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Health risk assessment of nanomaterials is an emergent field, genotoxicity being an important endpoint to be tested. Since in vivo studies offer many advantages, such as the study of the bioavailability of nanomaterials to sensitive target cells, we propose Drosophila as a useful model for the study of the toxic and genotoxic risks associated with nanoparticle exposure. In this work we have carried out a genotoxic evaluation of silver nanoparticles in Drosophila by using the wing somatic mutation and recombination test. This test is based on the principle that loss of heterozygosis and the corresponding expression of the suitable recessive markers, multiple wing hairs and flare-3, can lead to the formation of mutant clones in larval cells, which are expressed as mutant spots on the wings of adult flies. Silver nanoparticles were supplied to third instar larvae at concentrations ranging from 0.1-10 mM. The results showed that small but significant increases in the frequency of total spots were observed, thus indicating that silver nanoparticles were able to induce genotoxic activity in the wing spot assay of D. melanogaster, mainly via the induction of somatic recombination. These positive results obtained with silver nanoparticles contrast with the negative findings obtained when silver nitrate was tested.

This study presents the first application of an in vivo alkaline comet assay using haemocytes of Drosophila melanogaster larvae. These cells, which play a role similar to that of mammalian blood, can be easily obtained and represent an overall exposure of the treated larvae. To validate the assay, we evaluated the response of these cells to three well-known mutagenic agents: ethyl methanesulfonate (EMS), potassium dichromate (PD), and gamma radiation (γ-irradiation). Third-instar Drosophila larvae were exposed to different concentrations of EMS (1, 2, and 4 mM) and PD (0.5, 1, and 2.5 mM) and to different doses of γ-irradiation (2, 4, and 8 Gγ). Subsequently, haemolymph was extracted from the larvae, and haemocytes were isolated by centrifugation and used in the comet assay. Haemocytes exhibited a significant dose-related increase in DNA damage, indicating that these cells are clearly sensitive to the treatments. These results suggest that the proposed in vivo comet test, using larvae haemocytes of D. melanogaster, may be a useful in vivo assay for genotoxicity assessment., (Published 2010 Wiley-Liss, Inc.)

Few studies on the genotoxicity of mercury compounds have been carried out in Drosophila melanogaster, most of them focused in the effects on germinal cells, whereas studies in somatic cells are scarce. In the present study we have analyzed for the first time the genotoxic activity of mercury (II) chloride (MC) and methyl mercury (II) chloride (MMC) in the in vivo wing somatic mutation and recombination test in Drosophila, also known as the wing spot assay. This test is based on the principle that loss of heterozygosis and the corresponding expression of the suitable recessive markers, multiple wing hairs (mwh) and flare-3 (flr(3)), can lead to the formation of mutant clones in larval cells, which are then expressed as spots on the wings of adult flies. The mercury compounds were supplied to third instar larvae (72+/-2h old) at concentrations ranging from 1 to 50 microM for mercury chloride (MC) and from 0.5 to 5 microM for methyl mercury chloride (MMC). Both mercury compounds showed high toxicity; however, MMC was more toxic than MC. The results showed that none of the three categories of mutant spots recorded (small, large, and twin) increased significantly by the treatments, independently of the dose supplied, indicating that the mercury compounds tested exhibit a lack of genotoxic activity in the wing spot assay of D. melanogaster. These results contribute to increase the genotoxicity database on the in vivo evaluation of mercury compounds in Drosophila.

Inorganic arsenic is nongenotoxic in the Drosophila melanogaster wing somatic mutation and recombination test (SMART). Recent evidence in mammalian systems indicates that methylated metabolites of arsenic are more genotoxic than inorganic arsenic. Thus, we hypothesized that inorganic arsenic is nongenotoxic in Drosophila because they are unable to biotransform arsenic to methylated forms. In the present study, we fed trivalent and pentavalent inorganic arsenic to Drosophila larvae and adults and measured the production of methylated derivatives. No biomethylated arsenic species were found in the organisms or in the growth medium, which suggests that Drosophila are unable to biomethylate inorganic arsenic. Exposure of Drosophila to the methylated arsenic derivative dimethylarsinic acid (DMA(V)) resulted in incorporation of this organoarsenic compound without demethylation. In addition, we used the SMART wing spot assay, which measures loss of heterozygosity (LOH) resulting from gene mutation, chromosomal rearrangement, chromosome breakage, and chromosome loss, to evaluate the genotoxicity of DMA. DMA by itself induced significant increases in the frequency of total spots, small spots, and large single spots. These results are consistent with the important role of arsenic biomethylation as a determinant of the genotoxicity of arsenic compounds. The absence of biomethylation in Drosophila could explain the lack of genotoxicity for inorganic arsenic and the genotoxicity of methylated arsenic species in the SMART wing spot assay., ((c) 2005 Wiley-Liss, Inc.)

