ВИКОРИСТАННЯ ІНТЕГРАЛЬНИХ ІНДЕКСІВ СТРУКТУРИ ЛЕЙКОЦИТАРНОЇ ФОРМУЛИ ДЛЯ ОЦІНКИ СТУПЕНЯ НАПРУЖЕННЯ АДАПТАЦІЙНИХ ПРОЦЕСІВ PELOPHYLAX RIDIBUNDUS (AMPHIBIA: RANIDAE) В УМОВАХ ПРИРОДНОЇ ГЕМОПАРАЗИТАРНОЇ ІНВАЗІЇ
Journal Title: Вісник Запорізького національного університету. Біологічні науки - Year 2016, Vol 1, Issue 1
Abstract
It is known, that adaptative processes stress in an organism can occur under external and internal factors influence. Haemoparasitic invasion can be one of internal factors which are capable to break homeostasis in organism. Haemoparasites are a big enough group of unicells with a wide areal, owners variety, various pathogenicity level and so forth. There is a little-studied and consequently opened question about connection between haemoparasitic loading and adaptative processes stress in green frog organism. The material was gathered in Hortitsa island territory (Zaporozhye). The research objects were lake frogs Pelophylax ridibundus (Pallas, 1771), samples volume was 52 individuals. Researches were spent in laboratory conditions on the basis of biological faculty of Zaporozhian national university. All researches were spent with observance of the principles stated in the Convention on vertebrate animals protection (Strasbourg, France, 1986). Individuals distribution to groups was occurred after haemoparasites presence definition in blood films. Haemoparasites identification was spent by morphometric indices. Hematological researches (leukocytes determination, leucogram calculation) were made with the help of standard methods. Blood films were made from 0,01 ml of blood, parasitic cargo analysis was estimated on invasion extensiveness indexes (EI,% ) and invasion intensity (II, examples). Endoglobular parasite invasion intensity was defined with their calculation on 1000 erythrocytes. Extracellular haemoparasites invasion intensity was counted on blood film (0,02 ml). Integral leucocytal index was counted for organism nonspecific resistance estimation: the ratio of neutrophils percentage sum to lymphocytes percentage quantity (N/L). K.Shennon’s entropy was used for an estimation of blood leucogram absolute entropy (LAE). «The relative system organization» was used for domination index (R) definition. The obtained quantitative data are processed statistically by means of application computer programs package «Statistica 6.0». Distinctions reliability was estimated on Mann-Whitney’s nonparametric test size. The indices difference was considered statistically significant at 95% and more significance value (p ≤ 0,05). In lake frogs P. ridibundus blood unicell parasites, which were referred to 3 taxonomic genera, have been found out: endocellular – Hepatozoon Miller 1908 (Apicomplexa: Adeleorina), Dactylosoma Labbе, 1894 (Apicomplexa: Dactylosomatidae) and extracellular – Trypanosoma Gruby 1843 (Kinetoplastida: Trypanosomatidae), and also nematodes larval stages – microfilaria – from subseries Filariata. Revealed haemoparasites specific definition is proceeding. In considered sample individuals, which were free from haemoparasites, and individuals with monoinvasion were not revealed, EI with adeleid coccidias Hepatozoon (Hp), and also Trypanosoma (Tr) has made 100% , Dactylosoma EI (Dc) – 13,04% , microfilaria EI (Mc) – 43, 48% . Average Hepatozoon II in the sample was 13,32 ± 3,44 examples on 1000 erythrocytes, II Dactylosoma – 0,38 ± 0,31 examples on 1000 erythrocytes, II Trypanosoma – 58,78 ± 10,94 examples on blood film (0,01 ml) and II microfilaria – 472,39 ± 296,46 examples on blood film (0,01 ml). By carried out research results it has been established, that in blood of all of P. ridibundus individuals with a various haemoparasitic invasion combination lymphocytes has made the biggest cell pool, that means that blood has lymphocyte profile. Green frogs lymphocytes, similarly to the higher vertebrate lymphocytes, are the central immune system component, providing all basic immunity reactions, in particular adaptive immunity formation. Lymphocytes relative quantity reduction at individuals with poliinvasion (Hp+Tr+Mc+Dc) on 13,62% (р < 0,01) in comparison with the individuals, having threefold invasion (Hp+Tr+Mc) is established. Leukocytes granulocytic pool changes analysis in P. ridibundus peripheral blood testifies to segmentated neutrophils relative quantity significant increase on 23,87% (р < 0,01) and stab neutrophils significant increase on 30,71% (р < 0,01) at individuals with poliinvasion in comparison with individuals with threefold invasion. Absolute quantity in different leukocytes pools changes analysis allows to ascertain authentic increase in all granulocytes forms at individuals with poliinvasion (Hp+Tr+Mc+Dc) in comparison with individuals with threefold (Hp+Tr+Mc) and double (Hp+Tr) invasion. Among P. ridibundus investigated groups with different haemoparasitic loading degree the greatest stress index was defined in individuals with combined poliinvasion (Hp+Tr+Mc+Dc), that on 14,21% (р < 0,01) and on 11,58% (р < 0,01) is more than at individuals with double (Hp+Tr) and threefold (Hp+Tr+Mc) invasion accordingly. By carried out research results the LAE changes and domination index in P. ridibundus with different haemoparasitic loading degree have found out their multidirectional changes, which testify to system structural organization reduction (its disorganization increase), at the expense of relative organization reduction in group of individuals with poliinvasion (Hp+Tr+Mc+Dc) unlike two other groups. The obtained data have shown possibility of integral leucocytal indices use for P. ridibundus adaptative processes stress degree estimation in natural haemoparasitic invasion conditions.
Authors and Affiliations
В. Ю. Задорожня
Keywords
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