Морфо-функціональні зміни клітин інфузорій Tetrahymena pyriformis W за впливу регуляторів росту рослин — похідних n-оксид піридину

  • Автори: О.П. Васецька
  • УДК: 615.9+661.162.6+547.823+593.17
  • DOI: 10.33273/2663-4570-2021-90-1-40-50
Завантажити прикріплення:

ДП «Науковий центр превентивної токсикології, харчової та хімічної безпеки імені академіка Л.І. Медведя МОЗ України», м. Київ, Україна

 

РЕЗЮМЕ. Мета. Визначити морфологічні зміни інфузорій Tetrahymena pyriformis W за гострого впливу деяких регуляторів росту рослин (РРР) — похідних N-оксид піридину та зіставити їх з функціональними порушеннями клітин.

Матеріали та методи. У роботі використані N-оксид-2-метилпіридин, N-оксид-2,6-диметилпіридин та їх комплекси з органічними кислотами (бурштиновою, малеїновою) або солями металів (ZnCl2, ZnI2, CoCl2, MnCl 2) (всього 15 речовин), синтезованих в Інституті біоорганічної хімії та нафтохімії НАН України. Дослідження проведені на інфузоріях Tetrahymena pyriformis W в ізотоксичних дозах — на рівні токсичних концентрацій — ЛК50, ЛК16 і недіючих концентрацій (ЛК 0). Морфологічні зміни клітин інфузорій оцінювали візуально за допомогою світлового мікроскопу. Порушення структури інфузорій порівнювали з функціональними змінами клітин (руховою активністю та енергетичним станом), отриманих у тому ж експерименті.

Результати та висновки. Показано, що N-оксид-2-метилпіридин, N-оксид-2,6-диметилпіридин та їх комплекси з органічними кислотами (бурштиновою, малеїновою) або солями металів (ZnCl2, ZnI2, CoCl2, MnCl 2) викликають функціональні та морфоструктурні зміни інфузорій, ступінь яких залежить від діючої концентрації. Порушення морфоструктури інфузорій за впливу досліджених РРР характеризуються зміною форми, збільшенням скорочувальної вакуолі, везикуляцією, пошкодженням цілісності цитоплазматичної оболонки, викидами цитоплазми і структурних елементів клітин у живильне середовище.

Комплекси метильних похідних N-оксид піридину з солями металів у досліджених концентраціях знижують швидкість руху і підвищують витрати енергії на рух, викликають зміни поведінкових реакцій і структури клітин більш, ніж N-оксид-2-метилпіридин, N-оксид-2,6-диметилпіридин та їх комплекси з органічними кислотами. Найвираженіші як функціональні, так і морфологічні зміни інфузорій за впливу досліджених РРР відбуваються в концентраціях відповідних ЛК50. У менших концентраціях спостерігались зміни функціональної активності інфузорій. Співставлення отриманих функціональних і морфоструктурних показників стану інфузорій свідчить, що комплекси метильних похідних N-оксид піридину з солями металів чинять токсичну дію на інфузорії більше, ніж комплекси метильних похідних N-оксид піридину з органічними кислотами. Зниження рухової активності та збільшення енерговитрат на одиницю шляху руху, разом з морфологічними змінами структури клітин, є одним із критеріїв токсичності ксенобіотиків для інфузорій та оцінки їхньої життєздатності.

Ключові слова: метильні похідні N-оксид піридину, Tetrahymena pyriformis W, морфо-функціональні зміни.

Introduction. Large-scale use of chemicals, including pesticides and agrochemicals, for various purposes in the national economy is a real threat because of their potential entry into the environment, as they can harm aquatic and terrestrial bioresources, as well as human health. Today in Ukraine a significant proportion of registered plant growth regulators (PGR) are the drugs based on methyl derivatives of pyridine-N-oxide (ivin, poteitin, kapanin, zeastymulin, agrostimulin, betastimulin, triman, etc.) [1], as they contribute to the increase in crops yield, reduction of the amount of pesticide applied without reducing efficiency, production of ecologically clean agricultural products [2–7].

Complexes of 2-methylpyridine-N-oxide, 2,6-dimethylpyridine-N-oxide with organic acids and metal salts synthesized at the Institute of Bioorganic Chemistry and Petrochemistry of the National Academy of Sciences of Ukraine demonstrated high auxin or cytokinin activity on various crops. According to the parameters of acute toxicity for laboratory animals, they are low- or moderately toxic substances. For 2,6- dimethylpyridine-N-oxide (Ivin) and complex of 2-methylpyridine-N-oxide with MnCl2 (Triman) membrane-associated trophic activity, intensification of protein synthesis processes in animals are proven. The ability, in particular of Ivin, to reduce acute toxicity and toxic effects in the body of rats due to exposure to pesticides of different chemical groups was detected [2, 8–10].

