Method to assess toxic effect of pesticides at water objects

FIELD: ecology.

SUBSTANCE: during realisation of the method the test objects are soaked in tested solutions, parameters of test objects survivability are registered, and using them, threshold concentrations of tested pesticide toxicity are calculated, besides, pathomorphological modifications are registered in test objects, the average percentage of malformations is calculated, and the threshold concentration of teratogenic impact is established as the pesticide concentration with minimum teratogenic impact at test objects, and extent of pesticides toxicity is estimated on the basis of the threshold concentration coefficient, which is calculated using the formula Knemb - coefficient of threshold concentrations of pesticides toxicity taking into account their teratogenicity, LC16 - limit concentration of pesticides toxicity causing death of 16% embryos, EterC16 - threshold concentration of teratogenic effect of pesticides. At the same time if values of the coefficient Knemb >10, the class of pesticides hazard is established as I, i.e. extremely hazardous, 5-10 - the class of pesticides hazard is set as II, i.e. highly hazardous, 1-5 - the class of pesticides hazard is established as III, i.e. hazardous, <1 - the class of pesticides hazard is IV, i.e. moderately hazardous. Test objects are embryos of sturgeons.

EFFECT: increased accuracy and validity of assessment.

2 cl, 1 ex, 6 tbl

 

The present invention relates to research in the field of environmental protection, aquatic toxicology, and in particular to methods Toxicological evaluation of the action of pesticides on water bodies that have significance for fisheries. The method can be used for tightening of the class of ecological risks of pesticides in the regulation of their content in water and fisheries.

The main way to prevent water pollution by pesticides is a preliminary determination of the degree of toxicity to aquatic organisms of each of the promising pesticides and the prohibition of the use within the protection of the fisheries of those areas, which are characterized by high toxicity to aquatic organisms.

This requires the development of fast, cheap and effective methods of preliminary determination of toxicity of pesticides.

There is a method of determining the toxicity of chemicals (A.S. USSR №1564539, IPC 5 G01N 33/18, 33/00) [1], including the cultivation of the strain Saccharomyces cerevisial 15-A4 on liquid nutrient medium, the effect of the analyte and the subsequent evaluation of the results.

There is a method of quantitative determination of organophosphorus pesticides (A.S. USSR №1111270, IPC 4 A012K 61/00, G01N 33/18) [2], providing space in the analyzed solution as a TEC the objects Daphnia (Daphnia magna), accounting for the length of their survival at 34-36°C and then determining the amount of pesticide on the calibration curve.

There is a method of determining the toxicity of aquatic environments (A.S. USSR №1270699, IPC 4 G01N 33/18, C02F 3/32) [3], including the study of adaptation in animals gastropods, pre-calibrated degree of sensitivity and resistance of aquatic organisms to a reference toxicant, test impacts experienced a series of aquatic organisms to specific concentrations of the studied environments, ensuring the constancy of external conditions and stepwise increasing concentrations when exposed to each of the following series of hydrobionts, calculating her motor activity for each concentration and the extent of toxicity aquatic environments.

Existing classification to establish the hazard class of pesticides for aquatic ecosystems is based on toxicity data on the value of the MPC, the stability of the substance in the reservoir and its cumulative properties (Methodical recommendations on the establishment of ecological fishery regulations (MAC and Asil) of pollutants for water with fishery value. - M.: VNIRO, 1998. - 147 C.) [4].

At the same time, according to the "Methodological recommendations" [4], approved by the Ministry of agricultureeconomy and food of the Russian Federation in 1998, when ecology-fisheries regulation of pesticides is required experiments to identify specific effects - mutagenicity, cytogenotoxicity, teratogenicity of chemicals to aquatic organisms. Known methods of assessment of specific effects (Mutagenic substances in fishery water bodies. Methodological guidance on the assessment of genotoxicity and cytotoxicity in the development of the fisheries regulations (MPC). 1991) [5], (Detlef T.A., A.S. Ginsburg, I. Schmalhausen Development of sturgeons. - M.: Nauka, 1981) [6].

The results of many years of research have shown that the presence of mutagenic properties and genotoxicity in pesticide recent generations is the exception. However, recorded that the majority of modern pesticides new generations in certain doses have teratogenic activity for fish in the early stages of development (embryos and prelicense). At the same time when setting class environmental hazards of pesticides to aquatic ecosystems, the degree teratogenic hazard to aquatic organisms is not considered, that is a disadvantage of the known classification.

It is established that the most vulnerable to pollution and other changes in environmental conditions are the animals on the juvenile stage of development. These include eggs and fish larvae. It is known that eggs and larvae due to the lack of development of protection systems and the impossibility of leaving the contaminated zones can be most susceptible to pesticide intoxication.

When assessing pesticide toxicity to fish in the early stages of ontogenesis, it is important to have a complete picture of the disorders that causes a particular pesticide.

