Sandra Morelli

Cytogenetic Considerations on the Genus Astyanax (Pisces, Characidae). I. Karyotypic Variability

SUMMARYA study was carried out on mitotic and meiotic chromosomes of four species of the genus Astyanax (Pisces, Characidae) from three different water basins in the State of Sao Paulo, Brazil. The following karyotypic characteristics were observed: A. bimaculatus, 2n = 50 (n = 25, FN = 86–88); A. fasciatus, 2n = 46 (n = 23, FN = 90); « A. fasciatus » from the Juquia river, 2n = 48 (n = 24, FN = 94); A. schubarti, 2n = 36 (n = 18, FN = 70) and A. scabripinnis paranae, 2n = 50 (n = 25, FN = 90–92). The diversity in the number and morphology of the chromosomes suggests a model of non-conservative karyotypic evolution in this genus, with chromosome rearrangements of the centric fusion/fission type and/or inversion playing an important role in these alterations. On the basis of its characteristics, A. schubarti has the most differentiated karyotype and, in view of the low diploid number and small number of subtelocentric-acrocentric chromosomes, is probably of most recent origin. The two karyotypes encountere...

In situ biomonitoring of the genotoxic effects of vehicular pollution in Uberlândia, Brazil, using a Tradescantia micronucleus assay.

The growing number of cars in large cities is directly linked to changes in the chemical composition of urban air, which has increasingly high concentrations of potentially genotoxic chemicals. Therefore, discovering and monitoring the risks associated with exposure to atmospheric pollutants is indispensible for preventing environmental and health problems. Because of the lack of reliable data regarding the air quality in the city of Uberlândia, the present study sought to test whether the genotoxic risks in areas with different levels of vehicular traffic can be measured using the Tradescantia micronucleus assay (Trad-MN). Therefore, more than twenty inflorescences were exposed to locations with different amounts of vehicular traffic twice per year from the winter of 2006 to the summer of 2011. The inflorescences were then analysed to determine the micronucleus (MN) frequency. In addition, we sought to determine the influence of factors linked to city climate on the MN frequencies obtained at each monitored location. Our results show that, although low relative humidity positively influenced MN formation in Tradescantia pallida tetrads, the major determining factor for clastogenic events was the level of vehicular traffic at the locations monitored over the five-year study.

Cytogenetic Considerations on the Genus Astyanax (Pisces, Characidae). II. Occurrence of Natural Triploidy

SUMMARYAstyanax schubarti is characterized by a diploid number of 36 chromosomes. In the present paper we report on the occurrence of a specimen with 54 chromosomes, a probable case of natural triploidy as shown not only by the chromosome number but also by the good morphological correspondence between the chromosomes of the 3x and 2x individuals. The fertilization of an unreduced oocyte is postulated as the probable cause of this triploidy.

In vitro and in vivo antitumor effect of trachylobane-360, a diterpene from Xylopia langsdorffiana.

Trachylobane-360 (ent-7α-acetoxytrachyloban-18-oic acid) was isolated from Xylopia langsdorffiana. Studies have shown that it has weak cytotoxic activity against tumor and non-tumor cells. This study investigated the in vitro and in vivo antitumor effects of trachylobane-360, as well as its cytotoxicity in mouse erythrocytes. In order to evaluate the in vivo toxicological aspects related to trachylobane-360 administration, hematological, biochemical and histopathological analyses of the treated animals were performed. The compound exhibited a concentration-dependent effect in inducing hemolysis with HC50 of 273.6 µM, and a moderate in vitro concentration-dependent inhibitory effect on the proliferation of sarcoma 180 cells with IC50 values of 150.8 µM and 150.4 µM, evaluated by the trypan blue exclusion test and MTT reduction assay, respectively. The in vivo inhibition rates of sarcoma 180 tumor development were 45.60, 71.99 and 80.06% at doses of 12.5 and 25 mg/kg of trachylobane-360 and 25 mg/kg of 5-FU, respectively. Biochemical parameters were not altered. Leukopenia was observed after 5-FU treatment, but this effect was not seen with trachylobane-360 treatment. The histopathological analysis of liver and kidney showed that both organs were mildly affected by trachylobane-360 treatment. Trachylobane-360 showed no immunosuppressive effect. In conclusion, these data reinforce the anticancer potential of this natural diterpene.

