Mutagenesis and Resistance Development of Bacteria Challenged by Silver Nanoparticles.

Because of their extremely broad spectrum and strong biocidal power, nanoparticles of metals, especially silver (AgNPs), have been widely applied as effective antimicrobial agents against bacteria, fungi, and so on. However, the mutagenic effects of AgNPs and resistance mechanisms of target cells remain controversial. In this study, we discover that AgNPs do not speed up resistance mutation generation by accelerating genome-wide mutation rate of the target bacterium Escherichia coli. AgNPs-treated bacteria also show decreased expression in quorum sensing (QS), one of the major mechanisms leading to population-level drug resistance in microbes. Nonetheless, these nanomaterials are not immune to resistance development by bacteria. Gene expression analysis, experimental evolution in response to sublethal or bactericidal AgNPs treatments, and gene editing reveal that bacteria acquire resistance mainly through two-component regulatory systems, especially those involved in metal detoxification, osmoregulation, and energy metabolism. Although these findings imply low mutagenic risks of nanomaterial-based antimicrobial agents, they also highlight the capacity for bacteria to evolve resistance.

Mutagenicity of a novel 2-phenylbenzotriazole (non-chlorinated 2-phenylbenzotriazole-9) in mice.

Dinitrophenylazo dyes can form 2-phenylbenzotriazoles (PBTAs) in the textile dyeing process upon the addition of chemical reducing agents. Some dinitrophenylazo dyes, as well as their respective reduced (non-chlorinated) and chlorinated PBTAs, are now found in rivers owing to wastewater from textile plants. This study aimed to investigate the genotoxicity of a new PBTA derived from C.I. Disperse Violet 93 azo dye, namely non-Cl PBTA-9. Primary DNA damage in the blood, liver, and colon cells, micronucleated cells in the bone marrow, and gene expression (NAT2, CYP1A1, TRP53, and CDKN1A) in liver cells were observed in mice, at acute oral exposure (gavage) doses of 5, 50, and 500 µg/kg body weight (b.w.). The non-chlorinated PBTA-9 caused DNA damage in the blood and liver (at 500 µg/kg b.w.) and in colon cells (at 5, 50, and 500 µg/kg), and increased the frequency of micronucleated cells in the bone marrow (at 5 and 50 µg/kg). No histological alterations or gene expression changes were observed. In conclusion, in vivo exposure to non-chlorinated PBTA-9 induced genetic damage in various rodent tissues, corroborating results previously obtained from the Ames test. Because this compound has been detected in rivers, exposure to humans and biota is a major concern.

No evidence for genotoxicity in mice due to exposure to intermediate-frequency magnetic fields used for wireless power-transfer systems.

Time varying magnetic fields (MFs) are used for the wireless power-transfer (WPT) technology. Especially, 85 kHz band MFs, which are included in the intermediate frequency (IF) band (300 Hz - 10 MHz), are commonly used WPT system for charging electric vehicles. Those applications of WPT technology have elicited public concern about health effects of IF-MF. However, existing data from health risk assessments are insufficient and additional data are needed. We assessed the genotoxic effects of IF-MF exposure on erythroid differentiation in mice. A high-intensity IF-MF mouse exposure system was constructed to induce an average whole-body electric field of 54.1 V/m. Blood samples were obtained from male mice before and after a 2-week IF-MF exposure (1 h/day, total: 10 h); X-irradiated mice were used as positive controls. We analyzed the blood samples with the micronucleus (MN) test and the Pig-a mutation assay. No significant differences were seen between IF-MF-exposed and sham-exposed mice in the frequencies of either MN or Pig-a mutations in mature erythrocytes and reticulocytes. IF-MF exposure did not induce genotoxicity in vivo under the study conditions (2.36× the basic restriction for occupational exposure, 22.9 V/m, in the International Commission on Non-Ionizing Radiation Protection (ICNIRP) guidelines). The absence of significant biological effects due to IF-MF exposure supports the practical application of this technology.

Bioflavonoids cause DNA double-strand breaks and chromosomal translocations through topoisomerase II-dependent and -independent mechanisms.