The closest to the invention is selected as a prototype, a method of evaluating the toxicity of pollutants to waters of the far Eastern seas (patent RU №2215290, IPC 7 G01N 33/18) [7], in which the test objects, which are used prelicense Japanese anchovy, soak in the test solution, record the physiological response and the degree of toxicity of the pollutant judged on Toxicological parameters - LC0LC16LC50LC84LC100where LC16considered to be the threshold concentration (upper level concentration that causes a loss of 16% of embryos).

In the known method does not take into account the degree of teratogenic risk of toxicants to aquatic organisms, which reduces the reliability of the assessment of toxic effects of pollutants on water objects.

The purpose of the present invention is to improve the accuracy of estimating the toxicity of pesticides to aquatic ecosystems with regard to their teratogenic effect on the embryos sturgeon.

The technical result of the invention is more fully assess the toxicity of pesticides to aquatic ecosystems.

This is achieved by the fact that in the known method OC the NCI toxicity of pesticides to water bodies, in which the test object is kept in the test solution, record the survival rates of the test object, on the basis of which the calculated threshold concentration toxicity testing of pesticides, and assess the degree of toxicity of pesticides, according to the invention additionally register pathological changes in the test object, calculate the average percentage of deformities, and as the threshold concentration teratogenic effects set the concentration of the pesticide, which has less teratogenic effect on the test objects, and on the degree of toxicity of pesticides is judged on the basis of the coefficient of threshold concentration, which is calculated by the formula

where Kpemb- coefficient of threshold concentrations the toxicity of pesticides, taking into account their teratogenicity.

LC16- threshold concentration of the toxicity of pesticides, causing the death of 16% of the embryos.

ETerTo16threshold concentration teratogenic effects of pesticides.

When the values of Kpemb

>10 establish the hazard class of pesticides I, i.e. extremely dangerous.

5-10 installing the hazard class of pesticides II, i.e. high-risk;

1-5 installing the hazard class of pesticides III, i.e. dangerous;

<1 CL the SS dangers of pesticides IV, i.e. moderately dangerous.

As test objects using embryos sturgeon.

Comparison of the prototype and the proposed method shows that the latter differs from the prototype additional registration, accounting and analysis of teratogenic effects of toxicants on the test objects. And the resulting research independent indicators thresholds lethal and teratogenic effects are used together and are used to derive the coefficient of threshold concentrations (pemb). In the calculation of the coefficient (Kpemb) achieving the goal of improving the accuracy and reliability of the assessment of the toxicity of pesticides to fish embryos. Next, on the basis of Kpembclassification of pesticides according to the degree of teratogenic activity of fish during early ontogenesis.

The method is as follows.

Experimental investigations were carried out actions more than 50 pesticides on aquatic organisms. As test objects used sturgeon during early ontogeny - caviar Bester (Huso huso L. X Acipenser ruthenus L.), sturgeon (Acipenser gueldenstaedti), stellate sturgeon (Acipenser stellatus).

In the chemical 2-liter capacity made of the studied pesticides, creating a set of given concentrations. And left control capacity without toxicants. Oplog is solved eggs were placed in Petri dishes and placed in containers with pesticides.

To assess the lethal action of the pesticide recorded survival rates of embryos during embryogenesis. Were conducted systematic observations and records of live and dead embryos for experience, ustanovlena dependence of the survival of organisms depends on the concentration of pesticide and time of its impact. At the end of the experiment was conducted in the calculation of basic toxicometric parameters (CF0LC16LC50LC84LC100using the method of probit analysis Webpronews.com (Prozorovsky V.B. have been Using the method of least squares for probit analysis of the mortality curves. // Pharmacol. and toxicol. - 1962. No. 1) [8]. The estimated value LC16(upper level concentration that causes a loss of 16% of embryos) is considered to be the threshold. The threshold lethal action LC16for all investigated pesticides listed in table 1.

Teratogenic analysis was performed on the stages of development of fish embryos and hatching. Integral index of the teratogenic effects of pesticides served pathological changes in experimental organisms were fixed abnormal development of fertilized eggs and pathomorphological signs of hatched embryos. On the basis of teratogenic analysis was calculated the average percentage of deformities, and as the threshold concentration was at tanawin experimental concentration of the pesticide, providing the minimum teratogenic effect.

Experimental studies have shown that it is not always possible to experimentally determine the threshold concentration teratogenic effects of pesticides. In some cases, the experimentally determined minimum effective concentration by abnormal development of embryos, such as pesticides belonging to the chemical classes of pyrethroids, dithiocarbamates, derivatives of imidazole, benzimidazole and other exceeded 16% of the levels adopted as a threshold in Toxicological experiments (table 1).

In this regard, there is a need to unify the definition of the threshold concentration and obtaining the calculated values to assess the specificity of action of the studied pesticides.

The estimated value of the threshold concentration teratogenic effects of pesticides (eTerTo16), not previously used in embryotoxicity studies, was obtained as a result of mathematical processing of the data by abnormal development of embryos of fish by the method of probit analysis Webpronews.com (8)based on the linear relationship between concentration and effect, expressed in probita. The value of eTerTo16allowed more accurately determine the threshold teratogenic effects of pesticides of different chemical classes is impressive. The values of eTerK16for all experimental pesticides are given in table 1.