The impact of water pollution on fish species in southeast region of Goiás, Brazil

ABSTRACT The rivers from the region of Catalão, Southeast Goiás State, Brazil, are exposed to intense anthropogenic influences including agricultural activities, industry, and urban waste. The aim of this study was to determine the severity of water pollution by conducting an experiment involving in situ biomonitoring of water constituents on genotoxicity in fish inhabiting these sites. The genotoxicity of three sites of the region were analyzed utilizing the micronucleus (MN) test. It was of interest to determine whether there were differences between sampling sites such as urban perimeter, agriculture, and fertilizer industry in control, and monitored species including Astyanax fasciatus, Astyanax altiparanae, and Characidium fasciatum. Data demonstrated that the species at sites 1, 2, and 3 exhibited a marked increase in frequency of MN compared to fish from site 4. Significant elevation in frequency of MN occurred in erythrocytes of A. fasciatus and A. altiparanae at sites 1 and 2. At site 3 higher frequencies of MN were observed in C. fasciatum. MN induction in C. fasciatum was correlated with chromium levels in water and sediment, while A. fasciatum and A. altiparanae showed an association with zinc in water and sediment. Data suggest that benthic and nektonic fish species display different sensitivities in relation to anthropogenic contaminant influences.

Monitoring Genotoxicity Potential in the Mumbuca Stream, Minas Gerais, Brazil

Rivers are sites for water catchment to supply metropolitan areas but also serve as receptors for discharge of urban sewage, wastewater, and agri-industrial effluents. Bioindicators or sentinel organisms are widely used as markers of pollution in various environments. The objective of this study was to evaluate the genotoxic potential and consequent quality of the water from the Mumbuca stream, which supplies the city of Monte Carmelo, located in the Minas Triangle region, Minas Gerais, Brazil. This was achieved using two variable response bioindicators (Rhamdia quelen and Geophagus brasiliensis), the micronucleus (MN) test, and determining the presence of metals by flame atomic absorption spectrometry. Results showed that site 3 water (region of residential flow and intense industrial pottery activity) presented a greater possibility for induction of genotoxic activity, as evidenced by the increase in the MN frequency in Rhamdia quelen and Geophagus brasiliensis in comparison with the reference-site water. The water of the Mumbuca stream was influenced by genotoxic agents, especially lead and chromium, assessed by the rise in MN rate. Data suggested that discharge of industrial effluents in a specific stretch of the stream interfered with biota functions.

Cytogenetic analysis in the neotropical fish Astyanax goyacensis Eigenmann, 1908 (Characidae, incertae sedis): karyotype description and occurrence of B microchromosomes

BackgroundB chromosomes, also known as supernumerary or accessory chromosomes, are additional chromosomes over the standard complement found in various groups of plants and animals. We investigated the presence of, and characterized, supernumerary microchromosomes in Astyanax goyacensis using classical and molecular cytogenetic methods.FindingsThree specimens possessed 2n = 50 chromosomes (8m + 26sm + 8st + 8a), and two specimens contained 1 to 9 additional B microchromosomes varying intra- and inter-individually. Chromosome painting with a B chromosome-specific probe yielded signals for several B microchromosomes, with one exhibiting no markings. Acrocentric chromosomes of the standard complement were also painted. Fluorescence in situ hybridization (FISH) using ribosomal probes located two chromosome pairs carrying 18S rDNA marked on the short arm, and one pair carrying 5S rDNA with pericentromeric markings. One chromosome was observed in synteny with 18S cistrons.ConclusionThese data contribute to knowledge of the karyotype evolution, the origin of B chromosomes, and to an understanding of the functionality of rDNA.

