with either DCB or mixture of DCB with MPDS. These results suggested that DCB can enhance the amoebicical activity of MPDS against Acanthamoeba immature cyst in vitro.

Acanthamoeba keratitis: confirmation of the UK outbreak and a prospective case-control study identifying contributing risk factors

Background/aimsAcanthamoeba keratitis (AK) is a chronic debilitating corneal infection principally affecting contact lens (CL) users. Studies were designed to test claims that the UK incidence may have increased in 2012–2014...

Microbiocidal characterization of a novel povidone-iodine based rigid contact lens disinfecting solution.

This study investigated the efficacy of a novel povidone-iodine based disinfection solution for rigid gas permeable (RGP) lenses compared to three other currently available multipurpose and hydrogen peroxide solutions. Bactericidal and fungicidal activities were assessed using ISO 14729 reference methods, whilst amoebicidal properties were evaluated using an automated viability counter. All solutions were also assessed for long term storage stability over a 3-month period. The FDA guidelines were adequately achieved by all solutions tested and 3-month storage did not affect their activity against bacterial and fungal agents. Activity against Acanthamoeba castellani trophozoites reached 1-log reduction for the povidone-iodine solution and approached this level for the remaining solutions. Activity against cysts was somewhat reduced and ranged from 78 to 86% reduction in viability. A povidone-iodine based solution, which may offer advantages as resistance to this agent has not been demonstrated and it lacks toxicity, provided equivalent antimicrobial activity to other RGP solutions.

Butan-1-ol as an extractant solvent in dispersive liquid-liquid microextraction in the spectrophotometric determination of aluminium

Determining aluminium ions at μg L−1 scale currently requires either costly analytical techniques such as inductively coupled plasma, and/or graphite furnace atomic absorption spectrometry. Dispersive liquid-liquid microextraction (DLLME) is designed to promote separation and preconcentration, thus making it possible to determine the analyte of interest without significant matrix influence. This study was aimed at the development of a spectrophotometric method to determine Al3+ after microextraction of its complex with quercetin. Butan-1-ol was used as a novel extractant solvent in the DLLME process. The parameters influencing complexation and microextraction, such as the amount of quercetin and volume of extractant were evaluated by univariate analysis. In optimised conditions were estimated for the proposed method: linear range from 7.5 to 165.0 μg L−1, LOD of 2.0 μg L−1, and LOQ of 7.0 μg L−1. The accuracy was checked by applying the proposed method to water (NIST SRM-1643e) and rice flour (NIST SRM-1568c) certified reference materials and spike-and-recovery trials with distinct samples (mineral water, green tea, thermal spring water, contact lens disinfecting solution, saline concentrate for hemodialysis and urine).

Comparative antimicrobial efficacy of a novel povidone iodine rigid gas permeable (RGP) contact lens disinfection system

Comparative antimicrobial efficacy of a novel povidone iodine rigid gas permeable (RGP) contact lens disinfection system

Comparative antimicrobial efficacy of a novel povidone iodine based contact lens disinfection system

Comparative antimicrobial efficacy of a novel povidone iodine based contact lens disinfection system

A new strategy for enhancing acanthamoebicidal activity with synthesis of nanoflower of Laurocerausus officinalis Roemer (cherry laurel) fruit extracts

Pathogenic Acanthamoeba species often cause infection known as Acanthamoeba keratitis among people who use contact lenses. It is a type of infection that can result in corneal ulceration, visual loss or even blindness, if not treated. There are various therapeutic options available in the treatment of Acanthamoeba infections but they are usually tough treatments with limited efficacy. For instance, hydrogen peroxide (H2O2) is a commonly used contact lens disinfectant which is effective against Acanthamoeba but it is toxic to the cornea. For these reasons, new and more efficacious treatment options are required for Acanthamoeba infections. In this context, plants are considered natural resources for the discovery of new drugs. Laurocerasus officinalis Roem. (cherry laurel) (Rosaceae) grows in Black Sea region; and it is known as "Taflan", "Laz kirazı" or "Karayemis". Local people are using the seeds against diabetes, while the fruits are consuming as food, and used fordiuretic and passing kidney stones. It has also been reported that the seeds of the cherry laurel are used as an antiparasitic agent in this area. The aim of the study was to confirm the traditionally use of antiparasitic activity of this fruit and to increase the potential effect by means of organic-inorganic hybrid synthesis. Total phenol contents of methanol extracts prepared from endocarp, mesocarp and seeds of the fruit were calculated. The effects of methanol extracts and nano flower (NFs) plants synthesized from these extracts on the proliferation of Acanthamoeba castellanii were investigated. Thus, for the first time, novel organic-inorganic nanobio-antiparasitic agents called NFs were produced from cherry laurel and the increase in the amoebicidal activity of the NFs was elucidated. The characterization of NFs were determined with Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectrometer (FT-IR) and Energy-Dispersive X-ray (EDX) techniques. In addition, the catalytic activity of the fruit extracts and the NFs were measured against guaiacol in the presence of H2O2. The viability testing of A.castellanii cysts used for amoebicidal activity was performed using 4% trypan blue. Methanol extracts and nano-flowers were prepared at concentrations of 32, 16, 8, 4, 2 and 1 mg/ml in 0.9% saline and distributed 200 μl each in tubes and incubated in the room temperature with the addition of 200 μl of 98% viable A.castellani parasites. The results were evaluated using the SPSS V.22.0 program and it was determined that there was a significant increase in the amoebicidal activity of NFs compared with the other extracts according to variance analysis (p≤ 0.05). In the study, it was determined that samples killed parasites or reduced parasite proliferation at certain times. As a result, NFs synthesized from fruit extracts were demonstrated about three times more effective than the non hybrid extracts for amoebicidal activity. This situation can be explained as high proliferative effect of a new nano-bio-antiparasitic agent known as nanoflower against A.castellanii.