Based on the calculated threshold value LC16and eTerTo16was developed formula representing the amount teratogenic effects of pesticides to fish embryos. Measure teratogenicity (specificity steps) of pesticides for fish embryos was determined by the magnitude of the coefficient threshold concentrations (pemb), i.e. ratio of the threshold lethal steps to the threshold of teratogenic effects:

The calculated values Forpemball experimental pesticides are given in table 1.

The value of Kpemballowed to determine the degree of potential danger of certain pesticide chemical class in relation to development opportunities teratogenic effect. The value of Kpembgreater than 1 indicates the electoral effects of pesticides on embryonic development of fish.

The analysis of the presented data in table 1 and the calculated values Topembfor sturgeon embryos allowed us to conclude that the degree of amplification of the teratogenic activity of embryos studied chemical classes of pesticides were located in the following range: derivatives of pyridine < strobilurin < derivatives of the sulfonylureas < produced by the water aryloksyfenoksypropionowe acid < derivatives of N-arylcarbamoyl acid < neonicotinoids, derivatives of imidazole and benzimidazole < triazoles < derivatives of pyrimidine, thiophosphorous and dithiocarbamates acid < pyrethroids.

When the coefficient of threshold concentrations (pemb) was developed classification of pesticides according to the degree of teratogenic risk, presented in table 2. According to the proposed classification of pesticides that have the value of Kpembless than 1 are moderately dangerous for the embryos of fish, if Kpemb1-5 - dangerous,pemb5-10 - high. The value of Kpembexceeding 10 indicates extreme danger of the pesticide.

The technical result of the invention is more fully assess the toxicity of pesticides.

The invention is illustrated by example.

Studies to determine the degree of toxicity of the fungicide Tyr, 425 g/l, TPS at spawn Bester (Huso huso L. X Acipenser ruthenus L.) was performed with concentrations in the range 0.05-5.0 mg/L.

In the experience took artificially fertilized eggs at the stage of 4 blastomeres. Laid out in Petri dishes for 30 pieces in each and were placed in 2-liter containers with solutions fungicide Tyr, which was filed oxygen. The height of the liquid column values were not as favorable conditions ha is obmana germ ensured a continuous supply of oxygen. Water temperature averaged 18.6°C.

The degree of toxicity of different concentrations of the fungicide on developing eggs of Bester was evaluated on the following parameters: survival, duration of the incubation period, the rate of passage stages, pathomorphological signs of embryos and hatched non-feeding larvae (teratology analysis). In the experiment conducted daily accounting of the dead embryos were fixed stage of development.

On the basis of the data on survival of fertilized eggs Baster in solutions of 0.05-5.0 mg/l calculated toxicometric parameters presented in table 3. Calculation toxicometric parameters for pesticide embryos Baster produced with less natural death in control and experience (amendment Abbott).

Data on dynamics of hatching and duration of the incubation period, the embryos Baster presented in table 4.

In solutions with a maximum concentration of fungicide Tyr 5.0 mg/l the development of embryos continued until stage 15 (stage middle gastrula), then came their death.

During observations of developing embryos Baster in fungicide solutions in the concentration range 0.1-2.5 mg/l revealed teratogenic effect.

The pesticide at concentrations of 0.5-2.5 mg/l had a strong toxic effect, causing various disorders embryogenesis, C is the delay hatching, the lengthiness of it in time. In solutions with the above concentrations of the fungicide in embryos was observed violations of the process of gastrulation, which was manifested in the delay fouling dark vegetative cells and preservation of the yolk tube of considerable size. Later this led to bookmark shortened, twisted nerve records. Along with this, at the stages of organogenesis specific violations typical of pesticides derived dithiocarbamates acid. Such disorders include severe flooding renal collecting ducts, the accumulation of fluid in the cavity of the medulla oblongata, pericardial cavity, a kind of multiple curvature of the chord, which gives it a "corrugated". In solutions with concentrations of 0.5-2.5 mg/l hatched embryos had abnormalities in the structure is shortened, With shaped, curved trunk, "corrugated" chord, cropped tail, dropsy of the pericardial cavity, the underdevelopment of the brain Department.

In solutions with a concentration of fungicide Tyr 0.1 mg/l was also observed above violations embryo development, but the death of the embryos did not exceed natural. Marked delay hatching on the day and the asynchrony of hatching.

In solutions with a concentration of pesticide 0.05 mg/l deviations in the development of embryos from the control group n is observed. The rate of passage stages, single and mass hatching took place synchronously with the control. The hatched embryos pathology was not found.

On the basis teratogenic analysis was estimated percentage of malformations in the embryos Baster before hatching (table 5).

As can be seen from table 5, the threshold concentration of teratogenesis when the concentration of the pesticide 0.05 mg/l for embryos Baster as a result of experimental work was not revealed. At a concentration of 0.1 mg/l (following inactive concentration of 0.05 mg/l) ugly individuals amounted to 50.9%.

