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B1. Title page
Proposal full title: Development of Methodology of Express Evaluation and
Express Evaluation of Genotoxicity of Environmental Pollution Chemicals.
Proposal acronym: DMEEEEGEPC
Date: 05.10.99
B2. Table of Contents (main, headings, page number):
B1.Title page............................................................................................ 1
B2.Table f Contents.................................................................................. 1
B3.Objectives and expected achievements............................................... 1
B4.Contribution to programme/specific action objectives....................... 2
B5. Innovation aspects.............................................................................. 2
B6. Project work plane.............................................................................. 3
a) Introduction..................................................................................... 3
b) Project planing and time time table................................................ 5
c) Graphical presentation of project’s components: (Pert diagram).. 6
d) Detailed project description broken down into work packages:
1) Work package list (form WP1)................................................ 6
2) Deliverables list (form DL)...................................................... 8
3) Form WP2................................................................................. 9
B3. Objectives and expected achievements
Purpose of this project is, using with developed metodology, to give an
express-evaluation potential
genotoxicity of chemicals, which pollute
atmospheric air and drinking water in the given region.
Objects of studies will be:
à) samples of atmospheric air from large industries cities of region;
b) atmospheric air along large autobans with active car traffic;
c) exhaust
gases of different
diesel cars (for the comparison of
genotoxicity directly diesel exhausts and general gas-ladenity of streets);
d) samples of drinking water from city water-pipes with the different
technology of water preparation (using as desinfectants such oxidants,
as
chlorine, dioxide of chlorine, ozone, chloroamyne on
initial and end stages of
water preparation);
As a test-objects will be used microorganisms Salmonella typhimurium,
adapted for genotoxicity studying of chemicals by Ames and co-authors.
The half-quantitative Ames test with metabolic activation will be applied for the
express-investigation of chemical genotoxicity. It was adapted by us for testing
the genotoxicity of complex chemicals mixes, extracted from environment
(atmospheric air and drinking water ).
As a result of conducted studies will be:
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à) given ecogenetics evaluation of state of atmospheric air from large
industrial cities;
b) on results of biotesting the samples of air and water on potential
genotoxicity given a conclusion about potential ability (or inability) chemicals,
identified from under investigation environmental objects to induce mutations in
sex and somatic cells of human population, which lives in the given region;
c) physical and chemical analysis of samples of air and water will allow to
identify chemicals classes and groups which induce the genotoxicity effects
inside the human population, living in the given region. That will enable to
develop corresponding legal documents on the admission (or, at least once,
restriction) getting these chemical in the to surroundings;
d) realization of points a), b) and c) present studies will reduce a frequency
of hereditary and oncology diseases of regions;
e) using in this project offered by us method of evaluation genotoxicity of
surround chemicals reduce the price of determined purpose realization on 50 %.
В4. Contribution to programme/specific action objectives
Practical executartion of this project will consist of 4-h components:
1) selection of samples of air in different parts of the city; samples selection
of drinking water on initial and end stages of water preparation and on plumings,
using different oxidants for water desinfection;
2) preparing the select samples to study the genotoxicies characteristics and
undertaking physical and chemical analysis: concentrating, identification class
and groups of chemicals;
3) physical and chemical analysis of air and water samples on the contents in
them chemicals, which are able to induce genome, chromosome and gene
mutations, that can bring to increasing a frequency of hereditary and oncological
pathology;
4) biotesting of prepared air and water samples on the ability to induce
mutations by complex of the chemicals from these samples;
5) the attempt will be made to calculate the genetical risk of heriditary
pathology in the region using projects materials (and the literature data
concerning the heriditary pathology frequency in the region) and in connection
with contamination of environment by mutagenic chemicals.
