Genotoxic and mutagenic effects of polluted surface water in the midwestern region of Brazil using animal and plant bioassays

Abstract This study aimed to evaluate DNA damage in animal and plant cells exposed to water from the Água Boa stream (Dourados, Mato Grosso do Sul, Brazil) by using bioassays, and to identify the chemical compounds in the water to determine the water quality in the area. Through the cytotoxicity bioassay with Allium cepa, using micronucleus test, and comet assay, using Astyanax altiparanae fish, the results indicated that biological samples were genetically altered. Micronuclei were observed in erythrocytes of A. altiparanae after exposure to water from locations close to industrial waste discharge. The highest DNA damage observed with the comet assay in fish occurred with the exposure to water from locations where the presence of metals (Cu, Pb, Cd, Ni) was high, indicating the possibility of genotoxic effects of these compounds. Thus, these results reinforce the importance of conducting genotoxicity tests for developing management plans to improve water quality, and indicate the need for waste management before domestic and industrial effluents are released into the rivers and streams.

Efficiency analysis of the australian wastewater treatment system in a pig slaughterhouse / Análise da eficiência do sistema australiano de tratamento de águas residuais em um abatedouro de suínos

Pig farming is a significant economic activity in the food industry and the meat trade. However, pig slaughterhouses release their waste through effluents, which may spread pathogenic microorganisms and degrade the environment, particularly in bodies of water. The aim of the present study was to assess the efficiency of the Australian wastewater treatment system and the quality of the Effluent Treatment Station (ETS) of a pig slaughterhouse, which flows into the Laranja Azeda stream in the city of Dourados (State of Mato Grosso do Sul, Brazil). Effluentsampling was carried out in the ETS using treatment ponds (Australian system), from the input to the output, over a period of one year. Physicochemical and biological parameters were assessed to measure the effluentquality. The results showed a reduction in organic matter and microorganisms. The concentration of dissolved oxygen, pH, effluent temperature, redox potential, as well as the amount of coliforms, exhibited a significant reduction (p<0.005). The waste management brought by the ETS promoted the removal of pollutants from the effluent. However, these procedures were not enough to satisfy Brazilian and international parameters proposed by the WHO, which regulate the dumping of waste into water bodies.

A suinocultura é uma atividade econômica que ocupa posição de destaque no cenário mundial alimentício vinculado ao comércio da carne. No entanto, abatedouros de suínos geram resíduos que são liberados pelos efluentes e podem disseminar microrganismos patogênicos e degradar o meio ambiente, especialmente em corpos hídricos. O objetivo do estudo foi avaliar a eficiência do sistema australiano e qualidade do efluente proveniente da Estação de Tratamento de Efluente (ETE) de um abatedouro de suínos liberada no Córrego Laranja Azeda localizado na cidade de Dourados (Estado do Mato Grosso do Sul, Brazil). As coletas das amostras de efluentes na ETE foram realizadas nas lagoas de tratamento (sistema australiano) desde a sua entrada até a saída durante o período de um ano. Para mensuração da qualidade de água foram avaliados parâmetros físico-químicos e biológicos. Parâmetros referentes à Demanda Bioquímica de Oxigênio e a Demanda Química de Oxigênio foram aferidos de acordo com a American Public Health Association e, além destes foram mensurados quantidade de oxigênio dissolvido, pH, temperatura, condutividade elétrica, sólidos totais dissolvidos e potencial de oxi-redução do efluente. A técnica dos tubos múltiplos foi utilizada para quantificação de coliformes totais (CT) e coliformes termotolerantes (CTT). Os resultados apontaram a redução de matéria orgânica e microrganismos. As médias dos parâmetros avaliados apresentaram redução significativa (p<0,005) para a concentração de oxigênio dissolvido, pH, temperatura da água, potencial de oxi-redução, quantidade de coliformes presentes na água. O manejo dos resíduos gerados pela ETE acarretou na remoção de poluentes do efluente, mas não foi eficiente em atender os parâmetros legais brasileiros e internacionais que normatizam o seu despejo em corpos de água. Considera-se necessária maior fiscalização das condições referentes a qualidade de água do efluente lançado neste córrego, pois o resultado indica risco de saúde para a comunidade ribeirinha que faz uso dessa água para banhos, dessedentação de animais, irrigação e outras atividades domésticas.

Novel point mutations in the ERG11 gene in clinical isolates of azole resistant Candida species

The azoles are the class of medications most commonly used to fight infections caused by Candida sp. Typically, resistance can be attributed to mutations in ERG11 gene (CYP51) which encodes the cytochrome P450 14α-demethylase, the primary target for the activity of azoles. The objective of this study was to identify mutations in the coding region of theERG11 gene in clinical isolates of Candidaspecies known to be resistant to azoles. We identified three new synonymous mutations in the ERG11 gene in the isolates of Candida glabrata (C108G, C423T and A1581G) and two new nonsynonymous mutations in the isolates of Candida krusei - A497C (Y166S) and G1570A (G524R). The functional consequence of these nonsynonymous mutations was predicted using evolutionary conservation scores. The G524R mutation did not have effect on 14α-demethylase functionality, while the Y166S mutation was found to affect the enzyme. This observation suggests a possible link between the mutation and dose-dependent sensitivity to voriconazole in the clinical isolate of C. krusei. Although the presence of the Y166S in phenotype of reduced azole sensitivity observed in isolate C. kruseidemands investigation, it might contribute to the search of new therapeutic agents against resistant Candida isolates.