It is known that chemicals, which are characterized by high biological activity when released into the environment, have a pronounced toxic effect on aquatic and terrestrial environments. The most dangerous for the environment are salts of heavy metals, surface active substances (SAS), and pesticides. Thus, heavy metals, accumulating in the soil, have a phytotoxic effect on plants, inhibit metabolic functions, limit growth, cause chlorosis, etc. [11]. Salts of heavy metals have a pronounced toxic effect on cyanobacteria Cynechocystis SP atthe level of 1,5 mkg/l-1, zinc and copper oxides are toxic in low concentrations to marine bacteria Vibrio fischeri and freshwater protozoa Tetrahymena Thermophila [12, 13]. With decreasing aquatic temperatures, the toxicity of heavy metals increases significantly [12]. SAS in high concentrations cause lysis of infusoria, and in low — a toxic effect. This is due to the interaction of SAS with lipid bilayer of membranes, which in its turn, leads to changes of osmotic properties and disruption of cell lysis [14]. Oxidative stress caused by chemicals is also among the reasons of structural changes and impairment of infusoria [15].

Toxic effects of heavy metals, some pesticides and various chemical factors on infusoria and other water bodies are mainly reported in the literature as a concentration-effect relationship with the determination of lethal and effective concentrations [9, 16–21] without studies of structural cell disorders being conducted. For some substances, including pesticides, it is noted that along with physiological changes there are changes in the structure of infusoria (loss of their shape, growth of the contractile vacuole, damage to the integrity of the cytoplasmic membrane and organelles). Inhibition of growth and phagocytic activity of infusoria were observed. Important in the mechanism of toxic action of pesticides on infusoria is the imbalance of antioxidant enzymes [22–24].

The effect of PGRs on the state of infusoria is still insufficiently studied. It is shown that methyl derivatives of pyridine-N-oxide under the acute exposure are moderate or low toxic substances for Tetrahymena Pyriformis W infusoria, in high concentrations they cause functional changes (increase or reduce the frequency and speed, total energy consumption for motion), under chronic exposure the curve of concentration-infusoria growth dependence has a mono-, bi- or polymodal form [9, 20, 21]. Along with functional disorders morphological changes in the structure of cells are an important characteristic of toxic effects on infusoria. So the determination of morphostructural changes in infusoria under the acute exposure to pyridine-N-oxide derivatives based PGRs, which are introduced into agricultural practice in Ukraine is an actual task.

The Aim of the Research. To identify morphological changes in Tetrahymena pyriformis W infusoria under the acute exposure to plant growth regulators (PGR) — derivatives of pyridine-N-oxide and compare them to functional disorders of cells.

Materials and Methods. In the study we used the 2-methylpyridine-N-oxide, 2,6- dimethylpyridine-N-oxide and their complexes with organic acids (succinic, maleic) or metal salts (ZnCl2, ZnI2, CoCl2 , MnCl2) (a total of 15 substances), synthesized at the Institute of Bioorganic chemistry and Petrochemistry, NAS, Ukraine, that showed high growth regulating activity and are recommended for agricultural practice in Ukraine as PGR on different crops.

The study is conducted in Tetrahymena pyriformis W infusoria. Tetrahymena pyriformis W infusoria, as a test system in vitro, is widely used in toxicology as an alternative test object to study the toxicity of many pollutants of water bodies, pesticides, heavy metals, extracts of polymeric materials, preservatives and disinfectants, organic and inorganic compounds [23, 25, 26]. As a rule studies on infusoria are conducted under the criterion of “cell death” to determine the parameters of toxicity and predict the danger to humans and the environment. But in addition to the criterion of “cell death” functional and morphological changes are important characteristics of infusoria, and their determination will complement to existing understanding of toxic effects of the impact of chemicals on the cell body.

Studies of morphological changes in Tetrahymena pyriformis W infusoria were performed in isotoxic concentrations: at the level of toxic concentrations — LC50, LC16 and inactive concentrations (LC 0), which were determined earlier [8] and shown in the table. At high concentrations (at the level of LC50) the microscopic examination of the effect of 2- methylpyridine-N-oxide, 2,6-dimethylpyridine-N-oxide and their complexes with organic acids revealed morphostructural changes in infusoria, which are presented in Fig. 1.

