Abstract
This study evaluated in vitro effect of different concentrations of Aloe vera (A. vera) ethanol extract and honey against Acanthamoeba spp. cysts in comparison with chlorhexidine (the drug of choice for treatment of Acanthamoeba infection) at different incubation periods. Four different concentrations of the tested agents were used, 100, 200, 400, and 600 μg/ml for A. vera ethanol extract and 25, 50, 100, and 200 μg/ml for honey. Isolated Acanthamoeba spp. cysts from keratitis patients were incubated with different concentrations of the tested agents as well as chlorhexidine 0.02% (drug control) for different incubation periods (24, 48, 72 h). After each incubation period, the effect of A. vera extract and honey against Acanthamoeba cysts was assessed by counting the number of viable cysts, determining the inhibitory percentage and detecting the morphological alternations of treated cysts compared to non-treated and drug controls. Both A. vera ethanol extract and honey showed a concentration and time-dependent effect on the viability of Acanthamoeba cysts. In comparison with chlorhexidine (the drug control), A. vera ethanol extract possessed a potent cysticidal activity at all tested concentrations throughout different incubation periods, except for concentration 100 μg/ml which recorded the lower inhibitory effect. With increasing the dose of A. vera ethanol extract to 200, 400, 600 µg/ml, the recorded inhibitory percentages of Acanthamoeba cysts viability were 82.3%, 92.9% and 97.9% respectively, after 72 h compared to 76.3% of chlorhexidine. Similarly, honey at concentrations of 50–100 µg/ml gave higher inhibitory effect of 59% and 76.7%, respectively compared to chlorhexidine which showed an inhibitory percentage of 55.7% after 24 h. Meanwhile, the lowest tested concentration of honey (25 µg/ml) gave an inhibitory effect by 47.7–67% which was less than that of chlorhexidine throughout different incubation periods. With increasing the dose of honey to 200 µg/ml, the inhibitory effect was 98.9% after 72 h higher than that of chlorhexidine (76.9%). Using a scanning electron microscope, Acanthamoeba cysts treated by A. vera ethanol extract showed alternations in their shapes with flattening, collapsing, and laceration of their walls. Also, treated cysts by honey were highly distorted and difficult to identify because most of them were shrinkage and collapsed to a tiny size. On the other hand, chlorhexidine showed less structural and morphological changes of Acanthamoeba cysts. A. vera ethanol extract and honey had considerable cysticidal effects on Acanthamoeba cysts. They may give promising results for treatment of Acanthamoeba keratitis.







Similar content being viewed by others
Data availability
The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.
References
Abdulrahman O, Samaila AB, Panda SM, Aliyu A, Sahal MR (2019) Screening for antimicrobial activities of methanolic extracts of Aloe vera and Hyptissuaveolens against co-infections of Giardia lamblia and Salmonella among diarrhoeagenic children. South Asian J Res Microbiol 4(2):1–6
Anwar A, Ting ELS, Anwar A, ul Ain N, Faizi S, Shah MR, Khan NA, Siddiqui R (2020) Anti-amoebic activity of plant-based natural products and their conjugated silver nanoparticles against Acanthamoeba castellanii (ATCC 50492). AMB Express 10(1):24
Badria FA, Hetta MH, Sarhan RM, Ezz ElDin MH (2014) Lethal effects of Helianthemum lippii (L.) on Acanthamoeba castellanii cysts in vitro. Korean J Parasitol 52(3):243–249
Behera HS, Satpathy G (2017) Identification of Acanthamoeba spp. with different microscopes and analysis of the anatomical changes from trophozoite to cyst form with an electron microscope. EC Microbiol 8(4):203–210
Bulus T, Ahmed AB, Aboi TY, Danbaki DA (2016) Determination of IC50 and IC90 values of ethanolic extracts of some medicinal plants against Trypanosoma brucei brucei. Arch Clin Microbiol 7(3):1–5
Curto EM, Labelle A, Chandler HL (2014) Aloe vera: an in vitro study of effects on corneal wound closure and collagenase activity. Vet Ophthalmol 17(6):403–410
Dodangeh S, Niyyati M, Kamalinejad M, Lorenzo-Morales J, Haghighi A, Azargashb E (2017) The amoebicidal activity of Ziziphus vulgaris extract and its fractions on pathogenic Acanthamoeba trophozoites and cysts. Trop Biomed 34(1):127–136
El-Sayed NM, Hikal WM (2015) Several staining techniques to enhance the visibility of Acanthamoeba cysts. Parasitol Res 114(3):823–830
El-Sayed NM, Younis MS, Elhamshary AM, Abd-Elmaboud AI, Kishik SM (2014) Acanthamoeba DNA can be directly amplified from corneal scrapings. Parasitol Res 113(9):3267–3272
El-Sayed NM, Ismail KA, Ahmed SA, Hetta MH (2012) In vitro amoebicidal activity of ethanol extracts of Arachis hypogaea L., Curcuma longaL. and Pancratium Maritimum L. on Acanthamoeba castellanii cysts. Parasitol Res 110(5):1985–1992
Gupta VK, Malhotra S (2012) Pharmacological attribute of Aloe vera: Revalidation through experimental and clinical studies. Ayu 33(2):193–196
