Hematological and cytochemical parameters of peripheral blood of Wistar Hannover rats after exposure to epoxiconazole
Keywords:epoxiconazole, hematotoxicity, anemia, succinate dehydrogenase, acid phosphatase, naphthol-AS-D-chloroacetateterase, macrophages, "tailed" lymphocytes.
Introduction. Epoxiconazole is a highly effective triazole fungicide. It is a part of many modern compounds for the control of phytopathogens. Widely used in agriculture in all over the world and in Ukraine, in particular. Despite its hepatotoxic characteristics, this fungicide alters the hematological parameters of blood.
The aim of the study – to investigate the effects of generic triazole fungicide epoxiconazole, 95% on hematological and cytochemical parameters of peripheral blood of Wistar Hannover rats in acute experiment.
Research Methods. 10 healthy males of Wistar Han rats were equally divided into control (0 mg/kg/bw) and experimental groups. Dose 1580 mg/kg/bw of epoxiconazole (1/2 LD50) was administrated once orally by gavage to 5 experimental rats. Peripheral blood was studied at 0 and 1, 3, 7, 14 day after exposure (DAE). RBC, HGB, HCT, erytrocyte indices MCV, MCH, MCHC, WBC and PLT were, hemogram and morphological disturbances of cells were studied,percentage ratio of different types of leukocytes was calculated.The cytochemical status of leukocytes was assessed on the basis of determining the enzymatic activity of specific naphthol-AS-D-chloroacetateterase in neutrophils, succinate dehydrogenase and acid phosphatase in lymphocytes.
Results and Discussion. Erythrocytosis in response to pesticide intoxication with a decrease of mean cellhemoglobin concentration in one erythrocyte; the reduction of hemoglobin concentration in the end of experiment against the background of active polychromasia confirmed the anemic effect of epoxiconazole. Reactive neutrophilia, stimulation of lymphocytopoiesis and monocytopoiesis, appearance of macrophages in the peripheral blood, increased activity of naphthol-AS-D-chloroacetateterase, succinate dehydrogenase and acid phosphatase were considered as compensatory mechanisms. Reverse quantitative changes in the ratio of lymphocytes subpopulations based on the activity of acid phosphatase were noted.
Conclusions. Results of the study of generic fungicide epoxiconazole, 95% hematotoxic activity in the acute experiment on Wistar Hannover male rats showed the development of latent anemia, which was manifested in the late-term study; activation of monocytopoesis; increase of macrophages in the peripheral blood; changes in the immune response of the body and the appearance of atypical normocytes and lymphocytes with nuclear abnormalities.
Kapliuchenko, A.H. (2016). Praktychne znachennia ta zastosuvannia pokhіdnykh 1,2,4-trіazolu [Practical value and application of derivatives of 1,2,4-triazole]. Zaporizhzhia: ZDMU [in Ukrainian].
(2015). Peer review of the pesticide risk assessment of the active substance epoxiconazole. EFSA Journal, 13 (6), 4123.
Chambers, J.E. (2014). Human and ecological risk assessment of a crop protection chemical: a case study with the azole fungicide epoxiconazole. Critical Reviews in Toxicology, 44, 2, 176-210.
Schmidt, F. (2016). Effects of combination azole fungicides in male rats in a broad dose range. Toxicology, 355, 54-63
EPA MEMORANDUM January 24, 2001 Epoxiconazole – Report of the Cancer Assessment Review Committee, USA, 42.
Guide for the care and use of laboratory animals. LAR Publication, National Academy Press, USA, 1996, 22.
OECD Principles of Good Laboratory Practice. ENV/MC/CHEM(98)17. Environment Directorate Organisation for Economic Cooperation and Development, Paris, 1998.
Prodanchuk, M.G. (2015). Stvorennia іstorychnoho kontroliu hematolohіchnykh pokaznykіv peryferychnoi krovі shchhurіv Wistar Han [Create a historical monitoring of hematological parameters of peripheral blood of Wistar rats, Han]. Suchasni problem toksykolohii, kharchovoi ta
khimichnoi bezpeky – Modern Problems of Toxicology, Food and Chemical Safety, 4 (72), 35-40 [in Ukrainian].
