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Sistema de Información Científica
Red de Revistas Científicas de América Latina y el Caribe, España y Portugal
Rev. Int. Contam. Ambient. 5, 5944, 1989.
THE HERBICIDES DALAPON
AND
DIURON TESTED FOR
GENOTOXICITY IN
Drosophila melanogaster
Rosario RODR~GUEZ-ARNAIZ', Patricia RAMOS MORALES', Juan Carlos
GAYTÁN
OYARZUN', Dora Lydia RODR~GUEZ
zÚÑIGA'
and Stanley
ZIMMERING~
'
Laboratorio de Genttica, Facultad de Ciencias, UNAM, Ciudad Universitaria, Cqoacán 04510 D.F.,
México.
Divicion of Biology and Medicine,
Brown
University, Providencc, Rhode lsland 02912,
USA.
(Recibido septiembre 1989, aceptado diciembre
1989)
The herbicides dalapon and diuron, used in large quantities
in
Mexico, were
evaluated for genotoxicity in
Drosophila melanogasfer.
Tests for the induction of
sex-linked recessive lethals were carried out with both compounds. With diuron
only, a test for sex chromosome loss using a repair deficient system was carried out
in addition. Under the conditions of experiments, dalapon proved negative and
diuron positive
in
the recessive lethal test. The results obtained for the sex
chromosome loss test are largely negative. There is no indication of a true genotoxic
effect of diuron in this assay.
RESUMEN
Se evaluaron los efectos genotóxicos de los herbicidas dalapón y diurón, amplia-
mente usados en México. Para ambos compuestos se analizó la inducción de
mutaciones letales recesivas ligadas al sexo. La prueba de pérdida de cromosomas
sexuales, utilizando un sistema deficiente en reparación, se corrió solamente para
el diurón. Bajo las condiciones experimentales el dalapón ~esultó
negativo y el
diurón positivo en el estudio de letales recesivos. Los resultados obtenidos en la
prueba que mide la pCrdida de los cromosomas sexuales son negativos. No hay
indicación de efectos verdaderamente genotóxicos del diurón en este ensayo.
INTRODUCTION
Among the environmental chemicals with possible hazards to man, the pesticides play
an important role since residucs of these substancies may remain in food and be ingested
by humans (Duggan and Weatherwax 1%7). With the dispersa1 of hundreds of millions
of kilograms of pesticides on hundred of millions of hectares of land around the world,
prudence requires that the mutagenic properties of these chemicals be determined and
that a variety of bioassay systems be used in the process (Plewa et al. 1984).
Dalapon (2, 2 dichloropropionic acid
sodium
salt), a permeation-spreading her-
bicide,
is
employed
in
orchards and fields (Tsukioka and Shimizu 1985) to erradicate
deep-rooted perennial grasses (Weeraratna 1980). The herbicide
is
soluble in water at
ambient temperature and neither photodecomposes nor e-yaporates; nevertheless, it
is
biodegradable by dehalogenation producing inactive pyruvate (Matswnara 1973, Audus
1976, Barbera 1976). About 228,821 tons of dalapon were irnported into Mexico during
the 1985-1986 period (Productora Nacional de Semillas, personal comrnunication).
Diuron (3-(3, 4dichlorophenyl)-, 1-dimethylurea), a ureic herbicide, attacks roots of
growing plants (Majka and Levis 1977). Herbicidai uses of diuron are numerous on both
crops and noncrop land; it is used primarily to control annual grass and broadleafweeds
before emergente in alfalfa, artichokes, asparagus, barley, blueberries, corn, cotton,
grapes, pineapple, sorghum, sugarcane and nut tree crops(Wingman et al. 1975). D'
luron
has low water solubility and
is
extremely persistent in soils although it can ultimately be
degraded by microbial action (Attaway et
al.
1982). It acts by strongiy reducing the
photosynthetic activity of plants by inhibition of the HiU reaction in the chloroplast
(Brown 1978). The production of diuron in Mexico was about 320 tons (Productora
Nacional de Semillas, personal communication).