Bioflavonoids have a similar chemical structure to etoposide, the well-characterized topoisomerase II (Top2) poison, and evidence shows that they also induce DNA double-strand breaks (DSBs) and promote genome rearrangements. The purpose of this study was to determine the kinetics of bioflavonoid-induced DSB appearance and repair, and their dependence on Top2. Cells were exposed to bioflavonoids individually or in combination in the presence or absence of the Top2 catalytic inhibitor dexrazoxane. The kinetics of appearance and repair of γH2AX foci were measured. In addition, the frequency of resultant MLL-AF9 breakpoint cluster region translocations was determined. Bioflavonoids readily induced the appearance of γH2AX foci, but bioflavonoid combinations did not act additively or synergistically to promote DSBs. Myricetin-induced DSBs were mostly reduced by dexrazoxane, while genistein and quercetin-induced DSBs were only partially, but significantly, reduced. By contrast, luteolin and kaempferol-induced DSBs increased with dexrazoxane pre-treatment. Sensitivity to Top2 inhibition correlated with a significant reduction of bioflavonoid-induced MLL-AF9 translocations. These data demonstrate that myricetin, genistein, and quercetin act most similar to etoposide although with varying Top2-dependence. By contrast, luteolin and kaempferol have distinct kinetics that are mostly Top2-independent. These findings have implications for understanding the mechanisms of bioflavonoid activity and the potential of individual bioflavonoids to promote chromosomal translocations. Further, they provide direct evidence that specific Top2 inhibitors or targeted drugs could be developed that possess less leukemic potential or suppress chromosomal translocations associated with therapy-related and infant leukemias.

The mutagenesis moonshot: The propitious beginnings of the environmental mutagenesis and genomics society.

A mutagenesis moonshot addressing the influence of the environment on our genetic wellbeing was launched just 2 months before astronauts landed on the moon. Its impetus included the discovery that X-rays (Muller HJ. [1927]: Science 64:84-87) and chemicals (Auerbach and Robson. [1946]: Nature 157:302) were germ-cell mutagens, the introduction of a growing number of untested chemicals into the environment after World War II, and an increasing awareness of the role of environmental pollution on human health. Due to mounting concern from influential scientists that germ-cell mutagens might be ubiquitous in the environment, Alexander Hollaender and colleagues founded in 1969 the Environmental Mutagen Society (EMS), now the Environmental Mutagenesis and Genomics Society (EMGS); Frits Sobels founded the European EMS in 1970. As Fred de Serres noted, such societies were necessary because protecting populations from environmental mutagens could not be addressed by existing scientific societies, and new multidisciplinary alliances were required to spearhead this movement. The nascent EMS gathered policy makers and scientists from government, industry, and academia who became advocates for laws requiring genetic toxicity testing of pesticides and drugs and helped implement those laws. They created an electronic database of the mutagenesis literature; established a peer-reviewed journal; promoted basic and applied research in DNA repair and mutagenesis; and established training programs that expanded the science worldwide. Despite these successes, one objective remains unfulfilled: identification of human germ-cell mutagens. After 50 years, the voyage continues, and a vibrant EMGS is needed to bring the mission to its intended target of protecting populations from genetic hazards. Environ. Mol. Mutagen. 61:8-24, 2020. © 2019 Wiley Periodicals, Inc.

Twenty Years of Reflections in Mutation Research.

Reflections is a component of Mutation Research Reviews devoted to historical and philosophical themes pertaining to the subject of mutation. Reflections was initiated in 1999 and has included a broad array of topics centered on mutation research, but overlapping other scientific fields and touching upon history, sociology, politics, philosophy and ethics. This commentary offers an editor's reflections on the 44 papers in the Reflections series, including the people who contributed to the series and the topics that they discussed.

Mutagenic effects of spent potliner and derivatives on Allium cepa L. and Lactuca sativa L.: A molecular approach.

Spent potliner (SPL) is a solid residue generated by the aluminum industry. Its composition is variable and complex, containing fluoride and cyanide salts as well as aluminum, which contributes to its toxicity. SPL is sometimes released directly into the soil, where it is prone to leaching and has the potential to cause alterations and damage to DNA. Considering that polymorphism analysis of simple sequence repeat (SSR) and inter-simple sequence repeat (ISSR) DNA markers is an interesting tool to determine the mutagenicity of an environmental pollutant, the present study adopted this approach to verify the mutagenic potential of SPL and its main toxic components (aluminum, fluoride, and cyanide) on root tip cells of Lactuca sativa and Allium cepa. Alterations in ISSR and SSR regions were identified by DNA fingerprinting (gain and loss of bands and changes in band intensity). The estimated dissimilarities indicated differences between treatments and the negative control. Furthermore, the relationship between the amplification profile of the markers and alterations in cell mitosis was discussed.

The Glyphosate-Based Herbicide Roundup Does not Elevate Genome-Wide Mutagenesis of Escherichia coli.

Mutations induced by pollutants may promote pathogen evolution, for example by accelerating mutations conferring antibiotic resistance. Generally, evaluating the genome-wide mutagenic effects of long-term sublethal pollutant exposure at single-nucleotide resolution is extremely difficult. To overcome this technical barrier, we use the mutation accumulation/whole-genome sequencing (MA/WGS) method as a mutagenicity test, to quantitatively evaluate genome-wide mutagenesis of Escherichia coli after long-term exposure to a wide gradient of the glyphosate-based herbicide (GBH) Roundup Concentrate Plus. The genome-wide mutation rate decreases as GBH concentration increases, suggesting that even long-term GBH exposure does not compromise the genome stability of bacteria.