In the present study, the herbicides bentazone, molinate, thiobencarb and trifluralin were evaluated for mutagenic and recombinagenic effects using the wing spot test of Drosophila melanogaster (somatic mutation and recombination test, SMART). Both standard (ST) and high-bioactivation (HB) fly crosses were used, the latter cross is characterised by a high sensitivity to promutagens and procarcinogens. Three-day-old larvae, transheterozygous for the multiple wing hairs (mwh, 3-0.3) and flare-3 (flr(3), 3-38.8) genes, were chronically fed with six different concentrations of each herbicide. Feeding ended with pupation of the surviving larvae and the genetic changes induced in somatic cells of the wing's imaginal discs lead to the formation of mutant clones on the wing blade. Point mutation, chromosome breakage and mitotic recombination produce single spots; while twin spots are produced only by mitotic recombination. Bentazone, usually considered as a non-mutagen, gave positive results in the wing spot test with the high-bioactivation cross. Molinate, about which information on mutagenic effects is inconclusive, gave positive responses in both the standard and the high-bioactivation crosses, while the other thiocarbamate, thiobencarb, gave positive results only in the standard cross and at the highest concentration tested (10 mM). Finally, trifluralin, one of the most widely studied herbicides for genotoxic effects, gave positive results in the wing spot test with both crosses. Apart from the interest of the results found in the genotoxic evaluation of the four selected herbicides, our results also contribute to extend the existing database on the Drosophila wing spot test, and corroborate the utility of the use of high-bioactivation strains for the genotoxic evaluation of xenobiotics.

The genotoxic activity of cadmium chloride (CC) has been evaluated in the somatic mutation and recombination test (SMART) in Drosophila melanogaster. In addition, its possible modulating effect on the genotoxicity of two known mutagenic agents, potassium dichromate (PDC) and ethyl methanesulfonate (EMS), was investigated. Three different types of combined treatments of CC with the two genotoxins were performed: pretreatment, cotreatment, and posttreatment. The SMART assay is based on the principle that loss of heterozygosity for the recessive markers, multiple wing hairs (mwh) and flare-3 (flr(3)), leads to the formation of mutant clones in the imaginal disks of larvae, which are expressed as mutant spots on the wings of adult flies. Thus, after adult emergence, the wings of the adult flies were scored for the presence of single and/or twin spots. Our results show that CC alone was not effective in increasing the frequency of any of the three categories of spots (small, large, and twin). In the cotreatment experiments, CC increased the genotoxicity of PDC but it decreased the genotoxicity of EMS. No effects of CC were observed in the pretreatment or posttreatment experiments; however, only low concentrations of CC, PDC, and EMS were tested in the pretreatment assays due to the high toxicity of the treatment. Although our results with PDC are consistent with the hypothesis that cadmium can interfere with repair mechanisms, the EMS data suggest that other modulating mechanisms are also involved in the genotoxicity of this metal., (Copyright 2004 Wiley-Liss, Inc.)

The ability of ascorbic acid (Vitamin C) to modulate the genotoxic action of several mutagens was investigated in the wing spot test of Drosophila melanogaster. In this assay, 3-day-old transheterozygous larvae for the multiple wing hairs (mwh, 3-0.3) and flare (flr, 3-38.8) genes were treated with three reference mutagenic compounds, namely cobalt chloride (CoCl2), 4-nitroquinoline 1-oxide (4-NQO) and potassium dichromate (K2Cr2O7). The results obtained show that the three reference mutagens tested were clearly genotoxic in the Drosophila wing somatic mutation and recombination test (SMART). None of the three concentrations tested of ascorbic acid (25, 75 and 250mM) induced significant increases in the frequency of the mutant clones recorded. When co-treatment experiments with ascorbic acid were carried out, different results were found. Thus, ascorbic acid was effective in reducing the genotoxicity of K2Cr2O7 virtually to the control level; on the contrary, it did not show any antigenotoxic effect on the genotoxicity of 4-NQO. Finally, co-treatments with CoCl2 and ascorbic acid show a significant increase in the frequency of mutant clones over the values obtained with CoCl2 alone.