Genotipo, sensibilidad a los antifúngicos y formación de biofilm de aislados de Trichosporon asahii recuperados de la orina de pacientes hospitalizados / Genotype, antifungal susceptibility, and biofilm formation of Trichosporon asahii isolated from the urine of hospitalized patients

En este estudio fueron analizadas mediante el cultivo muestras de orina de pacientes hospitalizados en la región centro-oeste de Brasil; los microorganismos aislados fueron identificados filogenéticamente como Trichosporon asahii. A través del análisis de máxima parsimonia de las secuencias de IGS1, fueron encontrados 3 genotipos que no habían sido descritos anteriormente. Las concentraciones inhibitorias mínimas frente a los 9 aislados identificados presentaron un rango de 0,06-1µg/ml en el caso de la anfotericina B, de 0,25-4µg/ml en el del fluconazol, y de 0,03-0,06µg/ml en el del itraconazol. Aproximadamente 6/9 de los aislados de T. asahii formaron biopelículas en la superficie de microplacas de poliestireno. Este trabajo documenta el aislamiento de T. asahii como agente causal de infeciones urinarias nosocomiales. Además, demuestra que la región IGS1 puede ser considerada una nueva herramienta epidemiológica para la genotipificación de los aislados de T. asahii. Los genotipos menos comunes encontrados en este estudio pueden estar relacionados con las características epidemiológicas locales

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 0.06­1µg/ml for amphotericin B, 0.25­4µg/ml for fluconazole, and 0.03­0.06µg/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

Application of microsatellite markers for breeding and genetic conservation of herds of Pantaneiro sheep

Background The aim of the present study was to assess the genetic diversity of Pantaneiro sheep, using microsatellite markers, in order to assist maintenance and management plans, enhance mating systems and reduce the inbreeding rate. A total of 127 animals were genotyped at eight microsatellite loci. They belonged to populations from the Experimental Farm of the Universidade Federal da Grande Dourados (UFGD) (Dourados/MS/Brazil) and Embrapa Pantanal (Corumbá/MS/Brazil). Results The population of Pantaneiro sheep from the UFGD exhibited a high mean number of alleles (11.13) and allelic richness (10.66). The polymorphic information content was highly informative in the locus studied, resulting in a mean value of 0.71. Observed heterozygosity was lower than expected for all molecular markers assessed. The analysis of molecular variance showed a differentiation rate of 5.2% between populations. Conclusions The results of the statistical parameters indicated that populations of Pantaneiro sheep require special attention on herd management, and it's further necessary to implement breeder exchange programs in order to preserve the genetic variability of these populations. Furthermore, the maintenance of those populations in their typical habitats is rather required to allow different responses from the herds to the interactions between genotype and environment.

Genetic identification of interspecific hybrid of Neotropical catfish species (Pseudoplatystoma corruscans vs. Pseudoplatystoma reticulatum) in rivers of Mato Grosso do Sul State, Brazil

Hybrids of the species Pseudoplatystoma corruscans vs. Pseudoplatystoma reticulatum are extensively traded because they are more docile and precocious compared to their parents. This study aimed to identify the occurrence of hybrid 'Surubim' in rivers of the State of Mato Grosso do Sul, applying molecular techniques. Pseudoplatystoma corruscans, Pseudoplatystoma reticulatum and hybrid 'Surubim' were detected using multiplex PCR and PCR-RFLP in RAG2, GLOBIN, EF1α, 18S rRNA nuclear genes, and mitochondrial 16S rRNA gene. The results revealed the presence of F1 hybrids in rivers and the occurrence of genetic introgression, evidenced by the Post-F1 hybrids. Furthermore, discrepancies between the morphological and genetic identification were observed, demonstrating the effectiveness of using different molecular markers for the correct identification of these hybrids. The necessity of implementation of management and conservation projects to maintain the genetic integrity of native species populations threatened by hybridization in the rivers of Mato Grosso do Sul State is evident....

Híbridos envolvendo as espécies Pseudoplatystoma corruscans vs. Pseudoplatystoma reticulatum são amplamente comercializados por apresentarem maior docilidade e precocidade quando comparados aos parentais. Neste trabalho objetivou-se identificar, por meio de técnicas moleculares, a existência de surubins híbridos em rios do estado do Mato Grosso do Sul. Pseudoplatystoma corruscans, Pseudoplatystoma reticulatum e 'Surubim' híbrido foram identificados utilizando as técnicas de PCR multiplex e PCR-RFLP dos genes nucleares RAG2, GLOBINA, EF1α, 18S rRNA e mitocondrial 16S rRNA. Os resultados demonstraram a presença de híbridos F1 em rios e a ocorrência da introgressão genética, evidenciada pelos híbridos Pós-F1. Além disso, foram observadas discrepâncias entre a identificação morfológica e genética, demonstrando a eficácia da utilização de diversos marcadores moleculares para identificação correta destes híbridos. Torna-se evidente a necessidade da implantação de projetos de manejo e conservação, a fim de manter a integridade genética de populações de espécies nativas em rios do estado do Mato Grosso do Sul, que encontram-se ameaçadas pela hibridação...

Genetic variability of Herpailurus yagouaroundi, Puma concolor and Panthera onca (Mammalia, Felidae) studied using Felis catus microsatellites

We used four microsatellite loci (Fca08, Fca45, Fca77 and Fca96) from the domestic cat, Felis catus, to investigate genetic variability in specimens of Herpailurus yagouaroundi (jaguarundi, otter cat, eyra), Puma concolor (cougar, mountain lion, puma) and Panthera onca (jaguar) held in various Brazilian zoos. Samples of DNA from the cats were PCR amplified and then sequenced before being analyzed using the CERVUS program. Our results show a mean polymorphic information content (PIC) of 0.83 for H. yagouaroundi, 0.66 for P. concolor and 0.69 for P. onca and a mean of 10.3 alleles for the Fca08 locus, 5.3 for Fca 45, 9 for Fca 77 and 14 for Fca 96. These results indicate a relatively high level of genetic diversity for the specimens studied.