Abstract
Fasciolosis is an important zoonotic disease caused by the trematode Fasciola hepatica, and it causes great losses in bovine production. The anthelmintic resistance is a major problem in the control of fasciolosis. In this study, the F. hepatica egg development and hatching test (EDHT) was used for the evaluation of the ovicidal activity of commercial drugs, commonly used for treating infected cattle, which reflects F. hepatica anthelminthic resistance in infected bovines, according to recent literature. Bile samples from F. hepatica naturally parasitized cattle were obtained from slaughterhouses in the cities of Lages and Otacílio Costa, Santa Catarina State, Brazil. The bile was washed, the eggs were recovered, quantified, and distributed in universal collectors, with a minimum of 1,000 eggs per vial. Four commercial drugs were used in this study, containing albendazole sulfoxide (ABDZ), closantel (CSTL), nitroxynil (NTXL), and triclabendazole with fenbendazole (TBZF). The drugs were diluted according to the manufacturer instructions. All drugs, and the respective control, were tested in triplicates, with the quantity of recovered eggs determining the number of drugs to be tested. The vials were incubated for 28 days at 27 °C, and the eggs were classified according to their degree of development under a stereomicroscope. In total, 121 egg samples were analyzed. Two samples were identified as resistant to TBZF. Undetermined resistance/susceptibility has been found in two isolates treated with ABDZ, one treated with NTXL and six treated with TBZF. CSTL did not present ovicidal activity and cannot be used in EDHT. This is the first time that commercial drugs were used in F. hepatica EDHT.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The data that support the findings of this study are available on request from the corresponding author, ALC.
References
Aksoy DY, Kerimoglu U, Oto A, Ergwen S, Arslan S, Unal S, Batman F, Bayraktar Y (2005) Infection with Fasciola hepatica. Clin Microbiol Infect 11(11):859–861. https://doi.org/10.1111/j.1469-0691.2005.01254.x
Alba A, Vázquez AA, Sánchez J, Duval D, Hernández HM, Sabourin E, Vittecoq M, Hurtrez-Boussés S, Gourbal B (2018) Fasciola hepatica-Pseudosuccinea columella interaction: effect of increasing parasite doses, successive exposures and geographical origin on the infection outcome of susceptible and naturally-resistant snails from Cuba. Parasit Vectors 11:559. https://doi.org/10.1186/s13071-018-3155-3
Albuquerque RBF, Pereira SA, Melo SN, Belo VS, Arruda MM, Mazetto D, Figueiredo FB (2022) Spatial distribution analysis of bovine fascioliasis cases recorded in an abattoir in the state of Santa Catarina, Brazil. Cienc Rural 52:3. https://doi.org/10.1590/0103-8478cr20210030
Alvarez L, Moreno G, Moreno L, Ceballos L, Shaw L, Fairweather I, Lanusse C (2009) Comparative assessment of albendazole and triclabendazole ovicidal activity on Fasciola hepatica eggs. Vet Parasitol 164(2–4):211–216. https://doi.org/10.1016/j.vetpar.2009.05.014
Alvarez LI, Valladares MM, Canton C, Lanusse CE, Ceballos L (2020) Testing Albendazole Resistance in Fasciola hepatica. In: Cancela, M., Maggioli, G. (eds) Fasciola hepatica. Methods Mol Biol 2137:213–220. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0475-5_16
Américo L, Padilha MAC, Arruda PM, Drescher G, Moura AB, Chryssafidis AL (2022) Epidemiological survey and confirmation of autochthonous cases of bovine fasciolosis in the Serrana Mesoregion of Santa Catarina, Brazil. Front Vet Sci 9:933462. https://doi.org/10.3389/fvets.2022.933462
Arias-Pacheco C, Lucas JR, Rodríguez A, Córdoba D, Lux-Hoppe EG (2020) Economic impact of the liver condemnation of cattle infected with Fasciola hepatica in the Peruvian Andes. Trop Anim Health Prod 4:1927–1932. https://doi.org/10.1007/s11250-020-02211-y
Babják M, Königová A, Burcáková Ľ, Komáromyová M, Dolinská MU, Várady M (2021a) Assessing the efficacy of albendazole against Fasciola hepatica in naturally infected cattle by in vivo and in vitro methods. Vet Sci 8(11):249–257. https://doi.org/10.3390/vetsci8110249