Therefore, as the threshold concentration teratogenic effects of fungicide shooting for embryos of Bester was adopted by the estimated value of eTerTo16=0.064 mg/l obtained by mathematical processing of the data by abnormal development of embryos (percentage of deformities) using probit analysis Webpronews.com [Prozorovsky V.B. have been 1962].

Thus, for embryos Baster threshold concentration of fungicide thier survival rates (LX16) amounted to 0.39 mg/l, the threshold concentration for teratogenic effects (eTerTo16) - 0.064 mg/l the Data are shown in table 1.

Next, we calculated the coefficient of threshold concentrations (pemb), which represents the ratio of the threshold lethal action to the threshold teratogen the th steps:

Topemb=0.39:0.06=6.09.

Therefore, according to the proposed classification (table 2) according to the degree of teratogenic risk fungicide Tyr applies to class II (high-risk pesticides).

It should be noted that the hazard class of pesticide is established: a) to determine the extent of environmental hazards of pesticide in connection with its emergence in aquatic ecosystems; b) to establish priority in the control of pollution of the water environment) to justify recommendations for the use of the pesticide. If the pesticide belongs to the 1st class of hazard (extremely hazardous substances of anthropogenic origin), its use within the protection of the fisheries zone is prohibited (Methodical recommendations on the establishment of ecological fishery regulations (MAC and Asil) of pollutants for water with fishery value. - M.: VNIRO, 1998. - 147 C.) [4].

By calculating the value Kpembit was found that the class teratogenic risk of some pesticides (Alto super, Start, Isidor, TMTD, Tyr) was exceeded in accordance with generally accepted classification of hazard of pollutants to aquatic organisms, taking into account the value of the MPC, the accumulation factors (Kn) and stability (τ95). The data presented in t the blitz 6.

Thus, the proposed classification of pesticides according to the degree of teratogenic activity of fish during early ontogeny can be used for tightening hazard class in the regulation of pesticides.

32.94
Table 1
The parameters of the toxicity and teratogenicity of pesticides for embryos sturgeon
Name of pesticide (name of active ingredient)LC16mg/lThe minimum effective concentration, mg/l (% ugly animals)ETerTo16mg/lThe coefficient of threshold concentrations Topemb(hazard class)
12345
Derivatives cyclopropanecarbonyl acid (pyrethroids)
Alfaz (α-cypermetrin)8.690.1 (20%)0.079109.96 (I)
Karachar (λ-cigalotrin) 9.830.5 (at 26.32%)0.4223.40 (I)
Cinfo (β-cyfluthrin+timeout)13.070.5 (14.54)0.5922.15 (I)
Chinook (β-titluri+Imidacloprid)28.451.0 (32.73)0.62At 45.89 (I)
Decis-pros (deltamethrin)0.920.5 (at 22.81%)0.253.68 (III)
Operat (λ-cigalotrin)0.490.5 (23.53%)0.421.17 (III)
Modesto (β-cyfluthrin+clothianidin)7.875.0 (18.87%)4.791.81 (III)
Poncho Beta (β-cyfluthrin+clothianidin)3.811.0 (22.6%)0.774.95 (III)
Talstar (bifenthrin)0.0078 0.01 (33.3%)0.00411.90 (III)
Derivatives tiofosfornoy acid
Finition (fenitrothion)0.840.5 (11.11%)0.541.56 (III)
Fyfanon (Malathion)6.431.0 (17.65%)1.643.92 (III)
Derivatives of phosphorous acid
Efatal (fosetyl aluminum)41.9350.0 (15.22%)52.370.8 (IV)
Derivatives dithiocarbamates acid
TMTD (thiram)0.820.1 (18.9%)0.07910.38 (I)
Tyr (thiram+tebuconazole)0.390.1 (50.9%)0.0646.09 (II)
The polishers (metiram)0.480.25 (7.41) 0.172.82 (III)
Start (thiram+tebuconazole)0.390.1 (28.92%)0.075.57 (II)
Derivatives of N-arylcarbamoyl acids
Befor Expert (desmedipham+phenmedipham+ethofumesate)17.9710.0 (15.09%)12.881.43 (III)
Betharan Extra (desmedipham+phenmedipham+ethofumesate)11.2910.0 (24.4%)8.341.35 (III)
The beater Trio (desmedipham+phenmedipham+ethofumesate)2.412.5 (at 15.97%)2.520.96 (IV)
Leader (desmedipham+phenmedipham+ethofumesate)95.7625.0 (18.87%)23.374.10 (III)
Betanal Quad (desmedipham+phenmedipham+ethofumesate+metamitron)At 23.4330.0 (10.26%)0.71 (IV)
Posvodnie aryloksyfenoksypropionowe acids
AUG-99 (fenoxaprop-p-ethyl)0.761.0 (23.8%)0.770.99 (IV)
Furore-ultra (fenoxaprop-p-ethyl)19.5010.0 (11.76%)10.211.92 (III)
Normal (hisamoto-p-ethyl)0.861.0 (16.98%)0.970.89 (IV)
Derivatives of pyrimidine
Lenacil (lentil)85.4050.0 (32.29%)34.172.50 (III)

