ORIGINAL_ARTICLE Anticoagulant resistance in Synanthropic rodents in the Stara Zagora region, Bulgaria The anticoagulant rodenticides are the most commonly used toxicants to control rodents nowadays. Therefore, developing resistance to them is an issue of great importance for pest control. The aim of this study was to investigate the sensitivity of synanthropic rodents in the Stara Zagora region, Bulgaria to some of the most significant first (warfarin and coumatetralyl) and second (bromadiolone and brodifacoum) generation anticoagulants. Resistance tests were carried out by a standard protocol using lethal feeding period tests and blood clotting response tests according to the European and Mediterranean Plant Protection Organization (Paris, France) standard. Studies were performed on 278 wild synanthropic rodents – 67 house mice (Mus musculus), 153 roof rats (Rattus rattus) and 58 brown rats (Rattus norvegicus). The rodents belonged to 11 populations inhabiting 9 animal farms in the region of Stara Zagora, Southern Bulgaria. High-level resistance to warfarinwas established in 100% of surveyed house mice and 92.1% of roof rats. Resistance to coumatetralyl was registered in 62.5% of the tested roof rats. Low-level resistance to bromadiolone was found in 38.5% of the surveyed roof rats and 23.1% of house mice. There was no resistance registered in brown rats. The sensitivity of all three rodent species to the strategic anticoagulant brodifacoum was high, and there were no signs of resistance. The results proved the resistance among synanthropic rodents and led to the conclusion that the resistance in house mice and roof rats to warfarin and coumatetralyl tends to be the main issue in pest control. https://macvetrev.mk/Files/Article/2020/10.2478/macvetrev-2019-0010/macvetrev-2019-0010.pdf 2019-03-15T09:00:00 61 70 10.2478/macvetrev-2019-0010 resistance anticoagulant rodenticides synantropic rodents Georgi G. Zhelev zhelev_vet@abv.bg false 1 Department of Epidemiology, Infectious Diseases and Preventive Medicine, Faculty of Veterinary Medicine, Trakian University, Stara Zagora 6000, Bulgaria LEAD_AUTHOR Kojcho P. Koev false 2 Department of Epidemiology, Infectious Diseases and Preventive Medicine, Faculty of Veterinary Medicine, Trakian University, Stara Zagora 6000, Bulgaria AUTHOR Vladimir D. Dimitrov false 3 Veterinarian, Varna 9000, Bulgaria AUTHOR Vladimir S. Petrov false 4 Department of Epidemiology, Infectious Diseases and Preventive Medicine, Faculty of Veterinary Medicine, Trakian University, Stara Zagora 6000, Bulgaria AUTHOR Arsov, R., Yovchev, E., Girginov, G., Dimov, Iv. (1988). Epizootology. In:E. Yovchev (Ed.), Deratization, third part (pp. 289-309), Sofia:Zemizdat. 1 Tasheva, M., Kontev, Hr. (2000). Guide to controlling harmful rodents (80 p.), Sofia:Videnov&son. 2 Karadzhov, S., Ivanov, Y., Nedelchev, N., Kesiakova, S. (2004). Disinfection, disinsection and deratization in foot-and-mouth disease. In:S. Kesiakova (Ed.), Deratization (pp. 133-157), Sofia:Milena-print. 