scholarly journals Enhanced Killing of Acanthamoeba Cysts with a Plant Peroxidase-Hydrogen Peroxide-Halide Antimicrobial System

2003 ◽  
Vol 69 (5) ◽  
pp. 2563-2567 ◽  
Author(s):  
Reanne Hughes ◽  
Peter W. Andrew ◽  
Simon Kilvington
Keyword(s):  
Hydrogen Peroxide ◽  
Horseradish Peroxidase ◽  
Contact Lens ◽  
Acanthamoeba Keratitis ◽  
Free Living ◽  
Soybean Peroxidase ◽  
Plant Peroxidase ◽  
Time Points ◽  
Free Living Amoeba ◽  
Care Systems

ABSTRACT The activity of H2O2 against the resistant cyst stage of the pathogenic free-living amoeba Acanthamoeba was enhanced by the addition of KI and either horseradish peroxidase or soybean peroxidase or, to a lesser degree, lactoperoxidase. This resulted in an increase in the cysticidal activity of 3% (wt/vol) H2O2, and there was >3-log killing in 2 h, compared with the 6 h required for comparable results with the peroxide solution alone (P < 0.05). With 2% H2O2, enhancement was observed at all time points (P < 0.05), and total killing of the cyst inoculum occurred at 4 h, compared with 6 h for the peroxide alone. The activity of sublethal 1% H2O2 was enhanced to give 3-log killing after 8 h of exposure (P < 0.05). No enhancement was obtained when KCl or catalase was used as a substitute in the reaction mixtures. The H2O2 was not neutralized in the enhanced system during the experiments. However, in the presence of a platinum disk used to neutralize H2O2 in contact lens care systems, the enhanced 2% H2O2 system gave 2.8-log killing after 6 h or total cyst killing by 8 h, and total neutralization of the H2O2 occurred by 4 h. In contrast, 2% H2O2 alone resulted in <0.8-log killing of cysts in the presence of the platinum disk due to rapid (<1 h) neutralization of the peroxide. Our observations could result in significant improvement in the efficacy of H2O2 contact lens disinfection systems against Acanthamoeba cysts and prevention of acanthamoeba keratitis.

scholarly journals Continuous Real-Time Motility Analysis of Acanthamoeba Reveals Sustained Movement in Absence of Nutrients

Pathogens ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 995
Author(s):  
Allison Campolo ◽  
Valerie Harris ◽  
Rhonda Walters ◽  
Elise Miller ◽  
Brian Patterson ◽  
...  
Keyword(s):  
Contact Lens ◽  
Contact Lenses ◽  
Axenic Culture ◽  
Acanthamoeba Keratitis ◽  
Ocular Infection ◽  
Glass Coverslip ◽  
E Coli ◽  
Total Distance ◽  
Care Systems ◽  
Different Strains

Acanthamoeba keratitis is a serious ocular infection which is challenging to treat and can lead to blindness. While this pathogen is ubiquitous and can contaminate contact lenses after contact with water, its habits remain elusive. Understanding this organism’s natural behavior will better inform us on how Acanthamoeba colonize contact lens care systems. Acanthamoeba trophozoites were allowed to adhere to either a glass coverslip or non-nutrient agar (NNA) within a flow cell with nutrients (Escherichia coli or an axenic culture medium (AC6)) or without nutrients (Ringer’s solution). Images were taken once every 24 s over 12 h and compiled, and videos were analyzed using ImageJ Trackmate software. Acanthamoeba maintained continuous movement for the entire 12 h period. ATCC 50370 had limited differences between conditions and surfaces throughout the experiment. Nutrient differences had a noticeable impact for ATCC 30461, where E. coli resulted in the highest total distance and speed during the early periods of the experiment but had the lowest total distance and speed by 12 h. The Ringer’s and AC6 conditions were the most similar between strains, while Acanthamoeba in the E. coli and NNA conditions demonstrated significant differences between strains (p < 0.05). These results indicate that quantifiable visual tracking of Acanthamoeba may be a novel and robust method for identifying the movement of Acanthamoeba in relation to contact lens care products. The present study indicates that Acanthamoeba can undertake sustained movement for at least 12 h with and without nutrients, on both rough and smooth surfaces, and that different strains have divergent behavior.