Evolutionary cytogenetics of the Hoplias lacerdae, Miranda Ribeiro, 1908 group: a particular pathway concerning the other Erythrinidae fish

The taxonomy/systematics of the Erythrinidae fish is still imprecise, with several doubts on their relationships. Karyotypes and chromosomal characteristics of some species of the Hoplias lacerdae group (Erythrinidae), from different Brazilian hydrographic basins and pisciculture stations, were analyzed in the present study, using conventional Giemsa staining, C-banding, silver staining, Mithramycin and Distamycin/DAPI fluorochromes, and fluorescent in situ hybridization (FISH). A diploid chromosome number of 2n = 50 and karyotypes composed of meta- and submetacentric chromosomes without sex-related differences were found. Only one active NOR (Nucleolar Organizer Region) site was found, which was identified by silver staining (Ag-NOR) and FISH, located on the chromosome pair 11, although additional 45S rDNA sites were also mapped on other chromosome pairs only by FISH. The Ag-NOR of the chromosome pair 11 was found to be GC-rich, appearing positive after Mithramycin staining. Mithramycin-positive/DAPI-negative sites were also observed in the centromeric/pericentomeric regions of the chromosome pairs 4, 6, 15, and 19, which have also affinity to silver nitrate. However, these four sites were not detected by FISH with the rDNA probe, indicating to be only argentophilic GC-rich heterochromatic regions. Chromosome data show that the karyotype evolution in Hoplias lacerdae group is relatively conserved and follows a particular pathway concerning the other Erythrinidae fishes, such as Hoplias malabaricus, Hoplerythrinus unitaeniatus, and Erythrinus erythrinus, in which polytypic karyotypes are found. Thus, the H. lacerdae group shows chromosome features that are not closely related to those of the congeneric H. malabaricus group. These finds, together with genetic and morphologic data, are important tools to be considered in a major revision of the Erythrinidae family, as well as for conservation programs.

Biomonitoring air quality during and after a public transportation strike in the center of Uberlândia, Minas Gerais, Brazil by Tradescantia micronucleus bioassay.

The aim of this study was to address the lack of information concerning the air quality in the city of Uberlândia, Minas Gerais, Brazil. In this study, we conducted an unprecedented experiment involving the in situ biomonitoring of air genotoxicity in the city center during and after a public transportation strike using the Tradescantia micronucleus test. The frequency of micronuclei was significantly higher in the city center compared with the reference site (Mann–Whitney test, p < 0.05), with the highest MN levels being observed during public transport stoppage (Kruskal–Wallis, Dunn p < 0.01). In addition, the multiple linear regression analyses revealed that the low circulation of buses during public transport stoppage and the increase in the concentration of particulate matter from the increased flow of vehicles in the city center during the strike positively influenced the MN frequency. The climatic factors did not change during the biomonitoring period, reflecting the fact that climatic factors did not influence the MN frequency.

Toxicological assessment of spinosad: Implications for integrated control of Aedes aegypti using larvicides and larvivorous fish

ABSTRACT Integration of larvivorous fish and biolarvicides at low concentrations to control of mosquito larvae in field situations may result in a safer and more effective tool. However, the usefulness of integrated approach depends upon survival and ecological fitness of fish employed. Thus, the aim of this study was to examine the genotoxic effects of combining different sublethal concentrations of spinosad, a naturally occurring neurotoxic insecticide, with male adult poecilid larvivorous guppy (Poecilia reticulata) and platy (Xiphophorus maculatus) fish on Aedes larvae mosquitos. Both fish species have been used for biological control of Aedes larvae in Brazil. Sublethal spinosad exposures were predetermined based on CL50-96hr. Nuclear abnormalities (NA) and micronucleus (MN) frequency in gill cells were measured after 14 d of exposure. Behavioral changes were monitored over 96 h. Although genotoxic effects were not markedly different from control, behavioral changes evaluated based upon the no-observable-effect concentration (NOEC) and lowest-observable-effect concentration (LOEC). Adverse effects were noted at concentrations of 12.6 mg/L (NOEC) and 25.3 mg/L (LOEC) spinosad. Therefore, these insecticide concentrations may be considered as being safe to these fish species and have important implications for integrated approach to control Aedes larvae using natural larvicides and larvivorous fish.