3 Michailova, A., Georgieva, Tz., Dimitrova N. (1998). List of disinfection, disinsection and deratization preparations authorized in the Republic of Bulgaria. In:Deratization preparations (pp. 181-189), Sofia:Videnov&son. 4 Hadlеr, M., Buckle, Al. (1992). Forty five years of anticoagulant rodenticides –past, present and future trends. Proceeding of the 15th Vertebrate Pest Conference. March, 3-5, (pp. 149-155), Hyatt Newporter, Newport Beach, California, USA. 5 Ishizuka, M., Tanikawa, Ts., Tanaka, K.D., Heewon, M., Okajiama, F., Sakamoto, K.Q., Fujita, Sh. (2008). Pesticide resistance in wild mammals-Mechanisms of anticoagulant resistance in wild rodents. J. Toxicol. Sci. 33(3):283-291. PMid:18670159 6 10.2131/jts.33.283 Andru, J., Cosson, J. F., Caliman, J. P., Benoit, E. (2013). Coumatetralyl resistance of Rattus tanezumi infesting oil palm plantations in Indonesia. Ecotoxicology 22, 377–386. PMid:23264020 7 10.1007/s10646-012-1032-y Corrigan, R.M. (2001). Rodent control - a practical guide for pest management professionals. (353 pp.). Cleveland:GIE Media. PMCid:PMC171≁ 8 Zhelev, G., Kesyakova, Sl. (2012). Building resistance to anticoagulant rodenticides in deratization practice. Veterinary Collection (BG) 1, 42-45. 9 Boyle, C.M. (1960). Case of apparent resistance of Rattus norvegicus Berkenhout to anticoagulant poisons. Nature 188, 517-525. 10 10.1038/188517a0 Lund, M. (1984). Resistance to second-generation anticoagulant rodenticides. Proceeding of the 11th Vertebrate Pest Conference. March, 6-8, (pp. 89-94). Sacramento, California, USA. 11 Myllymäki, A. (1995). Anticoagulant resistance in Europe:appraisal of the data from the 1992 EPPO questionnaire. Pesticide Sci. 43, 69–72. 12 10.1002/ps.2780430111 Pelz, H.-J. (2007). Spread of resistance to anticoagulant rodenticides in Germany. Int J of Pest Manage 53(4):299-302. 13 10.1080/09670∃01245223 Rodenticide Resistance Action Committee (2016). RRAC guidelines on anticoagulant rodenticide resistance management (32 pp.). Brussels, Belgium. [cited 2017 October 19] http://www.rrac.info/content/uploads/RRAC_Guidelines_Resistance.pdf 14 OEPP/EPPO (1995). Efficacy evaluation of rodenticides:PP 1/198(1) „Testing rodents for resistance to anticoagulant rodenticides“(pp. 65-71). European and Mediterranean Plant Protection Organization, Paris, France. 15 Pelz, H.-J., Klemann, N. (2004). Rat control strategies in organic pig and poultry production with special reference to rodenticide resistance and feeding behavior. Wagening J Life Sci. 52(2):173-184. 16 10.1016/S1573-5214(04)80012-5 Garcia-Manzano, A., Gonzalez-Llaven, J., Lemini, C., Rubio-Poo, C. (2001). Standardization of rat blood clotting tests with reagents used for humans. Proc. West. Pharmacol. Soc. 44, 153-155. PMid:11793967 17 Cowan, D. P., Dunsford, G., Gill, J. E., Jones, A., Kerins, G. M., MacNicoll, A. D., Quy, R. J. (1995). The impact of resistance on the use of second-generation anticoagulants against rats on farms in southern England. Pestic. Sci. 43, 83-93. 18 10.1002/ps.2780430114 Bailey, C.I., Eason, C.T. (2000). Anticoagulant resistance in rodents. Conservation Advistory Science Notes 297, 1-8. 19 Bailey, C.I., Fisher, P., Eason, C.T. (2005). Assessing anticoagulant resistance in rats and coagulation effects in birds using small-volume blood samples. Science for conservation 249, 1-22. 20 Dubock A. C., Kaukeinen, D. E. (1978). Brodifacoum (Talon®rodenticide), a novel concept. Proceeding of the 8th Vertebrate Pest Conference. March, 7-9, (pp. 126-137). Red Lion Motor Inn Sacramento, California, USA. 21 MacNicoll, A.D., Kerins, G. M., Dennis, N. J., Gill, J. E. (1996). The distribution and significance of anticoagulant-resistant Norway rats (Rattus norvegicus) in England and Wales, 1988-1995. Proceeding of the 17th Vertebrate Pest Conference. March, 5-7, (pp. 179-185). Rohnert Park, California, USA. 22 Pelz, H.