 

Table

 Toxicity values of the researched N-pyridine oxide derivatives

 

 

As can be seen in Fig. 1, intact (control) infusoria have a spindle-shaped or slightly elongated body shape with a clearly defined smooth cytoplasmic membrane and well-visible cell organelles. In the upper left part of the figure there are the cells which are in the process of division. The cells moved linearly, the trajectory changed by a sharp turn, spiral swimming was observed.

Under the influence of 2-methylpyridine-N-oxide in high concentrations (at the level of LC50) the body of infusoria in most cases was elongated, some cells had a pear-shaped body. In cells that are located in the centre of the figure the vesiculation of the lateral surface of the body was detected, vesicles are arranged in chain. There was a slowdown in the speed of movement alongside with frequent changes of direction, reduction in the number of food vacuoles. Morphological transformation of body shape of infusoria, which was accompanied by frequent changes of direction, can be the evidence of the change of taxis [27].

Under the influence of the complex of 2-methylpyridine-N-oxide with succinic acid some cells become elongated in the forebody and curved towards the end or pulled out like “proboscis” (the “sickle shaped” form). These cells forebodies seemed to be fixed to glass slides and they quickly moved clockwise around its radial axis. The cytoplasmic membrane was not damaged.

Under the influence of the complex of di-2-methylpyridine-N-oxide with succinic acid, the nature of morphological changes was similar to the complex of 2-methylpyridine-N- oxide with succinic acid, but the field of view was dominated by cells with elongated and sickle shaped body form, which quickly rotated around its axis. Contractile vacuoles were empty, food vacuoles were not traced. The cytoplasmic membrane was not damaged. Atrophied cells were observed.

Under the influence of 2,6-dimethylpyridine-N-oxide in most cells the body shape of infusoria was spindle- or pear-like, some parts of the cell increased in volume, and membrane of some of these cells was deformed, vacuolisation of the cell wall in a small rounded form or in a form of single large spherical mass of ectoplasm, emissions of vesicles into the incubation medium were observed. Food vacuoles were seen clearly.

In infusoria under the exposure to complexes of 2,6-dimethylpyridine-N-oxide with succinic acid and di-2,6-dimethylpyridine-N-oxide with succinic acid the morphostructural changes were similar. In both cases, there were infusoria with distorted body shape (retortor bottle-shaped), but in most of them the structural elements of the cells were well traced. In some cells the integrity damage of the cytoplasmic membrane and the release of cytoplasm into the nutrient medium were observed. Significant morphological changes in infusoria were also observed due to the effect of complexes of 2,6-dimethylpyridine-N-oxide with maleic acid. Some infusoria and contractile vacuole increased in size. Cell vacuolisation, damaged cell membrane, the release of the cytoplasm and cell contents in the nutrient medium were observed.

As can be seen in Fig. 2, under the influence of 2-methylpyridine-N-oxide, 2,6-dimethyl- pyridine-N-oxide in the concentrations corresponding to LC 0 there was a probable increase in the velocity of infusoria; an insignificant increase in this indicator under the influence of complexes of 2-methylpyridine-N-oxide with succinic acid and 2,6-dimethylpyridine-N-oxide with maleic acid was observed. Probable changes in the total energy consumption for the movement of infusoria were not detected, but there was a slight increase in energy consumption per a unit of a path of motion with the greatest effect for 2-methylpyridine-N-oxide and its complex with succinic acid.

At concentrations corresponding to LC16, a probable increase in the velocity of infusoria, the total energy consumption for infusoria movement, and an increase in energy consumption per a unit of a path of motion were observed only for 2-methylpyridine-N-oxide with succinic acid.

In average lethal concentrations under the influence of all studied PGRs a noticeable reduction in speed and increase in energy consumption per a unit of a path of motion were observed. Thus, reduction of speed of movement ranged from 22,4% to 51,4% and was the least expressed under the influence of di-2,6-dimethylpyridine-N-dioxide with succinic acid and most pronounced under the impact of 2,6-dimethylpyridine-N-oxide and its complex with succinic acid. Energy consumption per a unit of a path of motion increased from 47,84% to 127,3%, and the changes were the least expressed under the impact of 2,6- dimethylpyridine-N-oxide with succinic acid and 2,6-dimethylpyridine-N-oxide with maleic acid and the most expressed under the impact of di-2,6-dimethylpyridine-N-oxide with succinic acid, 2-methylpyridine-N-oxide and its complex with succinic acid.