Babják M, Königová A, Urda Dolinská M, Kupčinskas T, Vadlejch J, von Samson-Himmelstjerna G, Petkevičius S, Várady M (2021b) Does the in vitro egg hatch test predict the failure of benzimidazole treatment in Haemonchus contortus? Parasite 28:62–68. https://doi.org/10.1051/parasite/2021059
Boulard C, Carreras F, Van Gool F (1995) Evaluation of nitroxynil and closantel activity using ELISA and egg counts against Fasciola hepatica in experimentally and naturally infected cattle. Vet Res 26(4):249–255
Campos Júnior DA, de Oliveira PR (2005) In vitro evaluation of acaricides efficiency to Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae) from bovines at the region of Ilhéus, Bahia, Brazil. Ciência Rural 35(6):1386–1392. https://doi.org/10.1590/S0103-84782005000600025
Canevari J, Ceballos L, Sanabria R, Romero J, Olaechea F, Ortiz P, Cabrera M, Gayo V, Fairweather I, Lanusse C, Alvarez L (2014) Testing albendazole resistance in Fasciola hepatica: validation of an egg hatch test with isolates from South America and the United Kingdom. J Helminthol 88(3):286–292. https://doi.org/10.1017/S0022149X13000163
Ceballos L, Canton C, Pruzzo C, Sanabria R, Moreno L, Sanchis J, Suarez G, Ortiz P, Fairweather I, Lanusse C, Alvarez L, Martinez-Valladares M (2019) The egg hatch test: a useful tool for albendazole resistance diagnosis in Fasciola hepatica. Vet Parasitol 271:7–13. https://doi.org/10.1016/j.vetpar.2019.06.001
Chryssafidis AL, Fu Y, De Waal T, Mulcahy G (2015) Standardisation of egg-viability assays for Fasciola hepatica and Calicophoron daubneyi: a tool for evaluating new technologies of parasite control. Vet Parasitol 210(1–2):25–31. https://doi.org/10.1016/j.vetpar.2015.03.005
Coles GC, Stafford KA (2001) Activity of oxyclozanide, nitroxynil, clorsulon and albendazole against adult triclabendazole-resistant Fasciola hepatica. Vet Rec 148(23):723–724. https://doi.org/10.1136/vr.148.23.723
Coles GC, Jackson F, Pomroy WE, Prichard RK, von Samson-Himmelstjerna G, Silvestre A, Taylor MA, Vercruysse J (2006) The detection of anthelmintic resistance in nematodes of veterinary importance. Vet Parasitol 136(3–4):167–185. https://doi.org/10.1016/j.vetpar.2005.11.019
Costa RA, Corbellini LG, Castro-Janer E, Riet-Correa F (2019) Evaluation of losses in carcasses of cattle naturally infected with Fasciola hepatica: effects on weight by age range and on carcass quality parameters. Int J Parasitol 11:867–872. https://doi.org/10.1016/j.ijpara.2019.06.005
Echevarria F (2004) Fasciolose. Rev Bras Parasitol Vet 13:100–102
Fairweather I (2011) Reducing the future threat from (liver) fluke: realistic prospect or quixotic fantasy? Vet Parasitol 180(1–2):133–143. https://doi.org/10.1016/j.vetpar.2011.05.034
Fairweather I, McShane DD, Shaw L, Ellison SE, O’Hagan NT, York EA, Trudgett A, Brennan GP (2012) Development of an egg hatch assay for the diagnosis of triclabendazole resistance in Fasciola hepatica: proof of concept. Vet Parasitol 183(3–4):249–259. https://doi.org/10.1016/j.vetpar.2011.07.023
Fairweather I, Brennan GP, Hanna REB, Robinson MW, Skuce PJ (2020) Drug resistance in liver flukes. Int J Parasitol Drugs Drug Resist 12:39–59. https://doi.org/10.1016/j.ijpddr.2019.11.003
Foreyt WJ (1988) Efficacy of a fenbendazole-triclabendazole combination against Fasciola hepatica and gastrointestinal nematodes in sheep. Vet Parasitol 26(3–4):265–271. https://doi.org/10.1016/0304-4017(88)90095-7
Giehl AL (2021) Desempenho da produção animal – Carne bovina. 41 Síntese Anual da Agricultura de Santa Catarina 2019–2020, (pp 93–105). Florianópolis: Ep
González-Garduño R, Ortiz-Pérez DO, Alegría-Jiménez L, Torres-Chable OM, Cruz-Tamayo AA, Zaragoza-Vera CV (2020) Evaluation of anthelmintic drugs against egg development of rumen flukes recovered from cattle raised in the humid tropics of Mexico. J Helminthol 94:e177. https://doi.org/10.1017/S0022149X20000607
