DISCOVERING GENOTOXICITY ON COW FARMS IN THE AREAS WHERE
HARMFUL MATTERS WERE PRESENT DUE TO PETROL BURNING
Slavica Kosarcic*, D. Kosarcic, Mira Kovacevic
• Scientific Veterinary Indtitute Novi Sad, Rumenacki put 6, Novi Sad, YU;
tel.: ++381 21 311 066; fax: ++381 21 317 544; e-mail: [email protected]
ABSTRACT
It is very well known and experimentaly proved, that harmful biotic and abiotic
factors influence genetics of the living beings. They cause numerical and structural
changes in chromosomes. The aim of our investigation was to discover, by
cytogenetic methods, if there are some damages on chromosomes in the cows that
were in the surrounding of a petrol refinery where enormous quantity of harmful
matters was released.
Thirty cows were chosen on a farm, 5 km away from Novi Sad refinery, where for
many days harmful matters, soot and other toxic gases were present. The other
group of the same number, was 70 km away from the source of toxication. The
control group of 60 cows were analyzed last year. Cytogenetic analyses was done
according to the modified method of Moorhead et al., Seabright et al. and according
to international standards for karyotypisation of animals.
In the first experimental group 3 animals were discovered with the changes of break
in q-arm and aneuploidia. The other experimental group and the control one had
good cytogenetic finding 2n=60XX.
According to the results it can be concluded that there are genotoxic matters, that
appeared in nature through burning of petrol in the refinery, as a consequence of
bombing, and they influenced structural changes on chromosomes. Further
investigation will be done on the animal's offspring where aberation was detected.
Keywords: cows, karyotype, changes, genotoxic.
INTRODUCTION
Observing changes in the living beings that can be a consequence of harmful
influence on the surrounding, demands accepting the fact that the ecological areas
cannot be equated with geographical areas of the world. Therefore, it is very
important to create a general opinion in every person that distorting biological
balance and law in matter circulation is refleced all over the world, no matter where it
happenes. It is true that the influence of abiotic and biotic factors had greater impact
on the living beings and Eco-system in whole.
The aim of our investigation was to check if there was a harmful influence on
the cow genome caused by toxic matters that were present in the air, water and feed
due to incomplete burning of petrol. Namely, during the air raids on FR Yugoslavia in
the period from March 24 to June 6 1999 the refinery in Novi Sad was bombed. On
this occasion on the processing and the storage equipment 270 missile were thrown
on the total of 73,770 tons of petrol and its derivates. Quantity of 65,180 tons burned
and the rest was poured in the Danube and the surrounding area. According to the
information of the Federal Ministry for Development, Science and Environment
enormous quantity of harmful matters, toxic gases and soot were released.
Concentration of sulfur-dioxide (SO2), carbonmonoxide (CO) and soot in the area
were several time higher than permitted. (For example concentration of SO2 in the
city was 580µg/m3.) In the underground water and soil were present the following
harmful matters: lead, all oxidohydrogens, polycyclic aromatic oxidohydrogens and
polychlor bifenoles.
The samples were taken on 40 places around the refinery. The air, water and the
soil were analyzed on the cow farm where we examined genotoxicity. According to
the results from this area, the quantity of the harmful matter present in the water, air
and soil were above the permitted values. This points to the fact that percentage of
the toxic matters due to incomplete burning of petrol entered the chain of
nourishment and cannot be controlled. Above mentioned chemicals are considered
harmful for DNA molecules, the carriers of inheritance in almost all living beings on
the earth (3, 4). Therefore, we decided to examine genotoxicity on the cow farm that
was in the area of air raids.
MATERIAL AND METHODS
For examining the harmful matters in the animal gene, 120 cows were chosen
on two different localities. The first group of 30 cows was on a farm 5 km away from
the refinery where during the bombing enormous quantity of toxic gases and soot
were released. By chemical analyses of food, water and air genotoxic matters were
discovered. The other group, that consists of 30 cows, was randomly chosen on a
farm 70 km away from this area. A control group consists of 60 cows, 30 cows on
each farm, and were analyzed last year. From the experimental animals by a sterile
procedure blood was taken from the jugularis vein and the lymphocyte culture was
established according to the modified method of Moorhead et al. The analyses of
the chromosome in all the animals was done according to the international
standards for caryotypisation in the domestic animals ISCNDA 1989.