В5. Innovation aspects
Realization the proposed project for the first time for countries of
Euroalliance will give:
a) strategy and tactician of ecological and genetical studies, have been
directed to the evaluation is qualitative and quantitative state of environmental
objects (atmospheric air, water), which is a component part of genetic
monitoring;
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b) the ecological and genetical
feature of environmental objects
(atmospheric air and drinking water);
c) physical and chemical analysis of air and water will allow to reveal
classes, groups and separate chemicals, are responsible for genotoxic activity of
the whole sample. It will promote a deciding about in admission (or limitation) of
getting these materials in the environmental;
d) ability (or inability) of chemicals complex, which were picked from the
definite volume of atmospheric air and water to show genotoxical characteristics
by inducing gene, chromosome and genomic mutations;
e) presence of the mutagen activity in a select samples of water and air and
chemical analysis data will be a base of prognosis calculations of increasing a
hereditary pathology frequency in the region.
В6. Project work plan:
a) Introduction.
In last 30-40 years in consequence of speed industry development, intensive
using the chemicals in agriculture, appearance a new medicine and home
chemicals, a lot of dangerous chemical materials get inside environmental
objects (atmospheric air shallow and underground reservior, soil and foodstuffs). Many from them are able to influence on heredity material.
Chemical mutagens action on human heredity is result of increasing a
frequency diseases (due to mutations, which were induced in parents sex cells0
or development malignancies (due to mutations, which were induced in somatic
cells). The spetial danger of chemicals with mutagenical capabilities is they may
be a cause a significant increasing a recessive mutation frequency. It leads to
heavy heriditary diseases, which are not show in the first generations, they may
be a cause of sick rate «hits», of radually accumulating in populations.
At the present time the questions of chemical mutagenesis are studied so
intensive in many world laboratories. As result, evident mutagenical action of
many chemicals has been shown. Quantity of such investigations increases
quickly and at the same time quantity of chemicals with described characteristics
increases also.
It is offered about 100 different test-system for assessment of chemicals
genotoxiciological characteristics. Most of all are elaborated so much in the
metodological aspect and used bradly as for mutagens revealing from enormous
quantity of chemicals (methods of screening) as evaluation their genetic danger
for human. However universal strategy does not exist, which could reveal all
main types of genetic damages. This fact dictates need using in the work the
collection of methods, which allows to discovery and evaluate the mutagenicity
of investigated agents. So, different types of organisms serves as a test-objects:
from bacteriofages and viruses to mammalian and human cells in conditions in
vitro and in vivo. During last years it has been made a lot in aspect evaluation of
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aplicability offered methods for screening and chemicals mutagenical potentials
investigation. Main criterious of evaluation were working out degree and scales
of using, realization speed, cost and possibility application this method to
practice. Big quantity of contaminators, which have olready been in different
environmental objects (in the atmospheric air and houses air, different types of
water, in food stuffs), and transformation phenomenon of them inside these
objects lead to traditional analysis methods can not conclude about real
genotoxicological contamination of these objects. It forces to search new
approaches to real assessment of genotoxicity of environmental contaminator’s
complex.
One of such approaches can serve a method of testing the samples from
environmental objects have been tested directly in the biological test-systems
after inducting the definite selection and preparation. This approach named by
us in future as evaluation of total mutagen activity (TÌÀ) environmental objects,
allows firstly to value real genotoxicity of environmental object wholly and,
secondly, if activity becomes to be increased - to conduct a further study of such
contaminators by the way fractionation with the following evaluation of separate
factions and using the chemical analysis. It finally, will allow to reveal separate
materials, which stimulate of genoroxicity the whole sample.
Study of total genotoxicity of chemical contaminators of air and water
objects are actual also because qualitative state of atmospheric air, drinking water
and food-stuffs (according to factors of total genotoxicity) can be a reason of
increasing a hereditary pathology frequency. Known that lung cancer to a
marked degree depends on presence in the inhaled the air mutagen-active
(carcinogenic) chemicals. In the process of water preparation on stages of
primary chlorination enourmous quantity of chemicals, which contain Cl, are
formed. Most of all have potential mutagen and carcinogenic characteristics.
Detailed mutagenic study of complex of environmental contaminators with
aim to exclude or (at list)limit their contact with human population is capable to
reduce a spontaneus abortion frequency to some extract, that, in the end could
improve the genetic health of populations from Euroalliance.