scholarly journals Isoniazid Conjugated Magnetic Nanoparticles Loaded with Amphotericin B as a Potent Antiamoebic Agent against Acanthamoeba castellanii

Antibiotics ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 276
Author(s):  
Kawish Iqbal ◽  
Sumayah Abdelnasir Osman Abdalla ◽  
Ayaz Anwar ◽  
Kanwal Muhammad Iqbal ◽  
Muhammad Raza Shah ◽  
...  
Keyword(s):  
Magnetic Nanoparticles ◽  
Amphotericin B ◽  
Central Nervous System Infection ◽  
Acanthamoeba Castellanii ◽  
Acanthamoeba Keratitis ◽  
Free Living ◽  
Granulomatous Amoebic Encephalitis ◽  
Nano Drug Delivery ◽  
Free Living Amoeba ◽  
Acanthamoeba Species

The pathogenic free-living amoeba, Acanthamoeba castellanii, is responsible for a rare but deadly central nervous system infection, granulomatous amoebic encephalitis and a blinding eye disease called Acanthamoeba keratitis. Currently, a combination of biguanides, amidine, azoles and antibiotics are used to manage these infections; however, the host cell cytotoxicity of these drugs remains a challenge. Furthermore, Acanthamoeba species are capable of transforming to the cyst form to resist chemotherapy. Herein, we have developed a nano drug delivery system based on iron oxide nanoparticles conjugated with isoniazid, which were further loaded with amphotericin B (ISO-NPs-AMP) to cause potent antiamoebic effects against Acanthamoeba castellanii. The IC50 of isoniazid conjugated with magnetic nanoparticles and loaded with amphotericin B was found to be 45 μg/mL against Acanthamoeba castellanii trophozoites and 50 μg/mL against cysts. The results obtained in this study have promising implications in drug discovery as these nanomaterials exhibited high trophicidal and cysticidal effects, as well as limited cytotoxicity against rat and human cells.

scholarly journals Encystment Induces Down-Regulation of an Acetyltransferase-Like Gene in Acanthamoeba castellanii

Pathogens ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 321 ◽  
Author(s):  
Steven Rolland ◽  
Luce Mengue ◽  
Cyril Noël ◽  
Stéphanie Crapart ◽  
Anne Mercier ◽  
...  
Keyword(s):  
Bioinformatics Analysis ◽  
Acanthamoeba Castellanii ◽  
Acanthamoeba Keratitis ◽  
Free Living ◽  
Adverse Conditions ◽  
Genetic Modifications ◽  
Granulomatous Amoebic Encephalitis ◽  
Free Living Amoeba ◽  
Causative Agents

Acanthamoeba castellanii is a ubiquitous free-living amoeba. Pathogenic strains are causative agents of Acanthamoeba keratitis and granulomatous amoebic encephalitis. In response to adverse conditions, A. castellanii differentiate into cysts, which are metabolically inactive and resistant cells. This process, also named encystment, involves biochemical and genetic modifications that remain largely unknown. This study characterizes the role of the ACA1_384820 Acanthamoeba gene during encystment. This gene encodes a putative N-acetyltransferase, belonging to the Gcn5-related N-acetyltransferase (GNAT) family. We showed that expression of the ACA1_384820 gene was down-regulated as early as two hours after induction of encystment in A. castellanii. Interestingly, overexpression of the ACA1_384820 gene affects formation of cysts. Unexpectedly, the search of homologs of ACA1_384820 in the Eukaryota gene datasets failed, except for some species in the Acanthamoeba genus. Bioinformatics analysis suggested a possible lateral acquisition of this gene from prokaryotic cells. This study enabled us to describe a new Acanthamoeba gene that is down-regulated during encystment.

scholarly journals Aminoglucoside/Imidazole in the Treatment of Acanthamoeba Keratitis

2018 ◽  
Vol 2 (2) ◽  
Keyword(s):  
Visual Acuity ◽  
Acanthamoeba Keratitis ◽  
Final Visual Acuity ◽  
Free Living ◽  
Corneal Infection ◽  
Free Living Amoeba