Continuation of table 1
12345
Derivatives of pyridine
Dichter With the pen (Diquat) 24.2225.0 (8.89%)45.010.54 (IV)
Golden Ring (Diquat)25.2425.0 (13.33%)31.80.79 (IV)
Diquat (Diquat)20.3750.0 (25%)30.440.67 (IV)
Sukhovey (Diquat)17.4850.0 (11.54%)31.270.56 (IV)
The neonicotinoids (derivatives of imidazole or thiazole)
AC-126 (Imidacloprid)39.8610.0 (34.55%)15.542.56 (III)
Sparkle gold (Imidacloprid)51.3950.0 (23.4%)At 43.961.17 (III)
Iminor (Imidacloprid)65.1310.0 (22.6%)6.3110.32 (I)
Condor (Imidacloprid)45.3750.0 (29.55%)21.832.08 (III)
Of aktar (thiamethoxam)6.405.0 (24.65%)2.581.94 (III)
Apache (clothianidin)8.115.0 (9.26%)6.481.25 (III)
Derivatives of imidazole and benzimidazole
Tapir (imazethapyr)515.59250.0 (5.26%)448.171.15 (III)
Fabian (imazethapyr+chlorimuron-ethyl)528.50100.0 (11.1%)167.763.15 (III)
Vincit Forte (imazalil+thiabendazole+flutriafol)6.6210.0 (8.33%)8.820.75 (IV)
□ trade (imazalil+thiabendazole+flutriafol)14.285.0 (28.3%) 4.043.53 (III)
Derivatives of triazoles
Strike (flutriafol)7.9910.0 (28.6%)6.771.18 (111)
Alcor (tsyprokonazolu)2.851.0 (9.09%)1.771.61 (III)
Alto super (tsyprokonazolu+propiconazole)34.151.0 (10.91%)2.5513.39 (I)
Karamba (metconazole)1.140.5 (11.11%)0.631.81 (III)
Prosaro (prothioconazole+tebuconazole)15.7130.0 (9.68%)At 32.750.48 (IV)
Tabutin (tebuconazole)52.44100.0 (23.33%)80.710.65 (IV)
Derivatives of the sulfonylureas
Singer (metsulfuron-methyl)75.75100.0(15%)186.560.41 (IV)
AUG-65 (tribenuron-methyl)510.8500.0 (13.04%)563.310.91 (IV)
Segment (azimsulfuron-methyl)672.27750.0 (23.08%)662.311.02 (111)
The artenay (metsulfuron-methyl)141.5450.0 (10.91%)75.471.88 (III)
The Saracens (metsulfuron-methyl)122.43100.0 (8.51%)172.60.71 (IV)
Peak (prosulfuron-methyl)>1500750.0 (8.9%)821.91.83 (III)
Pulled (chlorsulfuron-methyl+dicamba)1125.17400.0 (14.04%)415.262.71 (III)
Venison (chlorsulfuron-methyl+dicamba)375.0250.0 (5.45%)365.191.03 (III)
Musket (iodosulfuron-methyl+mefenpyr-diethyl)1.9610.0 (27.27%)7.510.26 (IV)
Strobilurin
But (Trifloxystrobin)0.00170.0025 (14.28%)0.00280.61 (IV)
Cabrito (pyraclostrobin+metiram)0.710.5 (7.41%)0.790.9 (IV)
"Pictor " (dimoxystrobin+boscalid)0.0135.0 (15.56%)5.580.02 (IV)

Table 2
Classification of pesticides according to the degree of teratogenic risk
The coefficient of threshold concentrations Topemb Hazard class
>10I extremely dangerous
5-10II - high
1-5III - hazardous
<1IV - moderately hazardous

Table 3
Survival of embryos Baster in solutions fungicide Tyr
Concentration, mg/lDeath* early instancesDeath adjusted Abbot, %Toxicometric parameters, mg/l
article 5-12 (crushing)article 13-18 (gastrolyte)article 19-29 (neurulation)v.30-35 (before hatching)Only
Control322070LC0/sub> =0.10
LC16=0.39
LC50=1.74
LC84=3.67
LC100=4.74
0.05231170
0.1331070
0.547701820.75
1.0610612330.19
2.56152034469.81
5.01248--60100
Note: - * - the total loss in the three replications of the experiment

Table 4
Dynamics of hatching and the duration of the incubation period, the embryos of Bester during curing in solutions fungicide Tyr
Concentration, mg/lHatching, %The duration of the incubation period, h
singlemass
Control9.43100104
0.0511.32100104
0.103.77132
0.507.14136
1.006.25136

Table 5
Anomalies of the Baster in the embryonic period of development during curing in solutions fungicide Tyr
Concentration, mg/lThe total number of instancesThe number of malformed individuals instancesThe number of malformed individuals %
Embryos, stage 35
Counter.5300
0.055300
0.1532750.9
0.54242100
1.03737100
2.51616100

