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Introduction

The steady existence of population and its relative independence as an unified biological sys-tem depends on its ability to sustain an adaptive pop-ulation homeostasis secured by adaptive biochemi-cal and physiological reactions of the separate indi-viduals integrated on a population level.

Each organism possesses a limited quantity of available energy used for growth, development, reproduction and adaptation and for that reason the ability of the population to regulate and support of dynamic homeostasis is relatively limited and de-pends on the effect of the complicated complex of interacting factors of the environment.

The object of the present paper is to study the joint effect of γ-rays and chlorine organic pes-ticides on some bioenergetics genetic functions of model populations of white mice. As a genetic mark-er we use the gene-enzyme of lactatedehydrogenase (LTH) and we judged for the energetic balance by the AT Phase and respiratory activity of isolation of liver mitochondria from the individuals.

Materials and methods

There have been studied white puberty labo-ratory mice of 18-20 g weight divided into five mod-el groups of 5 mice each –three male and two female. The dominant and subordinate individuals were de-termined in each group by measuring the sodium amital resistance of breathing (the stress level) reg-istered by a respiratory device Kalabuchov model Калабухов, Н. �., (1�51). The animals were irradi- �., (1�51). The animals were irradi-�., (1�51). The animals were irradi-(1�51). The animals were irradi-. The animals were irradi-ated for 3hrs and 30 minutes, for 7hrs and for 14hrs with a source of γ-rays Co with 5 mCi activity and the absorbed doses were 8.25 mSv, 17.5 mSv and 35 mSv. The tested pesticides were put into the ani-mals orally in doses corresponding to LD50 for mice namely heptachlor – 146 mg/kg; hexachlorine – 500

JOINT ACTION OF γ – RAYS AND PESTICIDES ON SOME BIOENERGETIC AND GENETIC FUNCTIONS OF MODEL GROUPS

OF WHITE MICE

Aleksandar DOICHINOV

Shumen University Episkop Konstantin Preslavsky, 115, Universitetska Str., Shumen, 9700, Bulgaria

mg/kg and lindan – 125 mg/kg. Юхтин, Н.Н., (1�73 q). 24 hours after the irradiation with γ-rays and the pesticide introduction the animals were decapitated without narcosis and the isolated liver mitochondria and the LDH isoenzyme spectra of a heart muscle were studied. The liver mitochondria were isolated by Johnson and Lardi method with some slight mod-ifications Johnson, D., Lardy, K., (1�67). The elec-tron transport in the respiratory chain was determined by the oxygen consummation of the mitochondri-al suspension, registered polargraphically by a cov-ered platinum electrode Шольц, К., Островский, Д., (1�75). The AT Phase activity was measured by the growth of the Ph quality in the incubation con-ditions, measured by Lowry Lopes method Lowry, O., Lopes, J., (1�64). The protein content in the test was determined by the method of Lowry Lowry, O., et al. (1�61). The LDH isoforms were separated by electrophoresis in 5% polyakrylamid gel (PAAG) by the system of Ornstein Ornstein, L., (1�64) and Da-vis Davis, B., (1�64) in vertical block.

Results and discussions

It is well-known that the chlororganic pesti-cide in small amounts has a strong effect on the ener-gy functions of the liver mitochondria in vivo and in vitro Каменов, Д., (1�86). However, the mechanism of the impact is different and it can show in block-ing the electron transport in the respiratory chain (e.g. hexachlorine), suppressing the transport of the transport systems (heptachlor) or stimulating the AT Phase activity (lindan).

No disturbance in the bioenergetic functions of the isolated liver mitochondria was observed at a single whole body irradiation of the tested animals with the pointed doses. At the same time, the irradi-ation of mitochondria isolated from normal animals, with suspension in doses about 12.5 mSv for 30 min. causes complete loss of their energy functions. The

Оригинален научен трудOriginal Scientific Article

Joint action of γ – rays and pesticides on some bioenergetic and genetic functions of model...

385Proceedings of the III Congress of Ecologists of Macedonia

mitochondria are characterized by extremely high oxygen consumption similar to the frozen or treat-ed with 2.4-DHF or Triton-X 100, which shows that irradiation in vitro causes destructive changes in the membrane of the mitochondria (Fig.1).