Acanthamoeba Keratitis (AK) is an infrequent corneal infection caused by free living amoeba, it is frequently misdiagnosed and medically/surgical treated with low or no response in advanced cases. In this paper we present five cases of AK with early diagnoses and good response to aminoglucoside/imidazole treatment and achieving acceptable final visual acuity in each case.

scholarly journals Comparison of Hydrogen Peroxide Contact Lens Disinfection Systems and Solutions against Acanthamoeba polyphaga

2001 ◽  
Vol 45 (7) ◽  
pp. 2038-2043 ◽  
Author(s):  
Reanne Hughes ◽  
Simon Kilvington
Keyword(s):  
Hydrogen Peroxide ◽  
Contact Lens ◽  
Lens Protein ◽  
Sodium Perborate ◽  
Acanthamoeba Polyphaga ◽  
Log Reduction ◽  
Disinfection Process ◽  
Free Living Amoeba ◽  
One Step ◽  
Corneal Damage

ABSTRACT Acanthamoeba is a free-living amoeba causing a potentially blinding infection of the cornea. Contact lens wearers are most at risk and account for some 95% of cases. Hydrogen peroxide is used for contact lens disinfection due to its broad antimicrobial activity. Lenses must be neutralized before use to avoid pronounced stinging and possible corneal damage. Neutralization is achieved by adding a catalyst during the disinfection process (one-step) or afterwards (two-step). Here, the activities of commercial peroxide systems and individual solutions against trophozoites and cysts ofAcanthamoeba polyphaga were compared. All disinfection systems were active against trophozoites, giving a ≥3-log (99.9%) kill within 1 h. Of the four one-step systems, only one showed some cysticidal activity, giving a 1.28 ± 0.41-log reduction. Both two-step systems were cysticidal, giving a ≥3-log kill at 4 h. All system peroxide solutions were cysticidal, giving a ≥3-log kill by 4 to 6 h. Variation in the cysticidal rate was observed with two solutions that gave a 1.8- to 2.1-log kill at 4 h compared with 3.0 to 4.0 for the rest (P < 0.05). No cysticidal activity was found with the peroxigen sodium perborate or the contact lens protein remover subtilisin A. Two-step systems are cysticidal providing contact times of at least 4 h are employed. Variation in cyst killing occurs between peroxide solutions, possibly due to formulation differences. One-step systems are less effective againstAcanthamoeba cysts due to rapid peroxide neutralization. The cysticidal activity of one-step systems could be improved if neutralization rates were retarded.

scholarly journals Evaluating Alternate Methods of Determining the Antimicrobial Efficacy of Contact Lens Care Products against Acanthamoeba Trophozoites

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 126 ◽  
Author(s):  
Allison Campolo ◽  
Paul Shannon ◽  
Monica Crary
Keyword(s):  
Propidium Iodide ◽  
Contact Lens ◽  
Acanthamoeba Keratitis ◽  
Antimicrobial Efficacy ◽  
Ocular Infection ◽  
Outer Membranes ◽  
Log Reduction ◽  
Free Living Amoeba ◽  
Corneal Damage ◽  
Rapid Tool

Acanthamoeba keratitis (AK) is a serious ocular infection caused by a ubiquitous free-living amoeba, Acanthamoeba. This infection often results in extensive corneal damage and blindness, and is notoriously difficult to cure. While Acanthamoeba is an abundant organism, AK is most associated with contact lens hygiene noncompliance and inadequate contact lens care (CLC) disinfection regimens. Thus, accurate and timely antimicrobial efficacy testing of CLC solutions is paramount. Published methods for antimicrobial efficacy testing of Acanthamoeba trophozoites requires 14 days for results. Presently, alternate and/or rapid methods for evaluating CLC products rarely demonstrate equivalent results compared to commonly-reported methods. Propidium iodide is a cellular stain that can only bind to cells with damaged outer membranes. We evaluated propidium iodide staining as an alternative method for determining the relative antimicrobial efficacy of 11 different CLC products against Acanthamoeba trophozoites. Following exposure to a CLC product, the fluorescence intensity of propidium iodide in an Acanthamoeba population demonstrated a strong correlation to the log reduction determined by established, growth-based Acanthamoeba testing used to evaluate the antimicrobial efficacy of CLC products. Thus, propidium iodide was found to be an effective rapid tool for determining cell death in Acanthamoeba trophozoites following exposure to CLC solutions.