The present paper deals with the genotoxic effects of dalapon and diuron in the
sex-linked recessive lethal test and of diuron additionally in a sex chromosome loss test
in
Dtosophila melanogaster.
MATERIAL
AND
METHODS
In the test to screen for sex-lied recessive lethals (SLRLT), Oregon-R wild
type
males and females carrying a balancer X-chromosome, Basc, with the genotype In(1)
scS1Lsc8R
+
S,
SC~~SC*W~B
were used. For the sexchrornosome loss test (SCLT) the males
carried a ring X-chromosome
marked with the recessive mutants y
f
bb- and a
1
2,
special Y chromosome
Y y
,
bearing the dominant marker BS on the Ion arm of
the Y chromosome,
Y:
and the dominant alleles ofy and bb,
and bbg on the
short arm,
Y'
(Brosseau 1958). Males were mated with repair-deficient females of the
constitution y ac sc wa mei-9a. It is known that mei41a is a meiotic mutant as well as a
rcpair-deíicient niutant that strongly affects excision repair (Baker and Carpenter 1972,
Boyd et al. 1976). It sensitizes the sex chromosome loss test to the genetic effects of many
chemical mutagens (Zimmering 1983). For an explanation
of
genetic symbols see
Lindsley and Grell (1968).
DALAPON AND DIURON IEXED FOR GENOTOXlCrrY
Test for recessive lellrals:
Treatcd and control males were mated individually with
thrcc virgin femalcs to produce thrce broods: Brd
A
(two days), Brood B (three days)
and Brd C (two days); thus primarily postmeiotic and late meiotic germ cells were
tested (Chandley and Bateman 1%2). Fi females were mated individudy to sibs and F2
progeny were scored for the presence or absence of wild type males.
A
lethal mutation
is
taken lo havc occurred if no wild type males were recovered in
Ft
among at least 16
Bascl
+
females or if wild type males were less than
5
%
of the total
Bmcl+
females.
Suspected lcthals were retestrd in F3.
Test /or clirmtosorne loss:
Treated and control males were mass mated in bottles
with virgin females and transfcrred so as to produce three broods of 2-3-2 days.
Rcylar
Fi
femalcs are phenotypicallyy aad
Ft
males
wa
B'.
Complete 106s (CL) of
X or Y generates a phenotypically y
ac
wa
male, while partial loss (PL) of the Y
chromosome in thc case of loss of thc y+ marker from the Y-chromosome produces ay
oc wa E'
male and a
wa
male in the case of loss of the
B'
rnarker.
Chemicals ond solvents:
Dalapon (trade names Basfpon, Dowpon) was obtained
from BASF (Mcxico City), diuron (trade names Karmex, Telvar, Urox D) from Du Pont
(Mcxico City). Thc solvent for dalapon was a solution of 5
%
sucrose, and for diuron a
solution of 1.5
%
DMSO.
Treatnrent:
For the SLRLT experiments, adult males 0-48 hours old, were injected
with 0.2-2.3~1
of fresh solution. For the SCLT experiments adult males wcre fed during
24 hours on a filtcr disc placed at the bottom of vials saturated with 0.5-0.7 ml of the
solution for 24 hours and then transferred to vials with fresh solutions for an additional
24 hours.
Statistical analysis:
The cumulative Poisson (Owcn 1962) was crnployed to identify
clusters. After corrcction for clusters (if any) the data were analyzcd with the Kasten-
baum-Bowrnan test (1970) to determine the significance of the difference betwcen
control and treated values.
