Staining of Parasitic Helminths by Extracts of Allium cepa, Ju-glans regia, and Rubia tinctorum: An Approach to Herbal Dyes
AbstractBackground: Although carmine, as a synthetic dye, is the major substance for staining helminths, it may impose some adverse effects on human health. In the present study, we evaluated the aqueous extracts of onion (Allium cepa) skin, walnut (Juglans regia) husk, and madder (Rubia tinctorum) roots as potential herbal dyes for staining parasitic helminths.Methods: Aqueous solutions (5%, 10% and 20%, w/v) of each herbal prepared from dried and powdered husk of walnut, skins of red onion, and madder roots in distilled water. Parasitic helminths including Fasciola spp., Dicrocoelium denderiticum, Echinococcus gronulosus protoscolices, Moniezia spp., and Haemonchus contortus were stained by different concentrations of herbal dyes according to carmine staining method. The structural clarity and quality of stained internal organs of the helminths such as suckers, intestine, and reproductive systems were scored by semi-quantitative evaluation in comparison with carmine stained samples.Results: The optimum concentrations of extracts for helminths staining were 10% (w/v) of A. cepa and J. regia, and 20% (w/v) of R. tinctorum with final scores of 3.1, 3 and 2.8, respectively. In general, A. cepa and J. regia extracts showed higher quality in staining Platyhelminthes, while R. tinctorum extract presented relatively higher quality in staining Nematoda.Conclusion: Considering proper quality of A. cepa, J. regia and R. tinctorum extracts in staining the helminths, they may be safe, eco-friendly, and inexpensive alternatives to carmine dye.
Muller R. Worms and human disease. 2nd ed. London: CABI Publishing; 2002.
Greenhawt MJ, Baldwin JL. Carmine dye and cochineal extract: hidden allergens no more. Ann Allergy Asthma Immunol. 2009; 103(1):73-5.
Tabar-Purroy AI, Alvarez-Puebla MJ, Ac-ero-Sainz S, García-Figueroa BE, Eche-chipía-Madoz S, Olaguibel-Rivera JM, Quirce-Gancedo S. Carmine (E-120)–induced occupational asthma revisited. J Allergy Clin Immunol. 2003; 111(2):415-9.
Afshar VJ. The process and methods of dyeing fibers with natural dyes. 2nd ed. Tehran: Art University Press; 2002.
Bhuyan R, Saikia CN. Isolation of colour components from native dye-bearing herbals in northeastern India. Bioresour Technol. 2005; 96(3):363-72.
Daryani A, Sharif M, Meigouni M. Staining of Fasciola hepatica by natural herbal dyes. Comp Clin Pathol. 2011; 20(4): 305-8.
Kumar N, Mehul J, Das B, Solanki JB. Staining of Platyhelminthes by herbal dyes: An eco-friendly technique for the taxono-mist. Vet World. 2015; 8(11):1321-5.
Cheng CW, Saad SMd, Abdullah R. Alter-native staining using extracts of hibiscus (Hibiscus rosa-sinensis L.) and red beet (Beta vulgaris L.) in diagnosing ova of intestinal nematodes (Trichuris trichiura and Ascaris lumbricoides). Eur J Biotechnol Biosci. 2014; 1(5): 14-8.
Sharma A, Grover E. Colour fastness of walnut dye on cotton. Indian J Nat Prod Resour. 2011; 2(2): 164-9.
Tutak M, Benli H. Colour and fastness of fabrics dyed with walnut (Juglans regia L.) base natural dyes. Asian J Chem. 2011; 23(2): 566-8.
Vankar PS, Shanker R, Wijayapala S. Dye-ing of cotton, wool and silk with extract of Allium cepa. Pigm Resin Technol. 2009; 38(4): 242–47.
Silva AB, Silva MG, Arroyo PA, Barros MASD. Dyeing mechanism of wool and silk with extract of Allium Cepa. Chem Eng Trans. 2013; 32: 715-20. doi: 10.3303/CET1332120
Ghavami MB, Rahimi P, Haniloo A, Mo-savinasab SN. Genotypic and phenotypic analysis of Fasciola isolates. Iran J Parasitol. 2009; 4(3): 61-70.
Farhadi M, Fazaeli A, Haniloo A. Genetic characterization of livestock and human hydatid cyst isolates from northwest Iran, using the mitochondrial cox1 gene se-quence. Parasitol Res. 2015; 114(12):4363-70.
Al-Amura MF, Hassen ZA, Al-Mhanawi BH. Staining technique for helminth para-sites by use red beet (Beta vulgaris L.) extract. Bas J Vet Res. 2012; 11(1): 283-92.
Chandravanshi S, Upadhyay SK. Interac-tion of natural dye (Allium cepa) with ionic surfactants. J Chem. 2013; 6 pages. doi: 10.1155/2013/685679
Uddin MG. Effects of different mordants on silk fabric dyed with onion outer skin extracts. J Textil. 2014; 8 pages. doi: 10.1155/2014/405626
Bechtold T. Natural colorants in hair dye-ing. In: Bechtold T, Mussak R, editors. Handbook of natural colorants. New York: John Wiley & Sons; 2009. P. 339-350.
Mirjalili M, Karimi L. Extraction and char-acterization of natural dye from green wal-nut shells and its use in dyeing polyamide: focus on antibacterial properties. J Chem. 2013; doi: 10.1155/2013:375352.