 

Fig. 1. Morphostructural changes in Tetrahymena pyriformis W infusoria under the influence of methyl derivatives of pyridine-N-oxide and their complexes with organic acids at concentrations corresponding to LC50. (1 — a cell at a stage of division, 2 — ectoplasm emission, 3 — vacuolisation)

 

Studies indicate that given methyl derivatives of pyridine-N-oxide and their complexes with organic acids in concentrations at the level of LC 0 and LC16 do not cause morphological changes in the structure of infusoria and cause a slight increase in speed of movement and energy consumption. At average lethal concentrations, they significantly reduce the speed of movement and increase the consumption of energy, cause changes in behavioural reactions and cell structure, which leads to a decrease in the viability of infusoria.

Under the influence of the complexes of 2-methylpyridine-N-oxide and 2,6-dimethyl- pyridine-N-oxide with metal salts, the behaviour of Tetrahymena pyriformis W infusoria in the nutrient medium was the same and depended on the active concentration of substances.

Under the action of low concentrations (at the level of LC0 and LC16) visible morphological changes of infusoria cells were not detected, the movement slowed down, the body shape of infusoria did not differ from the control sample.

As can be seen in Fig. 3, at high concentrations (at LC50) for the complexes of 2,6- dimethylpyridine-N-oxide and 2-methylpyridine-N-oxide with metal salts morphological changes of infusoria cells are more pronounced than for the impact of complexes with organic acids.

Thus, when the impact of the mixture of di-2-methylpyridine-N-oxide with ZnCl2 the body shape of the cells was pear-like, contractile vacuole increased and the some cells were deformed (with elongated and narrowed forebody and extended rear body with an enlarged contractile vacuole, near which other structural elements of the cell were concentrated). The integrity of the cytoplasmic membrane of cells was not damaged.

Under the impact of di-2-methylpyridine-N-oxide complex with ZnI 2 infusoria of different shapes were found: pear-shaped, increased in size with a large contractile vacuole, elongated and flattened laterally, at the bottom there were infusoria with defective cytoplasmic membrane.

The complex of 2-methylpyridine-N-oxide with CoCl2 had a pronounced damaging effect. Most of the cells were deformed, the cytoplasmic membrane of the cells was damaged, and the remains of damaged cells and structural components were observed in the nutrient medium.

Due to the effect of the mixture of di-2-methylpyridine-N-oxide with CoI 2, the shape of most cells was deformed (oval, round, “sickle-shaped”, “cone-shaped“). There were cells with signs of atrophy, in some cells there was a protrusion of the cytoplasm and the integrity of the cytoplasmic membrane was damaged.

Due to exposure to the complex of 2-methylpyridine-N-oxide with MnCl 2 infusoria body shape was mostly elongated, there were cells of “sickle-like” and rounded shapes. Cells with cytoplasmic release into the nutrient medium and with cytoplasmic membrane damage were observed.

The effect on infusoria of the complexes of di-2,6-dimethylpyridine-N-oxide with ZnCl2, as well as of di-2-methylpyridine-N-oxide with CoI 2, was characterized by a significant change in the body shape of most cells. The body shape of the infusoria was deformed (flattened, elongated, cylindrical), the cytoplasmic membrane looked tortuous, and some cells had signs of dehydration. The damage to the integrity of the cytoplasmic membrane of cells, cytoplasmic emissions and cell contents emissions into the nutrient medium were recorded. The contractile vacuole was without any inclusions.

Due to the effect of the complex of di-2,6-dimethylpyridine-N-oxide with ZnI2, infusoria also were of a different shape (pear-shaped, elongated, rounded, horseshoe-shaped). The contractile vacuole was without any inclusions. The vesiculation of cytoplasm, cytoplasm emission in nutrient medium and damage to the integrity of the cytoplasmic membrane of cells were detected.

Complex of di-2,6-dimethylpyridine-N-oxide with CoCl2 in most cells caused form deformation, damage to the integrity of the cytoplasmic membrane, vesiculation of cytoplasm and its release into the nutrient medium; damaged infusoria and the remains of the structural components of cells were observed.

As can be seen in Fig. 4, there was likely reduction in the the speed of movement of infusoria under the impact of all complexes with methyl derivatives of pyridine-N-oxide with metal salts in all concentrations studied, except for the complex of 2-methylpyridine- N-oxide with ZnI 2. The severity of the effect depended on the active concentration of the test substance.

At the lowest concentrations corresponding to LC0, the least expressed changes in the speed of movement of infusoria were identified under the impact of complexes of di-2,6- dimethylpyridine-N-oxide with ZnI 2, 2-methylpyridine-N-oxide with MnCl2, 2-methylpyridine-N-oxide of CoCl2, di-2-methylpyridine-N-oxide with ZnI2 and CoI2, and the most expressed under the influence of complexes of di-2,6-dimethylpyridine-N-oxide with ZnCl 2 and of di-2-methylpyridine-N-oxide with ZnCl2.