Table 6
Hazard classes, some pesticides that are installed on the hazard classifications of pollutants and teratogenesis
Name of pesticideMPC, mg/lHazard class
on MAC,nτ95on teratogenesis
Alto Super 330 g/l EC0.00033I
Start 400 g/l, KS0.00023II
Isidor 200 g/l, WRC2.53I
TMTD 400 g/l, TPS0.000253I
Tyr 425 g/l, TPS0.000253II

1. The method of evaluating the toxic effects of pesticides on water bodies, in which the test object is kept in the test solution, record the survival rates of the test object upon which to calculate the threshold concentration that is lichnosti the tested pesticides, and assess the degree of toxicity of pesticides, characterized in that it further register pathological changes in the test object, calculate the average percentage of deformities and as the threshold concentration teratogenic effects set the concentration of the pesticide, which has less teratogenic effect on the test objects, and on the degree of toxicity of pesticides is judged on the basis of the coefficient of threshold concentrations, which are calculated according to the formula

where Kpemb- coefficient of threshold concentrations the toxicity of pesticides, taking into account their teratogenicity;
LC16- threshold concentration of the toxicity of pesticides, causing the death of 16% of embryos;
ETerK16threshold concentration teratogenic effects of pesticides,
when the values of Kpemb
>10 establish the hazard class of pesticides I, i.e. extremely dangerous.
5-10 installing the hazard class of pesticides II, i.e. highly hazardous;
1-5 installing the hazard class of pesticides III, i.e. dangerous;
<hazard class 1 pesticides IV, i.e., moderately dangerous.

2. The method according to claim 1, characterized in that the test object using embryos sturgeon.



 

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3 cl, 7 dwg

FIELD: processing procedures.

SUBSTANCE: invention can be used in analytic chemistry for sorption concentration and successive determination of heavy metals in water solutions. The procedure for production of sorption material consists in impregnation of surface of a cellulose filter with an analytic reagent wherein thio-semi-carbazone of picoline aldehyde is used as such. Impregnation is carried out with conditioning cellulose material in solution of the reagent in ethanol containing 2.5 % of cetyl alcohol with successive extraction and drying in air. Produced cellulose material is applied for sorption-roentgen-fluorescent analytic determination of heavy metals in water solutions. Metals are extracted with cellulose material for roentgen-fluorescent determination at pH 7.5-10.5, preferably, at pH 10.0.

EFFECT: simple and safe procedure for production of sorption cellulose material used for efficient concentration of heavy metals with successive determination of each of them separately and in whole.

4 cl, 2 tbl, 2 ex

FIELD: chemistry.

SUBSTANCE: in order to realise the method, aromatic amines are extracted from waste water with an emulsion of aqueous solution of an inorganic acid with ionisation constant higher than 10 in an organic solvent.

EFFECT: high degree of extraction of aromatic amines from waste water with minimum consumption of reagents and combining extraction and re-extraction processes at one step.

3 ex

FIELD: chemistry.

SUBSTANCE: method of determining crystallisation of heavy isotope types of water during volumetric, uniform cooling of natural water and formation of ice of heavy water involves determining and recording changes in optical properties of water using a laser beam and two photocells. The photocells are placed at different heights and the laser beam and its scattered radiation are picked up. The laser beam is pulsed with pulse duration of up to 1 second and period between pulses of 30-200 seconds. Measurements are taken after lowering temperature of the processed water to +4°C. Before each measurement, the water aeration process is stopped completely or only on the area under the beam for the period of time when bubbles surface.

EFFECT: invention increases quality of water and preserves its salt composition.

1 dwg

FIELD: medicine.

SUBSTANCE: nanobacteria are counted in a human nephrolith. A fixed mass is separated from the latter, mechanically powdered and divided into j=5 weight fractions pj. The powder is poured into j=5 sterile cells, water infiltrate at pore size not exceeding 0.05 mcm is added. The concentrations of nanobacteria is set between 102 to 106 cells in 1 ml by varying the water volume Vj or weight fractions pj of a powder mineral mass in each cell with using the formula. It is followed with mixing poured into j measuring cells. A nutrient medium - calves' fetal serum is added in the ratio 1:9. Two electrodes are inserted in each cell, then the measuring cells with the mixture is placed in an autoclave wherein constant temperature within 30°C≤T≤40°C is maintained. A mixture impedance (R) is periodically measured, and a point of measurement time (t) is determined until a mixture impedance slump is observed. A calibration diagram of an impedance variation time (timpj) to the concentration of nanobacteria in an initial sample (timpj) is presented. Thereafter, the above-stated stages of the method are conducted for analysed water as well. The derived impedance time (timpj) values are projected on the calibration diagram on the axis (timpj), then on the axis (lgnj).

EFFECT: invention allows evaluating the water concentration of nanobacteria.

3 dwg, 1 ex

FIELD: chemistry.