Hadaś E, Derda M, Cholewiński M (2017) Evaluation of the effectiveness of tea tree oil in treatment of Acanthamoeba infection. Parasitol Res 116(3):997–1001
Hajaji S, Sifaoui I, López-Arencibia A, Reyes-Batlle M, Valladares B, Pinero JE, Lorenzo-Morales J, Akkari H (2017) Amoebicidal activity of α-bisabolol, the main sesquiterpene in chamomile (Matricariarecutita L.) essential oil against the trophozoite stage of Acanthamoeba castellanii Neff. Acta Parasitol 62(2):290–295
Hegazi AG, El-Fadaly HA, Barakat AM, Abou-El-Doubal SK (2014) In vitro effects of some bee products on T. gondii tachyzoites. Glob Vet 13(6):1043–1050
Hixon KR, Klein RC, Eberlin CT, Linder HR, Ona WJ, Gonzalez H, Sell SA (2019) A critical review and perspective of honey in tissue engineering and clinical wound healing. Adv Wound Care 8(8):403–415
Iqbal K, Iqbal J, Afreen MS (2016) Comparative study on antileishmanial and cytotoxic activity of Lawsonia inermis bark and Aloe vera leaves. Int J Biol Pharm Allied Sci 5(6):1490–1500
Ismail KA, El kadery AAS, Sabry NM, Mohammad OS (2018) A study on the amoebicidal effect of Nigella Sativa aqueous and alcoholic extracts and wheat germ agglutinin on pathogenic Acanthamoeba. J Clin Med Ther 3:12
Kaya Y, Baldemir A, Karaman Ü, Ildız N, Arıcı YK, Kaçmaz G, Kolören Z, Konca Y (2019) Amebicidal effects of fenugreekn (Trigonella foenum-graecum) against Acanthamoeba cysts. Food Sci Nutri 7(2):563–571
Kumar S, Yadav M, Yadav A, Rohilla P, Yadav JP (2017) Anti-plasmodial potential and quantification of aloin and aloe-emodin in Aloe vera collected from different climatic regions of India. BMC Complement Altern Med 17(1):369
Kuźma L, Derda M, Hadaś E, Wysokińska H (2015) Abetane diterpenoids from Salvia sclarea transformed roots as growth inhibitors of pathogenic Acanthamoeba spp. Parasitol Res 114(1):323–327
Liu C, Cui Y, Pi F, Cheng Y, Guo Y, Qian H (2019) Extraction, purification, structural characteristics, biological activities and pharmacological applications of acemannan, a polysaccharide from Aloe vera: a Review. Molecules 24(8):1554
Lorenzo-Morales J, Khan NA, Walochnik J (2015) An Update on Acanthamoeba Keratitis: diagnosis, pathogenesis and treatment. Parasite 22:10
McDonnell G, Russell AD (1999) Antiseptics and disinfectants: activity, action, and resistance. Clin Microbiol Rev 12(1):147–179
Mirzaalizadeh B, Sharif M, Daryani A et al (2018) Effects of Aloe vera and Eucalyptus methanolic extracts on experimental toxoplasmosis in vitro and in vivo. Exp Parasitol 192:6–11
Mohammed SEA, Kabbashi AS, Koko WS, Ansari MJ, Adgaba N, Al-Ghamdi A (2019) In vitro activity of some natural honeys against Entamoeba histolytica and Giardia lamblia trophozoites. Saudi J Biol Sci 26(2):238–243
Nolan VC, Harrison J, Cox JA (2019) Dissecting the antimicrobial composition of honey. Antibiotics 8(4):251
Pegu AJ, Sharma MA (2019) Review on Aloe vera.. IJTSRD Int J Trend Sci Res Dev 3(4):35–40
Saoudi S, Sifaoui I, Chammem N et al (2017) Anti-Acanthamoeba activity of Tunisian Thymus capitatus essential oil and organic extracts. Exp Parasitol 183:231–235
Shamsi H, Shaddel M, Yakhchali M et al (2019) The antileishmanial activity of Aloe vera leaf exudates: in vitro and in vivo. Iran J Dermatol 22(1):18–24
Siddiqui R, Aqeel Y, Khan NA (2016) The development of drugs against Acanthamoeba infections. Antimicrob Agents Chemother 60(11):6441–6450
Sifaoui I, López-Arencibia A, Ticona JC et al (2014) Bioassay guided isolation and identification of anti-Acanthamoeba compounds from Tunisian olive leaf extracts. Exp Parasitol 145:S111–S114
Sinha S, Prakash A, Sehgal R, Medhi B (2018) Comparative effect of manuka honey on anaerobic parasitic protozoans with standard drug therapy under in vitro conditions: a preliminary study. Indian J Pharmacol 50(4):197–203
Skryabina YV, Astakhov YS, Konenkova YS et al (2019) Acanthamoeba keratitis. Review of literature. Case reports. Ophthalmol J 12(1):59–71
Vélez E, Campillo G, Morales G, Hincapié C, Osorio J, Arnache O (2018) Silver nanoparticles obtained by aqueous or ethanolic Aloe vera extracts: an assessment of the antibacterial activity and mercury removal capability. J Nanomater 7215210
Yousuf FA, Mehmood MH, Malik A, Siddiqui R, Khan NA (2016) Antiacanthamoebic properties of natural and marketed honey in Pakistan. Asian Pac J Trop Biomed 6(11):967–972
Acknowledgments
The authors are grateful to Prof. Dr. Gamal Elden Mostafa Mohamed, Botany Department, Faculty of Agriculture, Zagazig University-Egypt, for providing the honey and his help in identification of A. vera and preparation of the extract.
Funding
None.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declared that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Kadry, G.M., Ismail, M.A.M., El-Sayed, N.M. et al. In vitro amoebicidal effect of Aloe vera ethanol extract and honey against Acanthamoeba spp. cysts. J Parasit Dis 45, 159–168 (2021). https://doi.org/10.1007/s12639-020-01292-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12639-020-01292-8