 

Fig. 2. Velocity and energy consumption per a unit of a path of motion of Tetrahymena pyriformis W infusoria under the influence of methyl derivatives of pyridine-N-oxide and their complexes with organic acids (* — P ≤ 0.005):

  1. — 2-methylpyridine-N-oxide,
  2. — 2-methylpyridine-N-oxide with succinic acid,
  3. — di-2-methylpyridine-N-oxide with succinic acid,
  4. — 2.6-dimethylpyridine-N-oxide,
  5. — 2.6-dimethylpyridine-N-oxide with succinic acid,
  6. — di-2.6-dimethylpyridine-N-oxide with succinic acid,
  7. — 2.6-dimethylpyridine-N-oxide with maleic acid.

 

Fig. 3. Morphostructural changes in Tetrahymena pyriformis W infusoria under the influence of methyl derivatives of pyridine-N-oxide and their complexes with salts of metals at concentrations corresponding to LC50.(1 — increased vacuole, 2 — elongated body in the shape f proboscis, 3 — damaged membrane (dehydrated cell), 4 — cytoplasm and vacuole emission, 5 — destroyed cells)

 

At concentrations corresponding to LC16, the least pronounced decrease in the velocity of infusoria was due to the effect of complexes of di-2,6-dimethylpyridine-N-oxide with ZnI2, di-2-methylpyridine-N-oxide with ZnCl2 and 2-methylpyridine-N-oxide with CoCl2, and the most pronounced decrease in the speed of movement was under the influence of complexes of di-2,6-dimethylpyridine-N-oxide with ZnCl2, di-2,6-dimethylpyridine-N-oxide with CoCl2.

At these same concentrations increase in movement energy consumption was not expressed, major changes were observed under the impact of complexes of 2-methylpyridine-N-oxide with MnCl2, di-2,6-dimethylpyridine-N-oxide with ZnI2. Under the influence of di-2-methylpyridine-N-oxide complex with ZnI2, unlike other complexes, decrease in movement energy consumption was observed. Under the influence of average lethal concentrations of the studied complexes of methyl derivatives of pyridine-N-oxide with metal salts, the decrease in the speed of infusoria ranged from 47,1% to 77,0% and was most pronounced in di-2,6-dimethylpyridine-N-oxide with ZnCl2, 2-methylpyridine-N-oxide with CoCl2 and di-2,6-dimethylpyridine-N-oxide with CoCl2. The increase in energy consumption per a unit of a path of motion ranged from 40,0% to 90,2% and was most pronounced under the influence of the complexes of di-2-methylpyridine-N-oxide with ZnCl2, di-2,6-dimethylpyridine-N-oxide with ZnCl2, di-2,6-dimethylpyridine-N-oxide with ZnI2, 2-methylpyridine-N-oxide with CoCl2 and 2-methylpyridine-N-oxide with MnCl2, di-2,6-dimethylpyridine-N-oxide with CoCl2. For the complexes of di-2-methylpyridine-N-oxide with ZnI 2 and with CoI2 a decrease in energy consumption per a unit of a path of motion was observed.

 

Fig. 4. Velocity and energy consumption per a unit of a path of motion of Tetrahymena pyriformis W infusoria under the influence of methyl derivatives of pyridine-N-oxide and their complexes with metal salts (* — P ≤ 0.005): 8 — di-2-methylpyridine-N-oxide with ZnCl2, 9 — di-2-methylpyridine-N-oxide with ZnI2, 10 — 2-methylpyridine-N-oxide with CoCl2, 11 — di-2-methylpyridine-N-oxide with CoI2, 12 — 2-methylpyridine-N-oxide with MnCl2,13 — di-2.6-dimethylpyridine-N-oxide with ZnCl2, 14 — di-2.6-dimethylpyridine-N-oxide with ZnI2, 15 — di-2.6-dimethylpyridine-N-oxide with CoCl2

 