Gordon D, Zadoks R, Skuce P, Sargison N (2012) Confirmation of triclabendazole resistance in liver fluke in the UK. Vet Rec 171(6):159–160. https://doi.org/10.1136/vr.e5381
Han T, Wang M, Zhang G, Han D, Li X, Liu G, Li X, Wang Z (2017) Gastrointestinal nematodes infections and anthelmintic resistance in grazing sheep in the Eastern Inner Mongolia in China. Acta Parasitol 62(4):815–822. https://doi.org/10.1515/ap-2017-0098
Hanna REB, McMahon C, Ellison S, Edgar HW, Kajugu PE, Gordon A, Irwin D, Barley JP, Malone FE, Brennan GP, Fairweather I (2015) Fasciola hepatica: a comparative survey of adult fluke resistance to triclabendazole, itroxinil and closantel on selected upland and lowland sheep farms in Northern Ireland using faecal egg counting, coproantigen ELISA testing and fluke histology. Vet Parasitol 207(1–2):34–43. https://doi.org/10.1016/j.vetpar.2014.11.016
Ico-Gómez R, González-Garduño R, Ortiz-Pérez D, Mosqueda-Gualito JJ, Flores-Santiago EDJ, Sosa-Pérez G, Salazar-Tapia AA (2021) Assessment of anthelmintic effectiveness to control Fasciola hepatica and paramphistome mixed infection in cattle in the humid tropics of Mexico. Parasitology 148(12):1458–1466. https://doi.org/10.1017/S0031182021001153
Kamaludeen J, Graham-Brown J, Stephens N, Miller J, Howell A, Beesley NJ, Hodgkinson J, Learmount J, Williams D (2019) Lack of efficacy of triclabendazole against Fasciola hepatica is present on sheep farms in three regions of England, and Wales. Vet Rec 184(16):502. https://doi.org/10.1136/vr.105209
Kelley JM, Elliott TP, Beddoe T, Anderson G, Skuce P, Spithill TW (2016) Current Threat of Triclabendazole Resistance in Fasciola hepatica. Trends Parasitol 32(6):458–469. https://doi.org/10.1016/j.pt.2016.03.002
Kelley JM, Rathinasamy V, Elliott TP, Rawlin G, Beddoe T, Stevenson MA, Spithill TW (2020) Determination of the prevalence and intensity of Fasciola hepatica infection in dairy cattle from six irrigation regions of Victoria, South-eastern Australia, further identifying significant triclabendazole resistance on three properties. Vet Parasitol 277:109019. https://doi.org/10.1016/j.vetpar.2019.109019
Königová A, Urda Dolinská M, Babják M, von Samson-Himmelstjerna G, Komáromyová M, Várady M (2021) Experimental evidence for the lack of sensitivity of in vivo faecal egg count reduction testing for the detection of early development of benzimidazole resistance. Parasitol Res 120(1):153–159. https://doi.org/10.1007/s00436-020-06965-0
Martínez-Valladares M, del Rosario FM, Fernández-Pato N, Castañón-Ordóñez L, Cordero-Pérez C, Rojo-Vázquez FA (2010) Efficacy of nitroxynil against Fasciola hepatica resistant to triclabendazole in a naturally infected sheep flock. Parasitol Res 107(5):1205–1211. https://doi.org/10.1007/s00436-010-1989-5
McConville M, Brennan GP, Flanagan A, Edgar HW, Hanna RE, McCoy M, Gordon AW, Castillo R, Hernández-Campos A, Fairweather I (2009) An evaluation of the efficacy of compound alpha and triclabendazole against two isolates of Fasciola hepatica. Vet Parasitol 162(1–2):75–88. https://doi.org/10.1016/j.vetpar.2009.02.004
Mehmood K, Zhang H, Sabir AJ, Abbas RZ, Ijaz M, Durrani AZ, Saleem MH, Rehman UM, Iqbal MK, Wang Y, Ahmad HI, Abbas T, Hussain R, Ghori MT, Ali S, Kahn AU, Li J (2017) A review on epidemiology, global prevalence and economical losses of fasciolosis in ruminants. Microb Pathog 109:253–262. https://doi.org/10.1016/j.micpath.2017.06.006
Mooney L, Good B, Hanrahan JP, Mulcahy G, de Waal T (2009) The comparative efficacy of four anthelmintics against a natural acquired Fasciola hepatica infection in hill sheep flock in the west of Ireland. Vet Parasitol 164(2–4):201–205. https://doi.org/10.1016/j.vetpar.2009.05.017
Novobilský A, Höglund J (2015) First report of closantel treatment failure against Fasciola hepatica in cattle. Int J Parasitol Drugs Drug Resist 5(3):172–177. https://doi.org/10.1016/j.ijpddr.2015.07.003