US EPA – Pesticides – Fact Sheet for Epoxiconazole, United States Office of Prevention, Pesticides Environmental Protection and Toxic Substances Agency (7505P), August 2006.
Shulyak, V.G. (2002). Dostizheniya v oblasti izucheniya vliyaniya pestitsidov na sistemu krovetvoreniyaa [Advances in the study of the impact of pesticides on the hematopoietic system]. Suchasni problem
toksykolohii – Modern Problems of Toxicology, 1, 42 [in Russian].
Novikova, S. (2017). Klinicheskaya i laboratornaya gematologiya [Clinical and laboratory рaematology]. Retrieved from: http://www.clinlab.info/Hemocytology/ Reticulocytes-counting-49 [in Russian].
Menshikov, V.V. (1987). Laboratornye metody issledovaniya v klinike [Laboratory methods in the clinic]. Moscow: Meditsyna [in Russian].
Butenko, Z.A. (1974). Tsitokhimiya i elektronnaya mikroskopiya kletok krovi i krovetvornykh organov [Cytochemistry and electron microscopy of blood cells and blood-forming organs]. Kyiv: Naukova Dumka [in
Havrylov, A. K. (1987). Depresii krovotvorennia [The Depression of hematopoiesis]. Moscow: Meditsyna [in Russian].
Kravtsov, V. (2017). Nuclear abnormalities of lymphocytes as markers for the simplest bioindication test in case of mass casualty events involving radiation exposure. Emergency Medicine (Los Angel), 7, 356.
Çayır, A. (2014). Micronuclei, nucleoplasmic bridges, and nuclear buds induced in human lymphocytes by the fungicide signum and its active ingredients (boscalid and pyraclostrobin). Environmental Toxicology,
, 7, 723-732
Zeljezic, D. (2004). Chromosomal aberrations, micronuclei and nuclear buds induced in human lymphocytes by 2, 4-dichlorophenoxyacetic acid pesticide formulation. Toxicology, 200, 1, 39-47
Efimtseva, E.A. (2007). Aktivnost esteraz v tkanyakh razlichnykh otdelov zheludochno-kishechnogo trakta severnogo olenya [The Activity of esterases in different tissues of the gastrointestinal tract of eindeer]. Selskokhozyaystvennaya biologiya – Agricultural Biology, 6, 77-80 [in Russian].
Khuneryakova, N.V. (2008). Razrabotka metoda opredeleniya aktivnosti suktsinatdegidrogenazy limfotsitov kak pokazatelya adrenergicheskoy regulyatsii v organizme [Development of a method for determining the
activity of succinate dehydrogenase in lymphocytes as an indicator of adrenergic regulation in the body]. Candidate’s Extended abstract. Pushchino [in Russian].
Radchenko, O. M. (2009). Klіtinnyi іmunіtet za umov rіznykh typіv adaptatsіinykh reaktsii [Cellular immunity in conditions of various types of adaptive reactions]. Medychna hidrolohiia ta reabilitatsiia – Medical
Hydrology and Rehabilitation, 7, 3, 57-60 [in Ukrainian].
Hnidoi, I.M., & Datarock, I. (2002). Іmunnyi status u dіtei u razі dіi svyntsiu v nyzkyhh dozah [Immune status in children with lead exposure in low doses]. Ukrainskyi med. chasopys – Ukrainian Medical Journal, 6 (32), XIXII, 125-127 [in Ukrainian].
Prykhodko, E.A. (2009). Morfofunktsіonalnі zmіny peryferychnoi krovі v umovakh dіi ekzohennykh chynnykіv khіmіchnoї prirodi [Morpho-functional changes of the peripheral blood under the action of exogenous chemical
factors]. Visnyk Sumskoho derzhanoho un-tu – Bulletin of Sumy State University, 2, 1, 34-42 [in Ukrainian].
Vorobel, A.V. Osnovy hematolohіi [Fundamentals of Hematology]. Retrieved from: http://22.214.171.124/ elib/local/349.pdf