Acanthamoeba spp. in river water samples from the Black Sea region, Turkey

2020 ◽  
Vol 18 (2) ◽  
pp. 186-199 ◽  
Author(s):  
İlknur Koyun ◽  
Zeynep Kolören ◽  
Ülkü Karaman ◽  
Amalia Tsiami ◽  
Panagiotis Karanis
Keyword(s):  
Black Sea ◽  
Water Samples ◽  
Tap Water ◽  
Acanthamoeba Keratitis ◽  
The Black Sea ◽  
Free Living ◽  
Black Sea Region ◽  
River Waters ◽  
Free Living Amoeba ◽  
Acanthamoeba Species

Abstract The present study aims to investigate the occurrence of free living amoeba (FLA) in water resources (rivers and tap water) in Samsun in the Black Sea. The presence of Acanthamoeba spp. was confirmed in 98 of 192 water samples collected from 32 sites of Samsun province (Samsun centre, Terme, Carsamba, Tekkekoy, Bafra) by PCR. Acanthamoeba spp. were found in 15/36 river samples from Samsun, in 58/90 from Terme, in 12/30 from Carsamba, in 7/18 from Tekkekoy and in 6/18 from Bafra. No Acanthamoeba species were detected in tap water samples. The highest rate in river waters contaminated with Acanthamoeba species was in Terme followed by Samsun centre (41.7%), Carsamba (40%), Tekkekoy (38.9%) and Bafra districts (33.3%), respectively. The result of the subsequent sequence analysis showed Haplotype I (A. triangularis) in 5%, Haplotype II (A. polyphaga) in 29.6%, Haplotype III (Acanthamoeba spp.) in 62% and Haplotype IV (A. lenticulata) in 3%. The most common genotype was Acanthamoeba T4 (Acanthamoeba spp., A. polyphaga, A. triangularis) and T5 genotype was also found in 3%. The T4 genotype is the most common genotype associated with Acanthamoeba keratitis (AK) worldwide; therefore, humans and animals living in the area are at risk after contact with such waters.

Ophthalmology hospital wards contamination to pathogenic free living Amoebae in Iran

2015 ◽  
Vol 60 (3) ◽  
Author(s):  
Zohreh Lasjerdi ◽  
Maryam Niyyati ◽  
Jacob Lorenzo-Morales ◽  
Ali Haghighi ◽  
Niloofar Taghipour
Keyword(s):  
Acanthamoeba Keratitis ◽  
Slit Lamp ◽  
Free Living ◽  
Eye Surgery ◽  
Disinfection Methods ◽  
Eye Trauma ◽  
Morphological Criteria ◽  
Free Living Amoeba ◽  
Hospital Wards ◽  
Good Hygiene

AbstractThe present study was conducted to determine the occurrence of potentially pathogenic free-living amoeba in ophthalmology wards in reference hospitals in Iran. Since an increasing number of Acanthamoeba Keratitis cases after eye surgery and eye trauma have been recently observed in this country, it could be possible that the disinfection procedures undertaken in the clinical setting may not have a good hygiene and disinfection procedures, hence the aim of this study. Therefore, 42 dust and biofilm samples were collected from different areas of ophthalmology wards and checked for the presence of FLA using morphological criteria, PCR based analysis and DNA sequencing. Of the 42 samples from dust and biofilm sources, 18(42.86%) isolates were found to contain FLA and 12(92.3%) isolates belonged to Acanthamoeba T4 genotype. Isolation of the pathogenic genotype T4 from medical instruments, including slit lamp in corneal wards, may be a threat for patients undergoing eye surgery in these wards. Other FLA isolated in this study included Acanthamoeba genotype T5, Vahlkampfia sp, Naegleria australiensis, Vermamoeba vermiformis and Echinamoeba exudans. To our knowledge, this is the first report of the presence of potentially pathogenic FLA in ophthalmology wards in Iran. Improved disinfection methods and monitoring of hospitals ward are thus necessary in this area in order to minimize the risk of infection in patients.