The wing spot test in Drosophila melanogaster was used to investigate the genotoxicity of arsenic and its effects on the action of two clearly genotoxic agents: potassium dichromate (PDC) and ethyl methanesulfonate (EMS). This assay is based on the principle that the loss of heterozygosity of the suitable recessive markers multiple wing hairs (mwh) and flare-3 (flr(3)) can lead to the formation of mutant clones of larval cells, which are then expressed as spots on the wings of adult flies. These spots can be attributed to different genotoxic events: either mitotic recombination or mutation (deletion, point mutation, and specific types of translocation). Pretreatments and chronic cotreatments were comparatively used for combined treatments. From the results obtained it is evident that sodium arsenite (SA) does not increase the frequency of any of the three categories of spots recorded (small, large, and twin spots) at the concentrations tested. The effects of SA in combination with PDC, in both cotreatments and pretreatments, indicate that SA almost suppressed the clones induced by PDC. Nevertheless, no effects of arsenic were observed with respect to the pre- and cotreatments with EMS. Thus, SA does not modify the frequencies of mutant clones induced by EMS., (Copyright 2002 Wiley-Liss, Inc.)

Genotoxic and carcinogenic effects have been attributed to disinfection by-products (DBP5), but few studies on the genotoxicity of hydroxyfuranones (HFs) exist. This limited information gives special relevance to obtaining new data on their potential genotoxic activity, mainly by in vivo test approaches. In this study we present the results obtained with two HIPs, namely mucochloric acid (MCA) and mucobromic acid (MBA), in the in vivo wing somatic mutation and recombination test in Drosophila, which detects both somatic mutation and recombination, two mechanisms of especial relevance in cancer processes. The results showed that none of the three categories of mutant spots recorded (small, large, and twin) increased significantly by the treatments, independently of the concentrations tested. This indicate that the two I-IFs evaluated do not exhibit genotoxic activity in the wing spot assay of D. melanogaster. These in vivo results contribute to increase the scarce genotoxicity database on the DBPs. [ABSTRACT FROM AUTHOR]

Purpose: The aim of this study was to assess the adaptive response induced by low doses of ultraviolet-B (UVB, 290-320 nm) radiation in the Drosophila wing spot test. Materials and methods: The adaptive response of Drosophila larvae to UVB light was studied by using a somatic mutation and recombination test (SMART). The SMART system used was the wing spot test, which uses morphological markers of the wing blade. This in vivo test has shown to be very useful to study the induction of genetic damage in somatic cells, measuring loss of heterozygosity (LOH) resulting from gene mutation, mitotic recombination, chromosomal rearrangements or chromosome loss. Results: To determine the induction of adaptive response, two-day-old Drosophila larvae were first irritated with an adaptive dose (58.3 J/m2), followed by different challenge doses (178, 224, 288, 338, and 386 J/m2). When the results obtained in the different challenge doses were compared with those obtained following the application of adaptive plus challenge doses, significant decreases (74.7-80.8%) in a first experiment, and (65.6-78.4%) in a second experiment, were observed in the frequency of mutant spots on the wing blades. Conclusions: Our results show that in Drosophila the adaptive response can be stimulated in vivo by UVB exposure. [ABSTRACT FROM AUTHOR]

Few studies on the genotoxicity of halonitromethanes (HNMs) have been done. This limited information on their potential genotoxic risk gives special relevance to the collection of new data on their potential genotoxic activity. In the present study we have analyzed the genotoxicity of two HNMs namely bromonitromethane (BNM) and trichloronitromethane (TCNM) in the in vivo wing somatic mutation and recombination test in Drosophila, also known as the wing-spot assay. This test is based on the principle that loss of heterozygosis and the corresponding expression of the suitable recessive markers, multiple wing hairs (mwh) and flare-3 (flr3 ), can lead to the formation of mutant clones in larval cells, which are then expressed as spots on the wings of adult flies. BNM and TCNM were supplied to third instar larvae (72±4h-old) at concentrations ranging from 0.1 to 2mM. The results showed that none of the three categories of mutant spots recorded (small, large, and twin) increased significantly by the treatments, independently of the dose supplied, indicating that the selected HNMs exhibit a lack of genotoxic activity in the wing-spot assay of Drosophila melanogaster. These results contribute to increase the genotoxicity database on the HNMs. [Copyright &y& Elsevier]