Planned work will be realised by four directions:
a) atmospheric air and drinking water samples selection;
b) preparing the samples of water and air for studies (eluation,
fractionation);
c) physical and chemical analysis of samples on presence in them
genotoxical chemicals;
d) realization the experimental studies of total genotoxicity atmospheric air
and water samples;
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b) Project planning and calendar plan. This project is planed to execute 2
year.
The first year of studies will be dedicated to analysis of total genotoxicity of
atmospheric air in different points of city and in different places of active
superhighways. As a result of annual work will be formed a map of city with the
indicating of the most polluted by genotoxic chemicals regions and streets.
The second year will be dedicated to analysis of the total genotoxicity
samples of water from of two water stations. Mutagenicity of initial water and
drinking water, passed trough different technological ways of water preparation
(initial ozonation, initial chlorination), will be studied.
Calendar plan will look as follows.
Calendar plan of work
N
1.
2.2.
3.
Name of stage of work
Terms of execution
2000
a) Selection of the atmospheric air samples in
Weekly during 2000
3
the city (in 10 points weekly, 100 ì in each
point);
b) Selection of air samples on active city
Weekly during 2000
3
pathways (weekly, 100 m in each spot)
Preparing of air samples to the further
As far as the
studies (eluation, fractionation)
samples will be arrived
Execution a physical and chemical analysis of air As far as the
samples on presence the genotoxical chemicals in side
samples will be arrived
them
4.
Study of total genotoxicity of air samples
5.
Writing the annual report
1.
2.
3.
4.
5.
2001
Selection of samples from two water-supply
and drinking water
Preparing the water samples to further studies
As far as the samples
will be arrived
November-December
Monthly
As far as the samples
will be arrived
On measure of arrival
of samples
Execution physical and chemical analysis of
water samples on presence the genotoxical
chemicals inside them
Study of total genotoxicity of samples of water As far as the samples
will be arrived
Writing the final report
November-December
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c) graphic presentation of components of project (Pert diagram);
WP1
WP2
WP3
WP4
а)
б)
в)
a)
б)
а)
б)
а)
б)
Mont
h
3
6
9
12
15
18
21
24
d) The Detailed project description divide working packages into
(stages)
WP1.
Work
package
(этап) No
WP1.
WP2.
WP3.
Work package list
End
month
Phase
Deliverable(s)
January,
2000
December,
2000
«RS»
D1.
January,
2000
December,
2000
1 .
January,
2001
а) Preparing of air samples to
further studies;
2
January,
2000
December,
2001
December,
2000
«RS»
D2.
b) Preparing of water samples
to further studies
2
January,
2001
а) Execution physical and
chemical analysis of air samples
2
January,
2000
«RS»
D3.
a ) Selection of air samples in
city (in 10 points weekly, 100м3
in each point);
b) Selection of air samples on
active city pathways (weekly
100м3 in each point);
c) Selection of water samples
on two water stations of city
and drinking water.
Respon
sible
partici
pant
No
Person-
Start
month
Work package title
months
1
1
42
42
December,
2001
December,
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on
presence
genotoxical
chemicals in side them;
WP4.
2000
b) Execution physical and
chemical analysis of water
samples
on presence
genotoxical chemicals in side
them;
2
а) Study and analysis of total
genotoxicity of the air samples;
1
b) Study and analysis of total
genotoxicity of the wate
samples
42
January,
2001
December,
2001
January,
2000
December,
2000
December,
2001
42
1
January,
2001
«RS»
D4.
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DL
Deliverable N
DL1.
DL2.
DL3.
DL4.
D5.
D6.
D7.
D8.
