1067-4136/04/3506- © 2004
Russian Journal of Ecology, Vol. 35, No. 6, 2004, pp. 426–430. From Ekologiya, No. 6, 2004, pp. 473–476.
Original English Text Copyright © 2004 by Milo evi -Djordjevi , Gruji i , Arsenijevi , Marinkovi .s
Since 1985, the cytokinesis-block micronucleus
(CB MN) assay in human peripheral blood lympho-
cytes has been accepted by many laboratories all over
the world as an optimal method for assessing chromo-
some damage (Fenech and Morley, 1985).
The basic advantage of this test is that it allows an
analysis of a great number of lymphoblasts per sample,
which is important for obtaining a more realistic picture
of cytogenetic damage (Bonassi
, 1995; Fenech,
1997; Suralles and Natarajan, 1997).
The fact is that, in 1999, the Zastava factory in Kra-
gujevac was bombed several times, and the bombing
caused heavy material destruction and endangered the
human environment. In the ﬁres caused by the bomb-
ing, 95000 kg of different materials and commodities
burned, including 2200 kg of a sealant and 800 kg of
pyralen, which belongs to polychlorinated biphenyls
(PCBs). Pyralen oil poured out of two damaged trans-
formers (1428 l per transformer) and partly remained in
the holes for liquid waste decantation. The remainder,
after the ﬁres, partly spread over the industrial zone and
partly ﬂowed into rivers, lakes, etc.
These harmful materials evaporate slowly and dis-
solve in lipids. They enter the human body in several
ways: through the food chain, by inhalation/breathing,
by ingestion, or through the skin and they accumulate
in adipose tissues. PCBs have acute toxicity and sec-
ondary effects connected with chronic exposure
The main aim of this biomonitoring study was ade-
quate evaluation of cytogenetic damage in samples of
newborns from Kragujevac by analyzing spontaneous
MN frequency in peripheral blood lymphocytes.
MATERIALS AND METHODS
The analyzed sample comprised a total of 61 pheno-
typic healthy newborns delivered at the Department of
Obstetrics and Gynecology in Kragujevac in the period
from 1998 to 2002. The control value of MN frequency
was obtained by analyzing peripheral blood lympho-
cyte cultures derived from 25 newborns before the
bombing; after the bombing, a sample of 36 newborns
was analyzed in the same way.
For our cytogenetic study, the classical method for
culturing human peripheral blood lymphocytes was
used with some modiﬁcations that are necessary for
applying the CB MN test (Edwards, 1962; Frland,
1962; Fenech, 1997).
The Frequency of Micronuclei among Newborns
from Kragujevac, Central Serbia, after NATO Bombing
in the Spring of 1999
O. Milo evi -Djordjevi
, D. Gruji i
, S. Arsenijevi
, and D. Marinkovi
Faculty of Science, Kragujevac, Serbia and Montenegro
Clinic of Gynecology, Kragujevac, Serbia and Montenegro
Faculty of Biology, Belgrade, Serbia and Montenegro
Received September 29, 2003
—Kragujevac, as an important industrial and economic center of the region, is now placed on the top
of the list of environmentally endangered cities in our country. The aim of this biomonitoring study was to eval-
uate cytogenetic damage in a sample of newborns from Kragujevac after contamination of the environment
entailed by the intensive bombing of the industrial zone of this city in the spring of 1999. The frequency of
micronuclei (MN) in peripheral blood lymphocytes in a total of 61 phenotypically healthy newborns was ana-
lyzed before and after the NATO bombing. Analysis of micronuclei has been performed using the cytokinesis-
block technique (CB MN test). Average MN frequency in lymphocytes of newborns before the bombing (
25) was 5.77
0.85/1000 analyzed cells. After the bombing (
= 36), the average frequency of MN increased
by a factor of 1.4 (8.11
0.85), compared to the control frequency before the bombing. Statistical difference
< 0.05) was established by Student’s
-test. Our data suggest that such changes in genetic material were a
direct consequence of contamination of the living environment.
: micronucleus test, environment, technogenic pollution, human population, Serbia.
This article was submitted by the authors in English.