RESULTS AND DISCUSSION
Results of the
Basc
test for sex-linked reccssive lethal rnutations after treatment with the
herbicidcs by injcction of adult Oregon-R males are shown in Table
1
and 11. Tablc
1
shows thc results obtained after trcatmcnt with dalapon. It can bc secn from this table
that there is no significant diíTercnce bctween control and trcatcd values at 5
%
lcvel in
any of the broods at any of the concentrations. Thcse rcsults are in agreemcnt with the
negative findings reportcd by Siebert and Lempcrle (1974) for mitotic conversion test
in
Sacclrarornyces cereviseae
and wit h t hosc by Moriya
ci
al.(1983),
who found no evidcncc
that dalapon significantly increases the frequcncy' of rcvcrsions in the Salmoncl-
TABLE 1. RESULTS OF THE SEX-LINKED RECESSIVE LETHAL TEST IN
Drosophila
melanogaster
AFTER INJECTION OF THE HERBICIDE DALAPON INTO
ADULT OREGON-R WILD TYPE MALES. THE SYMBOLS
#
l/# tests SIG-
NIFY THE NUMBER OF LETHALS IDENTIFIED IN A TEST SAMPLE OF
XCHROMOSOMES. THE DATA REPRESENT THE SUM OF TWO EXPERI-
MENTS
Concentration
Brood A
Brood B
Brood C
Broods A +B+C
(PP~)
(0-2 &YS)
%
(3-5 &y#)
%
(6-7&~~)
%
(O-7&ys)
%
#
I/#
tests
#
1/#
tests
#
1/#
tests
#
I/#
tests
Control
5%
sucrose
011175
011616
011233
014024
TABLE 11. RESULTS OF THE SEX-LINKED RECESSIVE LETHAL TEST IN
Drosophi-
la
melanogaster
AFTER INJECTION OF THE HERBICIDE DIURON INTO
ADULT ORECON-R WILD TYPE MALES. THE SYMBOLS *ll/+tests
SIGNIFY THE NUMBER OF LETHALS IDENTIFIED IN A TEST SAMPLE
OF XCHROMOSOMES. THE DATA REPRESENT THE SUM OF THREE
EXPERIMENTS
Concentrarion Brd A
Brood B
Brood C
Broodr A +B+C
(PP~)
(0-2 &ys)
(3-5 Lys)
(6- 7 &YS)
(O- 7 d a y s )
#
I/#
tests
%
#
i/#
tests
%
#
ll#
tests
%
#
ll#
tests
%
Control
DMSO 1.5%
111662
0.06 011324
211289
0.15 312475
0.07
*p=0.05
'(t.
la/microsome test. The frcquencies of lcthals recovered after diuron treatment are given
in Table
11.
This herbicide induced significant increases in the frequency of recessive
lethals in Broods
A
and B at a concentration of
40
ppm. It may be recalled that Broods
A
and B represent postmeiotic stages, principally mature sperm and late spermatids.
The mean frequency over the three broods is 0.38
%
a value some 5 to 6 times greater
than the control (0.07
%).
Results of experimcnts testing for scx chromosomes loss with
DALAPON
AND
DIURON
TESTED
POR GENOTOXIW
diuron are shown in Table 111. The results obtained are largely negative. It was not
possible to test the ring-X males at
40
pprn
because
of the toxic effects of the chemid
at that concentration.
Briefly, the herbicide dalapon was negative
in
the test for sex-linked recessive lethals
in Drosophiia whereas the herbicide diuron was positive
in
that test and negative
in
the
test for sex chromosome loss using repair-deficient females
mei-98.
TABLE 111. COMPLETE CHROMOSOME LOSS (CL), PARTIAL CHROMOSOME LOSS (PL)
AND MOSAICS FOUND IN THE PROGENY OF DIURON-TREATED MALES
WITH REPAIR-DEFICIENT
mei-9a
FEMALES OF Drosophila
melanogaster.
THE DATA REPRESENT THE SUM OF THREE EXPERIMENTS
Concentration
Brood
Total
Regulnr
Regulnr
Chromosome Loss
Mosaics
(PP~)
F1
females
males
CL
%
PL
%
%
Control
DMSO
1.5%
A
3144
2033
1085
20
0.64
5
B
2163
1141
1008
12
0.55
2
C
1820
1033
770
15
0.82
2
A+B+C
7127
4207
2863
47
0.66
9
ACKNOWLEDGEMENTS
We
thank
Raque1 Ortiz Martelio and Alfredo Delgado Rodríguez for their excellent
technid assistance.
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