As it was discovered in previous studies [20] and is shown in Fig. 2 and Fig. 4, 2- methylpyridine-N-oxide, 2,6-dimethylpyridine-N-oxide and their complexes with organic acids and metal salts are characterized by an increase or decrease in the frequency and speed of movement of infusoria, which reflects the response of cells to a chemical factor. The frequency and speed of infusoria under the impact of the researched methyl derivatives of pyridine-N-oxide and their complexes with organic acids reduced only at concentrations corresponding to LC50, under the influence of complexes of methyl derivatives of pyridine-N-oxide with metal salts — in all studied concentrations with a maximum effect at average lethal concentrations. Differences in energy consumption per a unit of a path of motion are also discovered. Under the influence of 2-methylpyridine-N-oxide, 2,6-dimethylpyridine-N-oxide and their complexes with organic acids an increase in energy consumption for infusoria movement with the greatest effect in average lethal concentrations. Complexes of methyl derivatives of pyridine-N-oxide with metal salts did not increase this indicator as significantly as complexes with organic acids, and under the exposure to complexes of di-2-methylpyridine-N-oxide with ZnI 2 (at 2 higher concentrations) and with CoI2 (at the average lethal concentration) there was a decrease in movement energy consumption.

Compared to the complexes of methyl derivatives of pyridine-N-oxide with organic acids, complexes with metal salts demonstrate a significant reduction in the speed of movement of infusoria and energy consumption for movement which indicates a more toxic effect of the latter.

Impact on morphostructures of infusoria of methyl derivatives of pyridine-N-oxide and their complexes with organic acids and salts of metals is characterized by a change in shape and impaired osmoregulation of cells, increase of contractile vacuole, vesiculation, damage to the integrity of the cytoplasmic membrane, cytoplasm and structural elements of cells emissions into a nutrient medium. However, under the influence of complexes of methyl derivatives of pyridine-N-oxide with salts of metals a change of body shape of infusoria and destructive changes occur more often than in complexes of methyl derivatives of pyridine-N-oxide with organic acids. It should be mentioned that “sickle-like” body shape of infusoria and rapid clockwise movement of these cells was unique to 2-methylpyridine-N-oxide and its complexes with organic acids and salts of metals. For most PGRs studied at average lethal concentrations an increase in the degree of functional changes infusoria was consistent with the expressed structural changes.

Conclusions.

  1. Morphostructural changes in infusoria under the influence of the researched PGRs are characterized by the change in shape, increase of the contractile vacuole, vesiculation, and damage to the integrity of the cytoplasmic membrane, cytoplasm and structural elements of cells emissions into a nutrient medium.
  2. Complexes of methyl derivatives of pyridine-N-oxide with salts of metals in the studied concentrations reduce the speed of movement and increase movement energy expenditure, cause changes in behavioural responses and cell structure to a greater extent than complexes of 2-methylpyridine-N-oxide, 2,6-dimethylpyridine-N-oxide with organic acids.
  3. The most pronounced functional and morphological changes in infusoria under the influence of the studied PGRs occurred at concentrations corresponding to LC50. Changes in the functional activity of infusoria were observed at lower concentrations.
  4. Comparison of the obtained functional and morphostructural indicators of infusoria shows that complexes of methyl derivatives of pyridine-N-oxide with metal salts have greater toxic effects on infusoria than the complexes of methyl derivatives of pyridine-N-oxide with organic acids.
  5. Decrease in motor activity and increase in energy consumption per a unit of a path of motion together with morphological changes of cell structure are the criteria of xenobiotic toxicity for infusoria and assessment of their viability.

 

REFERENCES

  1. Perelik pestytsydiv i ahrokhimikativ, dozvolenykh do vykorystannya v Ukrayini. [Addendum to the List of pesticides and agrochemicals authorised for use in Ukraine: Special edition of journal “Propozytsiia”]. Spetsialʹnyy vypusk zhurnalu „Propozytsiya”; K.: Yunivest Media; 2019, 1039 p.

  2. New plant growth regulators: basic research and technologyes of application. Monograph. Ponomarenko SP, Iutynska HO, editors. Kyiv: Nichlava; 2011, 210 р.

  3. Ponomarenko SP, Tsyhankova VA, Blyum YAB, Halkin AP. Novyy napryamok u roslynnytstvi — zastosuvannya pryrodnykh polikomponentnykh rehulyatoriv rostu roslyn z biozakhysnym efektom [Recent trend in plant growing — use of natural multicomponent plant growth regulators with bioprotective effect]. Nauka ta innovatsiyi. 2013; 9(5): 69–77.

  4. Tsyhankova VA, Andrusevych YAV, Babayants OV, Ponomarenko SP, Medkov AI, Halkin AP. Pidvyshchennya rehulyatoramy rostu imunitetu roslyn do patohennykh hrybiv, shkidnykiv i nematod [Increase in plant immunity to pathogenic fungi, vermin, nematodes by growth regulators]. Fiziolohiya ta biokhimiya kulʹturnykh Roslyn. 2013; 45(2): 138–47.