Deliverable list
Deliverable title
a) Selection of the atmospheric air
samples (in 10 points weekly,
100м3 in each point;
b) Selection of the air samples of air
on active city pathways of city
(weekly, 100м3 in each spot)
Preparing the of air samples to
further studies
Execution a physical and chemical
analysis of samples air on presence
genotoxical chemicals inside them
Study of total genotoxicity of air
samples
Selection of water samples from
two water-supply and drinking water
Preparing the water samples to
further studies
Execution a physical and chemical
analysis of water samples
on
presence genotoxical chemicals
inside them
Study of total genotoxicity of water
samples
Delivery
date
Nature
Dissemination level
R
PU
R
PU
R
PU
R
PU
R
PU
R
PU
R
PU
R
PU
R
PU
31.12.00
31.12.00
31.12.00
31.12.00
31.12.00
31.12.01
31.12.01
31.12.01
31.12.01
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Work package description
Work package number: WP1;
Start date or starting event: January, 2000;
No of the partner responsible: 1;
Nos of other partners involved 1
Person-months per partner 42
Objectives: WP1: Selection of the air samples, selections of water samples from river and
drinking water samples from two water- supply will be realize for ensuring by the scientific
material:
а) physical and chemical analysis of contents an genotoxicants in the samples;
b) study of genotoxicity of chemical contaminations in the water and air samples
Description of work. The atmospheric air samples will be select weekly on stationary posts of
monitoring for on glass filters in different points of city. Volume of selected air in one point is
100м3. Filters will be united and overed monthly sample will be subject for physical and chemical
analysis and biotesting on total genotoxicity. Samples of water will be selected in the river region
and from the reservior of clean water (drinking water) on two water stations (on one of them as
secondary desinfectants ozone is used instead of chlorine), which supply large city by drinking
water.
Volume of one water samples
will form 30 l. Multiplicity of concentrating of the
chemicals will form х10000 once.
Deliverables. As a result of carrying this stage of work two short information reports about
samples selection of atmospheric air and water will be direct out to in the scientific sector an
Ukraine-austrian institute
Milestones and expected results. Execution of WP1 work-stage is planned to realize during two
years. In 2000 atmospheric air samples and samples of air from active city superhighways will be
selected weekly. Reports about executed work will be afford to management of Institute quarterly
(more aften or less depending on requirements). Within 2001 water samples from river and
drinking water samples from two water stations, which use different technologies of water
preparation (different desinfectants), will be planned to select monthly. Reports about executed
work will also be afford in accordance with requirements of Institute. To expected results of allows
to refer the following: as far as WP1 stage.
Is only technical, the result of this work - an quantity of a select samples of atmospheric air:
10 stationary posts x 12 months = 120 samples; quantity of a select of water samples - 2 water
stations x 2 (river and drinking water) x 12 months = 48 samples. As far as the purpose of project
is conclude in studing the genotoxicity of chemicals water and air contaminators within a year,
consequently, selections of water and air samples - processes are unceasing (so long keeping of
samples in admissible because of chemicals destruction), but signifies, clearly outlined stages of
this work does not exist.
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Workpackage description
Work package number: WP2;
Start date or starting event: January, 2000;
No of the partner responsible: 2;
Nos of other partners involved 2
Person-months per partner 42
Objectives. WP2: Preparing of the water and air samples to studies, i.e. concentrating all
chemicals in volumes, suitable for chemical analysis and studying of chemicals genotoxicity,
identified from water and air.
Description of work. Weekly selected air samples are united and average-month sample is formed
on each point of selection. This sample is subjected to a processing in the device of Sokslet by
organic solvents so, that maximum quantity of chemicals moves over to dissolve. Then solvent is
got in the sweat, but dry remainder is fractionated for the determination of chemical composition
and is dissolved in dimetylsulfoxyde (DMSO) for genotoxicity studying.
The water samples also are subjected to processes of concentrating the chemicals in small
volumes by way of organic extruction of chemicals, evaporating of organic solvents, fractionation
for the determination of chemical composition dissolutions of the dry remainder in DMSO for
genotoxicity studying. All practical work of preparing the water and air samples to studies is
planned to conduct on the base the labs. of hygenic studying the carcinogenic factors.
