Bruno do Amaral Crispim

Genetic instability in plants associated with vehicular traffic and climatic variables

To characterize the effect of vehicular traffic on air quality, the micronuclei of Tradescantia pallida tetrads were counted. Young inflorescences of T. pallida (Rose) D.R. Hunt var. purpurea were collected in 2010, 2011, and 2012, from three sites subjected to different intensities of vehicular traffic. The sites were located in the municipality of Dourados, in the State of Mato Grosso do Sul, Brazil. A standardized methodology was used to analyze the Tradescantia micronuclei, in order to evaluate the mutagenic potential of the local air pollutants. Statistical analyses using the Pearsons linear correlation were employed to determine the relationship between relative humidity and temperature, and the average number of micronuclei. In this study, an increase in the average number of passing vehicles was correlated with an increase in the frequency of micronucleus formation. Climatic factors also influenced micronucleus formation, although vehicular traffic remained the most important factor. Thus, the Tradescantia micronuclei assay may be a useful method of assessing air quality.

Genotype, antifungal susceptibility, and biofilm formation of Trichosporon asahii isolated from the urine of hospitalized patients

In this study, the culture analysis of urine samples from patients hospitalized in the Central-West region of Brazil was performed, and the isolated microorganisms were phylogenetically identified as Trichosporon asahii. Maximum parsimony analysis of the IGS1 sequences revealed three novel genotypes that have not been described. The minimum inhibitory concentrations of the nine isolates identified were in the range of 0.06-1 μg/ml for amphotericin B, 0.25-4 μg/ml for fluconazole, and 0.03-0.06 μg/ml for itraconazole. Approximately 6/9 of the T. asahii isolates could form biofilms on the surface of polystyrene microplates. This study reports that the microorganisms isolated here as T. asahii are agents of nosocomial urinary tract infections. Furthermore, the IGS1 region can be considered a new epidemiological tool for genotyping T. asahii isolates. The least common genotypes reported in this study can be related to local epidemiological trends.

Effects of atmospheric pollutants on somatic and germ cells of Tradescantia pallida (Rose) D.R. HUNT cv. purpurea

Anatomical alterations in leaves and DNA damage in cells caused by the accumulation of atmospheric pollutants can be measured by epidermal leaf analyses and Tradescantia micronuclei assay with early pollen tetrad cells. The present study examined the feasibility of using somatic and germ cells of Tradescantia pallida for biomonitoring purposes in the city of Dourados, state of Mato Grosso do Sul (MS), Brazil. Stomatal, micronucleus and epidermal leaf analyses were performed, using standard methodologies, on plants growing at three locations during six different time periods. Tradescantia micronuclei data were analyzed using SAS 9.2 software package and stomatal data were analyzed using SANEST software. Analyses of stomatal characteristics and micronuclei examination in T. pallida were found to be an efficient tool for monitoring atmospheric pollution. The micronucleus assay suggested that the number of micronuclei in early pollen tetrad cells was related to the intensity of vehicular traffic. Increased number of epidermal cells and stomata and increased stomatal density observed at locations with greater vehicular traffic are likely physiological responses of those plants to the increased gas exchange in highly polluted environments.

Association of the leptin gene with carcass characteristics in Nellore cattle.

Advances in DNA technology have created biotechnological tools that can be used in animal selection and new strategies for increasing herd productivity and quality. The objective of the present work was to associate the genotypes of leptin gene exon 2 polymorphisms with productive traits in Nellore cattle. Blood was collected from Nellore males and PCR-RFLP reactions were performed with the restriction enzymes ClaI and Kpn2I. The gene frequencies resulting from digestion by ClaI were 0.60 and 0.40 for allele A and T, respectively; the genotypic frequencies were AA = 0.20 and AT = 0.80. The gene frequencies from digestion by Kpn2I were 0.81 for allele C and 0.194 for allele T; the genotypic frequencies were CC = 0.62 and CT = 0.38. The populations in both cases were not in Hardy-Weinberg equilibrium (p > 0.05), and the TT genotype was not found. Significant associations were noted between leptin gene exon 2 polymorphisms and five productive traits in Nellore cattle: carcass fat distribution, the intensity of red muscle coloration, pH, marbling, and post-slaughter fat thickness.