SUBSTANCE: method involves predicting composition of a nonvariant solution, experimental determination of compositions of mixtures on boundaries of the nonvariant region and nonvariant liquid phases with optimum initial mixtures of components, lying in a strictly defined order, from measurements of the physical property of the liquid phase after establishing equilibrium using the "composition-property" functional relationship. Further, for all experimental points lying arbitrarily on all boundaries of the nonvariant region, based on the average ratios of content of the component which is absent in compositions of equilibrium solid phases on corresponding planes or extreme nodes of the nonvariant region of the system to content of water, the composition of the nonvariant liquid phase is calculated and equilibrium solid phases are determined using true coordinates.

EFFECT: obtaining accurate data on phase equilibria in a system without using chemical analysis methods, considerable increase in accuracy mathematical prediction of the composition of the nonvariant solution, avoiding loss of accuracy and reliability of determining composition of the nonvariant solution with increase in the number of components in systems of any type.

4 tbl, 3 dwg, 2 ex

FIELD: chemistry.

SUBSTANCE: in the method for rapid detection of heavy metals in water, a biosubstrate is mixed with a solution containing heavy metals, wherein the solvent used is ethyl alcohol and water in ratio of 6:1, and the structure index of crystallograms obtained in the presence of salts of heavy metals Cu2+, Fe2+, Zn2+ is determined.

EFFECT: simpler and faster analysis.

3 dwg

FIELD: analytical methods in environmental monitoring.

SUBSTANCE: method comprises: sampling, acidifying samples with HCl/H2SO4 mixture, adding Ce(SO4)2 as oxidant and removing its excess with reducing agent NH2OH·HCl, adding rhodamine C as organic reagent, extracting resulting complex, separating organic phase from aqueous phase, and measuring optical density. Extraction is performed with carbon tetrachloride/methyl isobutyl ketone mixture at 5:1 volume ratio and extractant-to-sample volume ratio 1:1 under dynamic conditions by way of washing away complex with extractant. Content of antimony is judged of from difference of optical densities of extractant and mixture.

EFFECT: lowered measurement threshold to values comparable with allowable limits, increased reliability, reduced analysis time, and automated analytical procedure.

3 cl, 1 dwg, 3 tbl, 3 ex

FIELD: environmental monitoring.

SUBSTANCE: invention relates to hygiene and sanitary of freshwater reservoirs and is meant to be used for microbiological testing of condition of water source in an agricultural enterprise effluent zone, in particular in the effluent zone of poultry factories and pig-breeding farms. For this aim, water is sampled at least in two zones: in the effluent zone of agricultural enterprise and in the zone, where influence of agricultural enterprise effluent over different periods is excluded. Then, contents of ammonia compounds and enzymatic activities of urease-carbamidamidohydrolase in samples are measured. Obtained data are processed: contents of ammonia compounds and enzymatic activities in different samples are compared to each other and dynamic of changes in data in samples taken from different zones are compared.

EFFECT: enabled quick testing at high accuracy in estimations and minimized labor and means involved.

2 tbl

FIELD: toxicology, in particular determination of water flea sensibility to toxic effect of water-soluble chemicals.

SUBSTANCE: claimed method includes detection of water flea death time (min) caused by water-soluble chemicals, wherein concentration (C, mol/l) of chemical under consideration fluctuates according to logarithmic scale with interval of 0.1. Plot of Y versus X is made, wherein Y-axis represents average death time with scale of 1 point = 1 min; X-axis represents reverse concentration (1/C) of chemical under consideration; and scale is proportional to log increasing by 0.1. Water flea sensibility to toxic effect (tgα) is calculated according to equation: tgα = TL(min):1/KL = TL(min)xKl (I), wherein α is inclination of straight line to X-axis; TL(min) is death time (min) being determined according to point of hypothetical crosspoint of straight line with Y-axis; KL is lethality constant (mol/l) defined as chemical concentration wherein water flea death time is equal to 2TL(min).

EFFECT: Method allowing evaluation of toxic effect evolution dynamics and comparison of toxic effect of water-soluble chemicals in equal concentration ranges.

2 tbl, 1 ex, 1 dwg

FIELD: analytical methods in industrial sanitation.

SUBSTANCE: method envisages bringing solution to be analyzed into contact with potassium bichromate, sulfuric acid, and mercuric sulfate, ageing resulting mixture and allowing it to cool to ambient temperature, adding ferroin indicator, titration of excess of potassium bichromate with 0.125 n. Moor salt solution, and calculating chemical oxygen demand value from amount of Moor salt consumed in the titration. Method is characterized by that initial solution is preliminarily homogenized until diameter of suspended particles therein become as large as 0.03 mm, after which solution is allowed to stay for 4 min.

EFFECT: reduced determination inaccuracy.

FIELD: environmental protection.