The isoenzyme spectrum of the LDH of a heart muscles normally is a distinctly expressed type i.e. H3M and H4 isoforms prevail working at high oxygen content in the tissues. In our previous in-vestigations Дойчинов, А.и др. (1��4) it has been pointed out that at a single whole body irradiation of the organism with γ –rays in doses 35.0mSv for 14 hours a significant change occurs in the propor-tion of the separate fractions of the LDH to the ad-vantage of the aerobic ones – M3H and M4. Besides, the effect depends on the rank of the individual and its position in the ethological structure of the group. This change is distinct in the dominant individuals in the group and does not show in those in lower position in the ethological structure. These changes in the polymorphism of the LDH correlate regularly with a decrease of the organism’s oxygen consump-

tion highly expressed in the individuals of a high rank. The female individuals which do not partici-pate in the construction of the hierarchical structure of the population but play the role of a buffer mod-erating the aggression between the males react simi-larly to the subordinates and do not change the con-tent of the LDH during the irradiation with γ –rays in the given doses (Fig. 2).Presumably this id due to providing the adaptation of the individuals.

The decrease of the oxygen consumption and the lowering of the oxygen content in the tissues re-duce the effect of the influence of the ionizing irra-diation while the increased amount of the anaerobic isoforms of the LDH ensures a normal energy bal-ance of the organism under these conditions. More-over this makes the individuals of the higher rank more adaptive compared to the other ones.

The used pesticides do not have any effect on the isoenzyme spectrum and the rations of the sepa-rate isoforms of the LDH of the organisms.

The joint effect of the γ –rays and pesticides on the animals do not cause synergetic changes in

Fig. 1. Effect of the different doses γ-rays on succinate-oxidase activity of the liver mitochondria of white mice in vitro and in vivo: 1-control; 2- mitochondria suspension, irradiated in vitro with 60Co in dose 1,25 mSv for 30 min; 3-- mitochondria , isolated 24 hrs after a single whole body irradiation of the animal with 60Co in dose 8,25 mSv, 17,5 mSv and 35,0 mSv. Supplements: mitochondria (MC) – 4,68 mg; 2,4-dinitrophenol (DNP) – 80 mM; succinate (S) – 5 mM

Сл. 1. Ефекти предизвикани од влијанието на различни дози на γ –зраците врз активноста на сукцинат-оксидазата во митохондриите на слезината кај белиот лабораториски стаорец in vitro и in vivo: 1-контрола; 2- митохондријална суспензија, озрачена ин витро со 60 Co во доза од 1,25 mSv za vreme od 30 min; 3 - митохондрија, изолирана 24 часа после еднократно озрачување на целото тело на животното со 60 Co во доза од 8,25 mSv,17,5 mSv и 35,0 mSv. Додатоци: митохондрија (MC) - 4,68 mg; 2,4-динитрофенол (DNP) – 80 mM; сукцинат (S) – 5 mM

Aleksandar DOICHINOV

386 Зборник на трудови од III Конгрес на еколозите од Македонија

the isoenzyme content of the LDH but only such that correspond to the ionizing irradiation. The simulta-neous effect of both factors, however, changes sig-nificantly the energy functions of the mitochondria.

Thus, for example the inhibiting actions of the electron transport of heptachlor and hexachlo-rine on the background of the irradiation of the or-ganisms with γ –rays with the given doses, increas-es significantly and the effect is unstochchastic i.s. the degree of inhibiting increases with the increase of the dose. Besides, the effect is more significant in the dominant animals.

The blocking of electron transport in vitro in

the dominant increase by 25% for heptachlor and 17.5% for hexachlorine, while for the female and male individuals at a lower position in the hierar-chical structure it is just 5% and 3.5% respective-ly (Tab. 1).

In the same way the γ –rays influence also on the effect of lindan, as in given doses they cause an effect in vitro. The last one consists in an increase of the AT Phase activity of the liver mitochondria about twice (Fig. 3).

