Identification of Fasciola Species Using Tegumental Spines in Tissue Sections
-
Arezoo FADAVI
,
Kayhan ASHRAFI
,
Hamid HASSANPOUR
,
Mohamad Bagher ROKNI
,
Seyyed Mostafa HOSSEINI
,
Arezoo BOZORGOMID
,
Leila HOSSEINPOUR
,
Faezeh NAJAFI
,
Gholamreza MOWLAVI
Abstract
Background: Efforts to find a reliable non-molecular means of identification has been the main purpose of the current work that always is persuaded by researchers interested in the field of parasitology.
Methods: Adult fasciolids were obtained from the slaughterhouses in different parts of Iran in 2017, and investigated using the classical old fashion morphological appearances of the worms implementing a camera lucida equipped microscope. Histological procedure was subsequently performed for almost the entire collected adult worms followed by Hematoxylin and Eosin (H&E) staining technique. DNA extraction and RFLP-PCR technique were carried out for the entire fasciolid liver flukes. To attain more comparable morphological conclusions, Scanning Electron Micrographs were also implemented for two molecularly identified fasciolids.
Results: Based on spine morphology observed in worm’s tissue sections two types of tegumental spines, “pointed” and “molar” shapes have been identified addressing to distinguish F. hepatica and F. gigantica species respectively. The present identification has been also supported by Molecular analysis using RFLP-PCR technique.
Conclusion: There are some hidden morphological characters implemented in species identification for certain helminths. Meanwhile, the emergence of computer image analysis system (CIAS) on the scene of taxonomy, has revolutionized the accuracy of measurement in morphology by employing detailed parameters that have not been regarded before. The current study has illustrated the tegumental spines of two Fasciola species in tissue sections which has not been enough considered in helminthological publications so far.
2. Mas-Coma MS, Esteban J, Bargues M (1999). Epidemiology of human fascio-liasis: a review and proposed new classifi-cation. Bull World Health Organ, 77(4):340-6.
3. Robert W, Tolan JR (2011). Fascioliasis due to Fasciola hepatica and Fasciola gigantica in-fection: an update on this ‘neglect-ed’neglected tropical disease. Lab Medicine, 42 (2): 107-116.
4. Beesley NJ, Caminade C, Charlier J, et al (2018). Fasciola and fasciolosis in rumi-nants in Europe: Identifying research needs. Transbound Emerg Dis, 65: 199-216.
5. Mas‐Coma S, Valero MA, Bargues MD (2009). Chapter 2. Fasciola, lymnaeids and human fascioliasis, with a global overview on disease transmission, epi-demiology, evolutionary genetics, molecu-lar epidemiology and control. Adv Parasi-tol, 69: 41-146.
6. Afshan K, Valero M, Qayyum M et al (2014). Phenotypes of intermediate forms of Fasciola hepatica and Fasciola gigantica in buffaloes from Central Punjab, Pakistan. J Helminthol, 88 (4): 417-426.
7. Periago M, Valero M, Panova M et al (2006). Phenotypic comparison of allopatric populations of Fasciola hepatica and Fasciola gigantica from European and African bovines using a computer image analysis system (CIAS). Parasitol Res, 99 (4): 368-378.
8. Mowlavi G, Salehi M, Eshraghian MR et al (2012). Morphometric differentiation be-tween camel and sheep strains of Echi-nococcus granulosus using computer im-age analysis system (CIAS). Asian Pac J Trop Med, 5 (1): 58-61.
9. Itagaki T, Tsutsumi K-I, Ito K et al (1998). Taxonomic status of the Japanese triploid forms of Fasciola: comparison of mito-chondrial ND1 and COI sequences with Fasciola hepatica and Fasciola gigantica. J Parasitol, 84(2): 445-8.
10. Ashrafi K, Valero M, Panova M et al (2006). Phenotypic analysis of adults of Fasciola hepatica, Fasciola gigantica and interme-diate forms from the endemic region of Gilan, Iran. Parasitol Int, 55 (4): 249-260.
11. Marcilla A, Bargues M, Mas-Coma S (2002). A PCR-RFLP assay for the distinction between Fasciola hepatica and Fasciola gigantica. Mol Cell Probes, 16 (5): 327-333.
12. Itagaki T, Kikawa M, Terasaki K et al (2005). Molecular characterization of partheno-genic Fasciola sp. in Korea on the basis of DNA sequences of ribosomal ITS1 and mitochondrial NDI gene. J Vet Med Sci, 67 (11): 1115-8.
13. Ichikawa M, Itagaki T (2010). Discrimination of the ITS1 types of Fasciola spp. based on a PCR–RFLP method. Parasitol Res, 106 (3): 757-61.
14. Ashrafi K (2015). The status of human and animal fascioliasis in Iran: A narrative re-view article. Iran J Parasitol, 10 (3): 306-28.
15. Olsen A, Frankena K, Toft N et al (2015). Prevalence, risk factors and spatial analy-sis of liver fluke infections in Danish cat-tle herds. Parasit Vectors, 8 (1): 160.
16. Soulsby EJL (1982). Helminths, arthropods and protozoa of domesticated animals. 7th ed. Ballierre Tindall. London. pp 40.
17. Chaudhry U, van Paridon B, Shabbir MZ et al (2016). Molecular evidence shows that the liver fluke Fasciola gigantica is the predominant Fasciola species in rumi-nants from Pakistan. J Helminthol, 90 (2): 206-213.
18. Mas-Coma S, Bargues M, Valero M (2018). Human fascioliasis infection sources, their diversity, incidence factors, analytical methods and prevention measures. Para-sitology, 145 (13): 1665-1699.
19. Meyers WM, Neafie RC, Marty AM et al (2000). Pathology of infectious diseases. 1st ed. Amer Registry of Pathology. Washington. pp 80-81.
20. Dangprasert T, Khawsuk W, Meepool A et al (2001). Fasciola gigantica: surface to-pography of the adult tegument. J Helmin-thol, 75 (1): 43-50.
21. Srimuzipo P, Komalamisra C, Choochote W et al (2000). Comparative morphometry, morphology of egg and adult surface to-pography under light and scanning elec-tron microscopies, and metaphase karyo-type among three size-races of Fasciola gigantica in Thailand. Southeast Asian J Trop Med Public Health, 31 (2): 366-73.
| Files | ||
| Issue | Vol 49 No 4 (2020) | |
| Section | Original Article(s) | |
| Keywords | ||
| Fasciola Tissue section Diagnosis Tegumantal spine | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