Olaechea F, Lovera V, Larroza M, Raffo F, Cabrera R (2011) Resistance of Fasciola hepatica against triclabendazole in cattle in Patagonia (Argentina). Vet Parasitol 178(3–4):364–366. https://doi.org/10.1016/j.vetpar.2010.12.047
Ortiz P, Scarcella S, Cerna C, Rosales C, Cabrera M, Guzmán M, Lamenza P, Solana H (2013) Resistance of Fasciola hepatica against Triclabendazole in cattle in Cajamarca (Peru): a clinical trial and an in vivo efficacy test in sheep. Vet Parasitol 195(1–2):118–121. https://doi.org/10.1016/j.vetpar.2013.01.001
Potârniche AV, Mickiewicz M, Olah D, Cerbu C, Spînu M, Hari A, Györke A, Moroz A, Czopowicz M, Várady M, Kaba J (2021) First report of anthelmintic resistance in gastrointestinal nematodes in goats in Romania. Animals (Basel) 11(10):2761. https://doi.org/10.3390/ani11102761
Rapic D, Dzakula N, Sakar D, Richards RJ (1988) Comparative efficacy of triclabendazole, nitroxynil and rafoxanide against immature and mature Fasciola hepatica in naturally infected cattle. Vet Rec 122(3):59–62. https://doi.org/10.1136/vr.122.3.59
Robles-Pérez D, Martínez-Pérez JM, Rojo-Vázquez FA, Martínez-Valladares M (2014) Development of an egg hatch assay for the detection of anthelmintic resistance to albendazole in Fasciola hepatica isolated from sheep. Vet Parasitol 203(1–2):217–221. https://doi.org/10.1016/j.vetpar.2013.11.020
Romero J, Villaguala C, Quiroz F, Landaeta-Aqueveque C, Alfaro G, Pérez R (2019) Flukicide efficacy against Fasciola hepatica of Triclabendazole and Nitroxynil in cattle of the central valley of Chile. Rev Bras Parasitol Vet 28(1):164–167. https://doi.org/10.1590/S1984-296120180089
Sargison N (2012) Diagnosis of triclabendazole resistance in Fasciola hepatica. Vet Rec 171(6):151–152. https://doi.org/10.1136/vr.e5357
Silva AEP, da Freitas CC, Dutra LV, Molento MB (2020) Correlation between climate data and land altitude for Fasciola hepatica infection in cattle in Santa Catarina, Brazil. Rev Bras Parasitol Vet 29(3):1–7. https://doi.org/10.1590/S1984-29612020065
Wood IB, Amaral NK, Bairden K, Duncan JL, Kassai T, Malone JB Jr, Pankavich JA, Reinecke RK, Slocombe O, Taylor SM et al (1995) World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) second edition of guidelines for evaluating the efficacy of anthelmintics in ruminants (bovine, ovine, caprine). Vet Parasitol 58(3):181–213. https://doi.org/10.1016/0304-4017(95)00806-2
Zárate-Rendón DA, Briones-Montero A, Huaraca-Oré NA, Veirano GS, Levecke B, Geldhof P (2023) Comparison of the therapeutic efficacy of five anthelmintics against natural Fasciola hepatica infections in dairy cattle from the Mantaro Valley, Peru. Vet Parasitol Reg Stud Rep 38:100827. https://doi.org/10.1016/j.vprsr.2022.100827
Funding
This work was funded by Foundation for Research and Innovation of the State of Santa Catarina (FAPESC, grant number 027/2020 − grant term 793/2021). The authors are thankful for the abattoirs that kindly provided the samples used in this investigation.
Author information
Authors and Affiliations
Contributions
ALC and ABM designed the study. PMA and JM collected the samples. PMA, LA, SGRM, FRL carried out the tests. PMA, LA, FRL and ALC wrote the main manuscript. All authors reviewed the manuscript.
Corresponding author
Ethics declarations
Ethical approval
The protocols used in the present study were approved by the Ethics Committee on the Use of Animals of the Agroveterinary Sciences Center (CEUA-CAV-UDESC), protocol number 6602070619.
Consent to participate and consent to publish
Not applicable in this study.
Competing interests
The authors declare that they have no conflict of interest.
Additional information
Section Editor: David Bruce Conn
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Chryssafidis, A.L., Arruda, P.M., Américo, L. et al. The Fasciola hepatica egg development and hatching test (EDHT) using commercial drugs: a simple protocol for tackling anthelminthic resistance and promoting cattle productivity. Parasitol Res 123, 70 (2024). https://doi.org/10.1007/s00436-023-08074-0
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00436-023-08074-0