RESULTS AND DISCUSSION
After examining the karyotype, 100 metaphase were done for every animal
and here are the following results:
Control group had clean cytogenetic finding 2n=60XX (Figure 1).
Figure 1. Microphotography of a normal karyotype in
the cow from the first control group, 2n=60XX, Geims
strain 1000X.
In the first group that was for many days exposed to harmful matters, 3 cows
were discovered with numerical and structural changes in the karyotype. These
changes on the chromosome were the kind of aneuploidya 2n>60 and the structural
change of type brake on q-arm in X chromosome and autosome. The animals with
aneuploidya in its karyotype carried two kinds of cells and it was 11% with higher
number of the chromosomes 2n>60, 2n=89 and 23% cells with 2n>60, 2n= 73,
figure 2 and 3.
Figure 2. Microphotography of karyotype with aneuploidya 2n>60XX, 2n>89XX
Geims strain 1000X
Figure 3. Microphotography of karyotype
with aneuplodya 2n>60XX, 2n=73 Geims
strain 1000
Structural changes with a break on both X chromosomes were discovered in
the cow where miscarriage occurred in the first half of gravity. Analyzing the
karyotype with the mentioned structural changes in the genome in 100 metaphases,
49% of the cells had a brake on q-arm of one chromate on both X chromosomes,
Figure 4.
Figure 4. Microphotography of karyotype with break on q-arm X chromosome, Geims
strain 1000X
Next structural change, type brake on q-arm of autozome No. 5, was
isochromatide and was discovered on one chromosome in homologue pair, Figure 5.
Out of 100 examined metaphases these chromosome distortions were discovered in
13% of the cells. The cow with the changes had no obvious problems in health
condition and reproduction.
Figure
5.
Microphotography
of
karyotype with isochromatide brake on
q-arm, a chromosome in the fifth pair.
Giemsa G-banding 1000X.
After cytogenetic analyses of the second group of cows, 70 km away from the
area of toxicating, cytogenetic findings were good 2n=60XX.
Discovered changes in the genome of the endangered animals point on certain
genotoxicity of agens from burned petrol but there are no sure evidence on targeted
influence. This requires further investigation and introducing accurate methods in
water, food and air control and parallel investigation of the discovered agens in
tissues. The results from numeric and structural changes in karyotype indicate that it
has noting to do with constitutional karyotype that is present in all the cells [ 1], but
with damages on chromosomes that appeared de novo, most probably as a result of
some genotoxic matters [5,6]. Available data from literature and experimental proves
in large number of the investigations, make us conclude that various harmful biotic
and abiotic factors in water, air and food damage DNA structure in molecules and
thus influence the genome in whole (2,3 ).
CONCLUSION
After completed cytogenetic analyses of 120 cows it can be concluded:
1. The control cow group had good cytogenetic finding 2n=60XX.
2. The experimental group, 70 km away from endangered area, had clean cytogetic
finding 2n=60XX
3. In the experimental group, that was near the area where the harmful influences
were present, three animals with numeric and structural changes (type
aneuploidya and brake on chromosome) were discovered
4. The damage on the chromosome appeared de novo
5. Harmful matters present in the air, water and soil were far above the permitted
values.
6. The investigation is to be continued on the offspring of the experimental group of
cows.
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3. Evans HJ (1982): Cytogenetic studies on industrial populations exposed to
mutagens. Cold Spring Horbor, New York, 324-36
4. Moorhead Ps, Nowel PC, Mellman WJ. Batto[s (1960): Chromosome
preparations from leukocytes cultured from peripheral blood. Exp. cell ress. Ph
20, 613.
5. Gustafsoon A (1991): Mutation environment and evolution. Cole Spring Horbor
Symp. Quant, 16, 263-283
6. Рапорт ИА(1968) Специфичности хемического мутагеноза. Издат Наука
Москва 249