-J., Rost, S., Müller, E., Esther, A., Ulrich, R., Müller, C. (2012). Distribution and frequency of VKORC1 sequence variants conferring resistance to anticoagulants in Mus musculus. Pest Manag Sci. 68(2):254-259. PMid:2189∵ 23 10.1002/ps.2254 Ashton, A.D., Jakson, W. B. (1982). Anticoagulant resistance in the house mouse in North America. Proceeding of a Conference on:30 August-03 September (pp. 181-188). Hampshire, England. 24 Rodenticide Resistance Action Group (2012). RRAG house mouse resistance guideline. Rodenticide Resistance Action Group (11 pp.). Derby, UK. [cited 2017 October 19] http://www.pestmagazine.co.uk/media/244094/rrag-house-mouse-resistance-guidelines-aug-2012.pdf 25 Graves, J.H., Rennison, B.D., Redfern, R. (1976). Resistance of ship rats Rattus rattus L. to warfarin. J Stored Prod Res. 12(2):65-70. 26 10.1016/0022-474X(76)90025-4 Tanaka, K. D., Kawai, Y. K., Ikenaka, Y., Harunari, T., Tanikawa, T., Ando, S., et al. (2012). The genetic mechanisms of warfarin resistance in Rattus rattus found in the wild in Japan. Pestic Biochem Physiol. 103(2):144-151. 27 10.1016/j.pestbp.2012.04.011 Jackson, W. B., Ashton, A. D. (1986). Case histories of anticoagulant resistance. In:National Research Council (Ed.), Pesticide Resistance, In:National Academy Press (Ed.), Strategies and tactics for management (pp. 355–369). Washington, DC:National Academy Press. 28 Tanikawa, T., Harunari, T., Nagaoka, K. (2006). Warfarin-resistant brown rats, Rattus norvegicus, in Aomori Prefecture:the first case reported in Japan. Medical Entomology and Zoology 57(3):241-244. 29 10.7601/mez.57.241 Negrin, O., Borroto, R., del.Solar, S. (1989). Negrin, O., Borroto, R., del Solar, S. (1989). Susceptibility of Mus musculus (Rodentia:Muridae) to warfarin. Science and Technology in Agriculture. Plant Protection (E) 12(3):7-14. [in Spanish] 30 Endepols, S., Schuster, W. (1991). Laboratory investigations of resistance to warfarin of roof rats (Rattus rattus) in North East Germany. Zeitschrift Fuer Angewandte Zoologie 78(4):489-495. 31 Berny, P. (2011). Challenges of anticoagulant rodenticides:resistance and ecotoxicology. Chapter 20. In:Margarita Stoytcheva (Ed.), Pesticides in a modern world- pest control and pesticides exposure and toxicity assessment (pp. 441-468). Croatia:InTech. 32 10.5772/19916 Whisson, D. (1996). Rodenticides for control of norway rats, roof rats and house mice. Poultry fact sheet №23. Davis, California, USA:University of California. 33 Buckle, A. P., Klemann, N., Prescott, C. V. (2012). Brodifacoum is effective against Norway Rats (Rattus norvegicus) in a tyrosine139cysteine focus of anticoagulant resistance, Westphalia, Germany. Pest Manag. Sci. 68(12):1579-1585. PMid:23007948 34 10.1002/ps.3352 Kerins, G., Langton, S.R., Mac Nicoll, A. (1993). The development of a blood clotting response test for discriminating between difenacoum resistant and susceptible Norway rats. Comp Biochem Physiol 104(1):29-36. 35 Rodenticide Resistance Action Committee of CropLife International (2003). A reappraisal of blood clotting response tests for anticoagulant resistance and a proposal for standardized BCR test methods. CropLife International Technical Monograph (15 pp.). Brussels, Belgium. [cited 2017 October 19] http://www.rrac.info/content/uploads/technical_monograph_2003_BCR.pdf 36 Baert, K., Stuyck, J., Breyne, P., Maes, D., Casaer, J. (2012). Distribution of anticoagulant resistance in the brown rat in Belgium. Belg. J. Zool. 142(1):39-48. 37