On the basis of the obtained results we can draw the conclusion that the γ –rays in the given dose shave synergetic effect on the tested pesticides.

Fig. 2. Isoenzymes spectrum of LDH from a heart muscle of a white laboratory mouse under the action of γ-rays or /and/ pesticides depending on the rank position of the individual in the ethological structure; K1 – dominants – controls; K2 – subordinates – controls; E1 – dominants, irradiated with γ-rays (35,0 mSv); E2 – subordinates, irradiated with γ-rays (35,0 mSv); E3 – dominants, cultivated with one of the tested pesticides; E4 – subordinates, cultivated with one of the tested pesticides; E5 – dominants, irradiated with γ-rays and cultivated with pesticides; E6 – subordinates, irradiated with γ-rays and cultivated with pesticides

Сл. 2. Спектар на изоензими од LDH изолирани од срцев мускул на бел лабораториски глушец под влијание на γ- зраци или /и/ пестициди зависно од категоризацијата на единката во етолошката структура: K1 – доминанти – контроли; K2 – субординати – контроли; E1 – доминанти – озрачени со g-зраци (35,0 mSv); E2 – субординати – озрачени со г-зраци (35,0 mSv); E3 – доминанти, култивирани со еден од испитуваните пестициди; E4 – субординати, култивирани со еден од испитуваните пестициди; E5 – доминанти, озрачени со γ-зраци и култивирани со пестициди; E6 – субординати, озрачени со г-зраци и култивирани со пестициди.

Tab. 1. Joint action of γ-rays and pesticides in vivo of the intact liver mitochondria of white mice depending on the rank position of the individual in the ethological structure

Таб. 1. Комбинирано влијание на γ-зраците и пестицидите in vivo на интактна митохондрија од слезината на белиот лабораториски глушец, зависно од рангираноста на единката во етолошката структура

Liver mitochondria of white mice Oxygen consumption µAO2 /min/mgdominants subordinates ♀

MC, controls 0,1�6 0,1�4 O,1�7MC + heptahlor 0,128 (34,6�%) 0,123 (37,24) 0,12� (34,18)MC + heptachlor + γ-rays ( 8,25 mSv) 0,125 (36,22%) 0,10� (44,3�) 0,111 (43,37)MC + heptachlor + γ-rays ( 17,5 mSv) 0,126 (35,71%) 0,0�6 (51,02) 0,0�5 (51,53)MC + heptachlor + γ-rays ( 35,0 mSv) 0,11� (3�,2�%) 0,084 (57,14) 0,0�1 (53,57)MC + hexahloran 0,154 (21,43%) 0,150 (23,47) 0,158 (1�,3�)MC + hexahloran heptachlor + γ-rays ( 8,25 mSv) 0,14� (23,�8%) 0,147 (25,00) 0,150 (23,47)MC + hexahloran heptachlor + γ-rays ( 17,5 mSv) O,147 (25,00%) 0,12� (34,18) 0,130 (33,67)MC + hexahloran heptachlor + γ-rays ( 35,0 mSv) 0,148 (24,4�%) 0,117 (40,31) 0,121 (38,27)

Joint action of γ – rays and pesticides on some bioenergetic and genetic functions of model...

387Proceedings of the III Congress of Ecologists of Macedonia

This effect is unspecific and most probably is due to the reduce of the adaptive mechanisms of the organ-isms which intensifies the result of the effects of the pesticides introduced in the organisms.

Moreover, the different degree of the syner-getic effect of the γ –rays and the pesticides depends on the rank position and the sex of the individuals in the population which may cause disturbance in the ethological and sex structures and reducing the ge-netic heterogeneity and gene reserves of the popu-lation. Presumably, the different individual change-ability of the organisms entering in the composi-tion of the nature populations, however, at a certain combination of the effect of the anthropogenic fac-tors can prove in some cases a source of elementary evolutional processes causing some changes in the genefund of the population as a whole.

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Fig. 3. ATP-hase activity of the intact liver mitochondria of white mice, treated with linden and γ-rays Сл. 3. ATP-азна активност на интактна митоходрија од слезината кај белиот глушец, третирани со

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