  5. Babayants OV, Tsyhankova VA, Ponomarenko SP, Medkov AY. Rolʹ rehulyatorov rosta v immuno-zashchytnykh reaktsyyakh rastenyy na bolezny, vyzvannykh patohennymy orhanyzmamy [Role of growth regulators in plant immunity protection against diseases induced by pathogenic organisms]. Posibnyk ukrayinsʹkoho khliboroba. 2014; 1; 161–5.

  6. Tytov VN, Smyslov DH, Dmytryeva HA, Bolotova VY. Rehulyatory rosta rastenyy kak byolohycheskyy faktor snyzhenyya urovnya tyazhelykh metallov v rastenyy [Plant growth regulators as biological factor in the reduction of the level of heavy metals in plants]. Vestnyk OrelHAU. 2011; 4(31): 4–7.

  7. Ponomarenko SP. Biostymulyatory rostu. Shlyakh do ekolohichno chystoyi syrovyny dlya vyhotovlennya produktiv dytyachoho kharchuvannya [Growth biostimulators. Strategies of environmentally safe raw materials for production of baby food products]. Zakhyst Roslyn. 1998; 4: 21.

  8. Vasetska OP. Hostra toksychnistʹ novykh rehulyatoriv rostu roslyn — pokhidnykh N-oksyd pirydynu [Acute toxicity of the new plant growth regulators — derivatives of pyridine N-oxide]. «Suchasni problemy toksykolohiyi, kharchovoyi ta khimichnoyi bezpeky». 2016; 3(75): 5–11.

  9. Vasetska OP, Zhminko HG. «Paradoksalʹnye» éffekty v toksykolohyy, mekhanyzmy y metodycheskye podkhody k ykh prohnozyrovanyyu (po dannym lyteratury y sobstvennykh yssledovanyy) ["Paradoxical" effects in toxicology, mechanisms and methodological approaches to prediction (according to the literature and our own research)]. «Suchasni problemy toksykolohiyi, kharchovoyi ta khimichnoyi bezpeky». 2015; 1/2 (68/69): 54–66.

  10. Vasetska OP. Kombinovana diya rehulyatoriv rostu roslyn na osnovi pokhidnykh N-oksyd pirydynu ta deyakykh pestytsydiv riznykh khimichnykh hrup [Combined effect of plant growth regulators based on pyridine n-oxide derivatives and some pesticides of different chemical groups]. Suchasni problemy toksykolohiyi, kharchovoyi ta khimichnoyi bezpeky. 2017; 3: 26–33.

  11. Yadav SK. Heavy Metals Toxicity in Plants: An Overview on the Role of Glutathione and Phytochelatins in Heavy Metal Stress Tolerance of Plants. South African Journal of Botany. 2010; 76: 167–79. DOI: 10.1016/j.sajb.2009.10.007.

  12. Bohacheva AS, Shylov VV, Polozova YeV. Chuvstvytelʹnostʹ tsyanobakteryy Cynechocystis SP k toksycheskomu deystvyyu soley tyazhelykh metallov [Sensitivity of cyanobacteria Cynechocystis SP to the toxic action of heavy metal salts. Current issues of toxicology and radiobiology. Russian Scientific Conference with International Participation]. Aktualʹnye problemy toksykolohyy y radyobyolohyy. Rossyyskaya nauchnaya konferentsyya s mezhdunarodnym uchastyem; 2011 maya 19–20; Sankt-Peterburh. Sankt-Peterburh: “Folyant”; s. 26.

  13. Kurvet I, Juganson K, Vija H, Sihtmäe M, Blinova I, Syvertsen-Wiig G, et al. Toxicity of Nine (Doped) Rare Earth Metal Oxides and Respective Individual Metals to Aquatic Microorganisms Vibrio fischeri and Tetrahymena thermophila. Materials. 2017; 10(7), 754. https://doi.org/10.3390/ma10070754.

  14. Prisnyi AV, Volynkin YUL, Kampos NN. Mekhanizmy ustoychivosti infuzoriy k khimicheskim povrezhdeniyam i ikh preodoleniye letal'nymi kontsentratsiyami sinteticheskikh poverkhnostno aktivnykh veshchestv (SPAV) [The resistance’s mechanisms of infusorians to chemical damages and their overcoming by lethal concentration of synthetic superficially active substances (SSAS)]. Nauchnyye Vedomosti, 2009; 11(66): 45–54.