SUBSTANCE: invention concerns estimation of environmental pollution using bioassay methods. In particular, method is accomplished through bioindication of controlled area using, as bioindicators, internals (muscles, kidneys, liver) of wild hoofed animals (elk, dear, wild boar). One determines content of heavy metals in these organs placed within an area, compares thus obtained data with maximum permissible concentrations of heavy metals in foods, and estimates heavy metal pollution level of the area from resulted difference. Existence of long-term pollution of a region is judged of from excess concentration of heavy metals in wild hoofed animal kidneys and existence of single release of mercury and lead from that in muscles and liver.

EFFECT: enabled multiple estimation of considerable areas at reduced effort.

2 cl, 3 tbl

FIELD: analytical methods in environmental protection and toxicology.

SUBSTANCE: subject of invention is drinking, natural, and waste water quality monitoring. Toxicity of aqueous medium is determined from variation in activity of animal brain plasma membrane Mg2+-ATPase activated by chlorine and/or bicarbonate ions. In one embodiment of invention, toxicity of aqueous medium is determined by mixing above plasma membranes with test aqueous medium adjusted to physiologic pH with phosphorus-free buffer followed by addition of solution containing Tris-ATP and magnesium ion source as well as chloride and/or bicarbonate ion(s) source, incubation until inorganic phosphorus is formed, and determination of toxicity from concentration of inorganic phosphorus. Brain plasma membranes used as indicator contain Mg2+-ATPase capable of being activated by chlorine and/or bicarbonate ions.

EFFECT: extended functional possibilities of method and use of reagent, increased sensitivity, and enabled determination of toxicity at lower concentration of various-type toxicants.

20 cl, 6 tbl

FIELD: environmental protection.

SUBSTANCE: invention concerns evaluation of pollution of areas with pesticides involving bioassay techniques. To that end, area under control is subjected to bioindication using wild hoofed animals (mainly elks, deer, wild boars) as bioindicators. Within specified period of time, animal internals are sampled, pesticide content therein is determined, and thus obtained results are compared with maximum permissible pesticide levels for food products. Comparison data are used to estimate quality of media.

EFFECT: increased representativeness of monitoring results, enabled evaluation of a vast region or local area or local agrocenosis at lower effort.

2 cl, 2 tbl

FIELD: chemistry, water quality control, method for quantitative estimation of organic substance properties in aqueous solutions.

SUBSTANCE: indicator plate is immersed in aqueous solution and according to alteration of aqueous solution composition chemical activity of organic substances in this solution is determined. In clamed method tree vessels are used. Two vessels contain aqueous solution to be tested and the third vessel contains control aqueous solution free from organic contamination. In aqueous solutions containing in the second and third vessels indicator plates are immersed, then aqueous solutions in all vessels are heated up to 95-105°C, held at this temperature for 55-65 min, cooled to 15-25°C, filtered though membrane filter with pore size of 0.46 mum, then iron content is measured in all vessels and chemical activity of organic substances in aqueous solutions is calculated according to equation ka=ΔFe/Fe1, wherein ΔFe = Fe2- Fe1- Fe3; Fe1 is iron content in the aqueous solution of the first vessel; Fe2 is iron content in the tested aqueous solution of the second vessel; and Fe3 is iron content in the aqueous solution of the third vessel.

EFFECT: simplified method with enhanced functionality.

3 ex, 4 tbl

FIELD: ecological engineering, particularly river monitoring with taking into consideration river pollution with sewage water within the limits of cities and other inhabited localities.

SUBSTANCE: method involves choosing river observation points relative single pollution source or pollution source array; aligning one observation point with single pollution cross-section or pollution cross-section array; taking water samples; conservation the samples and preparing thereof for following analysis; cultivating test-organism, namely one-celled green algae - Chlorella vulgaris, inside cultivator at temperature of 36±0.5°; measuring optical density thereof in red light; analyzing and estimating the measurement results. The observation points are transversal to river and set in front of single sewage pollution source or sewage pollution source array, in center of each source and behind them. All three observation points are located within the limits of a city or other inhabited locality. The optical density is measured before and after one-celled green algae cultivation in water samples. After measurement termination overall river water pollution index is determined.

EFFECT: possibility to compare overall river water pollution index obtained in particular observation point with that obtained from pollution source on river bank; extended functional capabilities and extended range of application.

5 cl, 4 tbl

FIELD: ecology.

SUBSTANCE: the present innovation deals with testing biological activity of water, preliminary treatment of water, division into control and tested portions, ionization of tested portion with silver ions, detection of the quantity of sprouted wheat grains per time unit pre-impregnated in both mentioned portions and calculation of relative alteration for the value of biological activity of water according to the following ratio: where d - relative alteration for the value of biological activity of water, %; Ntested - the quantity of sprouted grains per time unit in tested portion of water, pcs.; Ncontrol - the quantity of sprouted grains per time unit in control portion of water, pcs. Moreover, relative alteration for the value of biological activity of water being above 0 means increased biological activity of water, relative alteration for the value of biological activity of water being below 0 means its decrease, and equation of the mentioned relative alteration to 0 means intact nature of biological activity of water being different by the fact that pre-treatment of water should be carried out due to precipitation for 23-24 h at 23-26 C.

EFFECT: higher efficiency of investigation.

1 tbl

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