scholarly journals Susceptibility of various corneal fungal isolates and Pseudomonas aeruginosa to contact lens disinfecting solutions

2013 ◽  
Vol 7 (03) ◽  
pp. 261-268 ◽  
Author(s):  
Antonio Pinna ◽  
Donatella Usai ◽  
Stefania Zanetti ◽  
Philip A Thomas ◽  
Jayaraman Kaliamurthy
Keyword(s):  
Hydrogen Peroxide ◽  
Pseudomonas Aeruginosa ◽  
Contact Lens ◽  
Luria Bertani ◽  
Sabouraud Agar ◽  
Fungal Strains ◽  
Soft Contact Lens ◽  
Fungal Isolates ◽  
Colony Forming Units ◽  
Care Systems

Introduction: We aimed to investigate the susceptibility of a combined inoculum of Pseudomonas aeruginosa and different fungal strains to 6 soft contact lens disinfectants. Methodology: One corneal isolate of P. aeruginosa and 13 corneal fungal isolates (9 Aspergillus spp, 3 Fusarium spp, 1 Curvularia sp.) were used. The following solutions were tested: Arion Cronos, Complete RevitaLens, Dua Elite, Opti-Free Express, Regard, Oxysept Comfort, and Oxysept Comfort without catalase. The effect of the solutions was assessed on a combined inoculum of P. aeruginosa plus 1 fungal strain. Suspensions of P. aeruginosa and fungi were made in the solutions (1x106 colony-forming units/mL). After 1 hour (Arion Cronos only), 6, 8, and 24 hours, aliquots of suspension were removed and seeded on Luria-Bertani and Sabouraud agar plates. Results: After 6 hours’ exposure, all the solutions but Dua Elite and Oxysept Comfort eradicated P. aeruginosa. Conversely, apart from 3% hydrogen peroxide-based Oxysept Comfort without catalase, which eradicated all the fungi tested after 6 hours, all the other solutions were partly ineffective at killing some of the fungal isolates, even after 24 hours’ exposure. Conclusions: Most contact lens disinfectants may be ineffective if contact lens care systems become co-contaminated with P. aeruginosa and fungi. In our experiment, only exposure to 3% hydrogen peroxide without neutralizer for at least 6 hours was always able to kill a combined inoculum of P. aeruginosa and different fungal strains.

scholarly journals Effect of a Commercial Disinfectant CLORICAN® on Acanthamoeba spp. and Naegleria fowleri Viability

Parasitologia ◽  
2021 ◽  
Vol 1 (3) ◽  
pp. 119-129
Author(s):  
Ines Sifaoui ◽  
Aitor Rizo-Liendo ◽  
María Reyes-Batlle ◽  
Iñigo Arberas-Jiménez ◽  
Rubén L. Rodríguez-Expósito ◽  
...  
Keyword(s):  
Chlorine Dioxide ◽  
Acanthamoeba Keratitis ◽  
Naegleria Fowleri ◽  
Free Living ◽  
Short Period ◽  
Free Living Amoeba ◽  
Swimming Pool Water ◽  
Microbial Proliferation ◽  
Nægleria Fowleri ◽  
Primary Amebic Meningoencephalitis

Swimming pool water treatment by chemicals is an essential step to avoid microbial proliferation and infections namely caused by free living amoeba such as, for example, primary amebic meningoencephalitis and Acanthamoeba keratitis. In the present study, a commercial reactive, CLORICAN, based on chlorine dioxide, was evaluated against Acanthamoeba spp. and Naegleria fowleri. We observed that CLORICAN could eliminate in a short period of incubation time both amoebae. Furthermore, Naegleria fowleri’s trophozoites were more sensitive than those of Acanthamoeba spp. By means of inverted microscopy, the chlorine dioxide was found to greatly affect morphology shape by increasing the cell size shrinkage.

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