  15. Mar PD, Khalfi BE, Soukr A. Protective effect of oregano and sage essentials oils against the effect of extracellular H2O2 and SNP in Tetrahymena thermophila and Tetrahymena pyriformis. Journal of King Saud University — Science. 2020; 32(1): 279–87. https://doi.org/ 10.1016/ j.jksus.2018.05.005.

  16. Garad U, Desai SN, Desai PV. Toxic effects of monocrotophos on Paramecium caudatum. African J. Of Biotechnology. 2007; 6(19): 2245–50.

  17. Amara A, Quiniou F, Durand G, Bour ME, Boudabous A, Hourmant A. Toxicity of Epoxiconazole to the Marine Diatom Chaetoceros calcitrans: Influence of Growth Conditions and Algal Development Stage. Water Air Soil Pollution. 2013; 224, 1417. https://doi.org/10.1007/s11270-012-1417-9.

  18. Cheremnykh ЕG, Kuleshin АV, Kuleshina ON. Biotestirovaniye pishchevykh dobavok na infuzoriyakh [Screening of foods additives on infusoria]. Vestnik RUDN, seria Ekologiya i bezopasnost' zhiznedeyatel'nosti. 2011; 3: 5–12.

  19. Fedotov AS. Otsenka toksichnosti ingibitorov korrozii Azol Cl-130, Azol-5010 marki A i Azol-5010 marki V dlya gidrobiontov [Toxicity assessment of corrosion inhibitors Azol CI-130, Azol 5010, brand A and Azol 5010, brand B for hydrobionts]. Toksikologicheskiy vestnik. 2013; 5: 38–44.

  20. Vasetska ОP. Vplyv novykh rehulyatoriv rostu Roslyn — pokhidnykh n-oksyd pirydynu na rukhovu aktyvnistʹ i enerhetychnyy stan infuzoriy Tetrahymena pyriformis W. [Effect of the new plant growth regulators — N-oxide pyridine derivatives on mobile activity and energy state of infusoria Tetrahymena pyriformis W]. Suchasni problemy toksykolohiyi, kharchovoyi ta khimichnoyi bezpeky. 2018; 2–3: 42–50.

  21. Zhmínko OP, Prodanchuk MG. Vpliv deyakikh pokhídnikh N-oksid píridinu na ríst populyatsíí̈ ínfuzoríy Tetrahymena pyriformis W. [Influence of derivative N-oxide pyridine on growth of infusorians population Tetrahymena pyriformis W]. Sovremennyye problemy toksikologii. 2002; 2: 33–7.

  22. Maurya R, Dubey K, Singh D, Jain AK, Pandey AK. Effect of difenoconazole fungicide on physiological responses and ultrastructural modifications in model organism Tetrahymena pyriformis. Ecotoxicology and Enviro-nmental Safety. 2019; 182: 109375. https://doi.org/10.1016/j.ecoenv.2019.109375

  23. Maurya R, Pandey AK. Importance of protozoa Tetrahymena in toxicological studies: A review. Science of The Total Environment. 2020; 741: 140058. https://doi.org/ 10.1016/j.scitotenv.2020.140058

  24. Moumeni O, Berrebbah H, Azzouz Z, Amamra R, Otmani H, Alayat A, et al. Effects of Cycloxydim on Population Growth, Phagocytosis, Contractile Vacuole Activity and Antioxidant Responses in the Freshwater Ciliate (Paramecium tetraurelia). Research Journal of Environmental Toxicology. 2016; 10: 115–25. DOI: 10.3923/rjet.2016.115.125

  25. Kompleksnaya biologicheskaya otsenka ob"yektov prirodnogo i iskusstvennogo proiskhozhdeniya na Tetrahymena pyriformis W. Metodicheskiye rekomendatsii; Minsk; 1996, 19 s.

  26. Zhmínko PG, Yankevich MV, Gerasimova VG, Lisenko KO, Zhmínko OP. “Metodika yekspresnogo bíotestuvannya khímíchnikh rechovin, sintetichnikh materíalív gospodarskogo, pobutovogo ta medichnogo priznachennya y otsínki íkh nebezpeki dlya lyudini”. Reestr galuzevikh novovveden. 14–15, 45/14/01.

  27. Bubnov AG, Buynova SA, Gushchin AA, Izvekova TV. Biotestovyy analiz — integral'nyy metod otsenki kachestva ob"yektov okruzhayushchey sredy: Uchebnoye posobiye. [Biotest analysis — an integral method for assessing the quality of environmental objects: Textbook]. Ivanovo: Izd-vo GOU VPO Ivan. gos. khim-tekhnol. un-t; 2007, 112.

 

Received 10/26/2020