Identification of class 1 and 2 integrons from clinical and environmental Salmonella isolates

INTRODUCTION The indiscriminate use of antimicrobials has selected for the emergence of resistant strains. Many mechanisms contribute to the spread of antimicrobial-resistant genes, and integrons play a key role in this process. The aim of this study was to describe the serotypes and resistance profiles, and to characterize the presence of integrons in Salmonella strains isolated from Dourados, Mato Grosso do Sul, Brazil. METHODOLOGY Thirty-six isolates from different sources were used. To evaluate the resistance profiles, the determination of minimum inhibitory concentrations together with polymerase chain reaction were used to screen for the presence of class 1 and class 2 integrons. RESULTS The Infantis serotype of Salmonella was the most frequently isolated serotype. Minimum inhibitory concentrations showed that out of the 36 isolates, 11 (30.5%) were resistant to all the antimicrobials tested. These resistant isolates were separated into three groups: 4 clinical isolates (36.4%), 3 food isolates (36.4%), and 4 water isolates (27.2%). Class 1 integrons occurred in 31 (86.1%) isolates and were found in all 11 resistant isolates (35.5 %) and in 20 (64.5%) of the non-resistant isolates. Class 2 integrons were found in 3 (8.3%) isolates, which were all non-resistant. CONCLUSION The presence of an integron did not necessarily confer resistance. Future studies will seek to identify the mechanism behind integron-mediated antimicrobial resistance.

Genotoxic and mutagenic effects of passive smoking and urban air pollutants in buccal mucosa cells of children enrolled in public school

Abstract Nuclear abnormalities (micronuclei and meta-nuclear changes) have been used as biomarkers to identify cell damages. As children are more vulnerable to the adverse effects of pollution when compared to adults, assessing genetic damage caused by environmental influences is of great interest. As such, the objective was to determine metanuclear (karyolysis, pycnosis, karyorrhexis, binucleated cells, chromosome bridges and micronuclei) in cells from the oral mucosa of children associated with the school environment, gender, exposure to cigarette smoke and vehicular traffic. Analyses of nuclear abnormalities were performed in exfoliated buccal cells of children from two public schools located in Dourados – MS. The data were analyzed through Kruskal–Wallis test considering a significance level of 5% (p < .05). The results showed that children exposed to cigarette smoke presented higher levels of nuclear abnormalities than children who were not usually exposed to this type of mutagenic and genotoxic agent, suggesting that such contaminants are related to clastogenic and aneugenic effects on DNA. Moreover, female children had higher amounts of nuclear abnormalities when compared to male children. With regards to the school environment, the study results indicated statistical differences in of term chromosomal abnormalities for schools A and B. Thus, it was possible to determine that children exposed to cigarette smoke are susceptible to further genetic damage than unexposed children, and female children may be more susceptible to genotoxic and mutagenic agents. This study contributes to the current knowledge on the mutagenic characteristics of human cells, supporting the adoption of preventive Public Health measures.

Depth and Well Type Related to Groundwater Microbiological Contamination

Use of groundwater from private wells in households has increased considerably, owing to a better cost/benefit ratio than that of water provided by local utilities for a fee. However, this water is usually untreated, which makes it a vehicle for diseases. Thus, monitoring this water is necessary to ensure its integrity and quality. We aimed to evaluate the physical, chemical, and microbiological parameters of untreated groundwater drawn from different types of wells, and the antimicrobial susceptibility profile of the bacteria isolated from this water. Wellwater samples were collected in two Brazilian cities. Although physical and chemical parameters of the water were suitable for drinking, Escherichia coli was detected in 33% of the samples. E. coli contaminated 65% of dug wells and 10.25% of drilled wells. Many bacteria isolated were resistant to multiple antibacterial agents, including β-lactams. Microbial contamination of this water was related to the well depth, and was more common in dug wells, making this water unfit for human consumption. Consumption of such contaminated and untreated water is a public health concern. Thus, individuals who regularly use such water must be alerted so they may either take preventive measures or connect to the water distribution system operated by local utilities.