Deliverables. As a result of execution this stage of work two reports will be written and present in
the scientific sector an Ukraine-austrian Institute: about preparing the atmospheric air and samples
about preparing the water samples.
Milestones and expected results. The working at present stage is planned to conduct during
two years on the measure of amples sarrival. In 2000, 120 atmospheric air samples will be prepare
to the study and undertaking a physical and chemical analysis studying of genotoxicity. 2001 years
will be denoted to similar preparation 48 water samples to the same studies. The work of
preparing the samples to the study will be hold at the measure of their arrival. As far as the
purpose of project is conclude in studing the genotoxicity of chemical water and air pollutant
within a year, consequently, and selections of water and air samples and preparation them to
further studies - processes are without pause (i.e. long keeping of samples inadmissible because of
chemicals destruction, but, clearly outlined stages of this work, and in this case, does not exist.
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Workpackage description
Work package number: WP3;
Start date or starting event: January, 2000;
No of the partner responsible: 2;
Nos of other partners involved 2
Person-months per partner 42
Objectives. WP3: Execution a phisical and chemical analysis of atmospheric air and water
samples will be conduct for the reason to identificate (as far as it is posible) maximum quantity of
chemicals classes and groups responsible for total genotoxicity of sample.
Description of work As far as the atmospheric air and water samples will be arrived after
preparating by special way, they will be tested by physical and chemical analysis for cheking on
content the main classes and groups of chemicals, which may induce the mutation, inside them
(samples). This work will be conduct on base a labs. of carcinogenic factors.
Deliverables. As a result of performing this stage of work will be written and sent in the scientific
sector an Ukraine-austrian Institute two reports with the detailed enumeration of strategies of
analysis and received concentrations of chemicals, in the calculation on the cubic metre for the
atmospheric air and mg/l for drinking water.
Milestones and expected results. The working a present stage is plan to conduct during two years
on the measure of arrival of tests. At 2000 years will be analyse 120 tests of atmospheric air and is
determined contents in them of genotoxic chemicals. 2001 years will be denote similar analysises
of contents of genotoxicants in 48 tests of water.
The work on the physical and chemical analysis will be hold at the measure of their arrival.
As far as purpose of project is conclude in that, to study of genotoxicity of chemical soiling water
and air within a year, consequently, and selections of tests of water and air, and preparation them to
further studies, and physical and hemical analysis of chemicals - processes unceasing (i.e. long
keeping of tests inadmissible because of destruction of chemicals), but signifies, clearly outlined
stages of this work and in this case does not exist.
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Workpackage description
Workpackage number: WP4.
Start date or start event: January, 2000
N of the partner responsible: 1
Nos of other partners involved 1
Person-months per partner 42
Objectives. WP4: Study and evaluation of total genotoxicity samples of water and air will be made
for the forecast development of hereditary diseases of population of city in connection with soiling
of environmental (approach to the given forecast development can be realize for other regions).
Description of work Working a given stage is also calculated for 2 year. At 2000 years is plan
study and evaluation total genotoxicity complex of chemicals, identify from the air.
For these whole will be used the most popular in the world a strategy Ames, adapted by us for
studying mutagenecity complex mixtures with offered us mathematical way of processing primary
data.
Within 2001 year will be conduct evaluation total genotocxicity samples river and drinking
water of two water stations. On material a physical and chemical analysis biotesting the complex
mixtures on total genotoxocoty and on literature given on hereditary pathology frequency will be
made attempt to give forecast of genetic risk for population EU in connection with soiling a
environmental genotoxical chemicals
Deliverables As a result of do work will be present report with calculation of forecast of risk of
increasing a hereditary pathology frequency in connection with soiling of environment.
Milestones and expected results The Working a given stage will also be conduct during two years
on the measure of arrival of samples. Received results on genotoxicity of complex of chemicals
from environment, on contents of mutagenic chemical pollutants, as well as (from literature
sources) on hereditary pathology frequency will be calculated genetic manifestation risk of
hereditary diseases in connection with pollution of environment be chemical genotoxicants.