Cytotoxicity, Genotoxicity, Antioxidant Potential and Chemical Composition of Leaves of Campomanesia pubescens (Mart. ex DC.) O.Berg

BACKGROUND Plants of the genus Campomanesia belong to the family Myrtaceae and are very abundant in Cerrado areas. Teas from leaves of species of this genus are used for intestinal infections, combating obesity, stomach pathology, fever and among others. OBJECTIVE The present study evaluated the chemical composition and antioxidant potential, cytotoxicity and genotoxicity of ethanolic extract from leaves of Campomanesia pubescens (Mart. ex DC.) O.Berg. METHOD For the evaluation of antioxidant activity, the free radical DPPH and for determination of phenolic compounds Folin-Ciocalteau reagent were used. Identification of the substances was performed by HPLC-DAD by comparison of the retention times with standards analyzed under the same conditions and by evaluation of molecular absorption spectra in the ultraviolet and visible region. The cytotoxicity, genotoxicity were evaluated using Allium cepa bioassay. RESULTS In the ethanolic extract 7-hydroxy-6-methyl-5-methoxyflavanone, 5,7-dihydroxy-6-methylflavanone, 5,7-dihydroxy-8-methylflavanone, 2´,4´-dihydroxy-6´-methoxychalcone, 5,7-dihydroxy-6,8- dimethylflavanone, 2´,4´-dihydroxy-5´-methyl-6´-methoxychalcone and 2´,4´-dihydroxy-3´,5´-dimethyl-6´- methoxychalcone were identified. The extract showed antioxidant activity and cytotoxic effects on cell division and increased chromosomal alterations in Allium cepa test. CONCLUSION These results showed antioxidant activity and suggest the cytotoxic and genotoxic effects in Allium cepa of ethanolic extract obtained from the leaves of Campomanesia pubescens.

Identification of microbiological contamination and mutagenic potential of surface waters of the municipality of Dourados, MS

The aim of the study was to evaluate the water quality of the Agua Boa stream and the lakes of the parks Arnulpho Fioravante and Antenor Martins (Dourados - MS), through physico-chemical parameters, microbiological analysis and mutagenicity test to monitor interference from Antropic activities and water depuration capacity. Initially the water samples were processed for coliform analysis using the Most Probable Number (MPN) technique. For the research of Salmonella spp., it was performed the pre-enrichment and enrichment of the water sample and streaking in Xylose Lysine Deoxicolate (XLD) agar. For the evaluation of the mutagenicity, smear peripheral blood of the specimens Astyanax lacustris were collected from the places studied. The Arnulpho Fioravante Park had higher mean values for Electrical Conductivity (353.5 uS cm -2 ), Total Dissolved Solids (179.3 mg L -1 ) and the highest values for total coliforms. Salmonella spp. it was identified in the water in both parks. The highest numbers of micronucleus were observed in Agua Boa stream. Considering microbiological contamination and higher micronucleus number, it’s relevant to highlight the effects of anthropic interferences in local biota. Therefore, it’s of great importance to intensify the supervision of the municipal legislation, in order to conserve and preserve these ecosystems

Naturalized Breeds in Brazil: Reports on the Origin and Genetic Diversity of the Pantaneiro Sheep

Brazil has several breeds of sheep, including animals that developed from breeds brought by settlers soon after their discovery. Over the years, these animals were under the process of natural selection, resulting in breeds that are considered naturalized. The Pantaneiro sheep shows rusticity and ability of adaptation to tropical climate regions and tolerance or resistance to disease and parasites. Molecular tools have marked the discovery of the origin and domestication processes of a wide variety of species, using both nuclear and mitochondrial molecular markers. These tools have aided in the understanding of evolutionary relationships, taxonomies, and demographics of various species and provided support to identify the most important areas for conservation programs, in addition to assisting in the analysis of genetic diversity in domestic, wildlife and endangered species. Researches using these tools show the importance of exploiting the potential of the genetic diversity found in locally adapted livestock. So far, a few studies were performed to observe that Pantaneiro sheep served as maternal basis for the origin of other breeds reared. Moreover, it is possible to suggest an European origin for the sheep populations studied; therefore, more studies using more markers are needed, so that it is possible to prove their origin.