The influence of heavy metals on toxicogenetic damage in a Brazilian tropical river.

Anthropogenic activities in tropical rivers favor the eutrophication process, which causes increased concentration of heavy metals. The presence and bioaccumulation of metals are directly related to the presence of genotoxic damage in aquatic organisms. Thus, we evaluated the presence of heavy metals (Fe, Zn, Cr, Cu and Al) and performed toxicogenetic tests in surface (S) and bottom (B) of water samples of the Poti river (Piaui/Brazil). Cytotoxicity and genotoxicity tests were performed in Allium cepa, and micronucleus (MN) and comet assay were performed in Oreochromis niloticus. The chemical analysis showed concentrations above the limit for Cu, Cr, Fe and Al according to Brazilian laws, characterizing anthropogenic disturbance in this aquatic environment. Toxicogenetic analysis presented significant cytotoxic, mutagenic and genotoxic effects in different exposure times and water layers (S and B), especially alterations in mitotic spindle defects, MN formations, nuclear bud and DNA strand breaks. Correlations between Fe and cytotoxicity, and Al and mutagenicity were statistically significant and point out to the participation of heavy metals in genotoxic damage. Therefore, Poti river water samples presented toxicogenetic effects on all bioindicators analyzed, which are most likely related to heavy metals pollution.

Cytotoxicity and genotoxicity of Guaribas river water (Piauí, Brazil), influenced by anthropogenic action.

In general, tropical rivers have a great impact on human activities. Bioaccumulation of toxins is a worldwide problem nowadays and has been, historically, overlooked by the supervisory authorities. This study evaluated cytogenotoxic effects of Guaribas river (a Brazilian river) water during dry and rainy seasons of 2014 by using the Allium cepa test system. The toxicogenetic variables, including root growth, mitotic index, and chromosomal aberrations, were analyzed in meristematic cells of A. cepa exposed to water samples taken from the up-, within, and downstream of the city Picos (state: Piauí). The physical-chemical parameters were also analyzed to explain water quality and possible anthropogenic action. Additionally, the presence of heavy metals was also analyzed to explain water quality and possible damaging effects on eukaryotic cells. The results suggest that the river water exerted cytotoxic, mutagenic, and genotoxic effects, regardless of the seasons. In addition, Guaribas river presented physico-chemical values outside the Brazilian laws, which can be a characteristic of human pollution (domestic sewage, industrial, and local agriculture). The genetic damage was positively correlated with higher levels of heavy metals. The pollution of the Guaribas river water may link to the chemical contamination, including the action of heavy metals and their impacts on genetic instability in the aquatic ecosystem. In conclusion, necessary steps should be taken into account for further toxicogenetic studies of the Guaribas river water, as it has an influence in human health of the same region of Brazil.

Citotoxicidade das águas do Rio do Peixe (São Paulo-Brasil), em células meristemáticas de raiz de Allium cepa L / Cytotoxicity of waters of the River Peixe (São Paulo-Brazil), in meristematic root cells of Allium cepa L

Nas últimas décadas, a qualidade da água do Rio do Peixe, localizado no interior do Estado de São Paulo, está diminuindo, principalmente porque o rio vem sofrendo com o desmatamento feito às suas margens, o que provocou o assoreamento e permitiu o despejo de todos os tipos de resíduos, em especial das indústrias que ficam ao seu redor. Assim, o objetivo deste trabalho foi avaliar o potencial citotóxico das águas do Rio do Peixe, coletadas em quatro propriedades rurais localizadas em Garça­SP (nascente), Tupã-SP, Flórida Paulista-SP e Ouro Verde-SP (jusante), usando como sistema teste vegetal as células meristemáticas de raiz de Allium cepa L. Os resultados obtidos mostraram que todas as amostras das águas coletadas não apresentaram efeito citotóxico após 24 horas de tratamento. Entretanto, as amostras obtidas em Tupã, Flórida Paulista e Ouro Verde (jusante) foram mutagênicas, pois causaram o aparecimento de alterações nas células, principalmente, dos tipos metáfases-colchicínicas, anáfases desorganizadas, multipolares e com ponte cromatídica, além de células micronucleadas. Portanto, os resultados obtidos alertam para o perigo eminente que os efluentes lançados indiscriminadamente no ambiente, de rios e lagos, podem representar não só aos organismos que ali habitam, mas a todos os outros que dele dependem, seja para lazer, limpeza, higiene ou alimentação.

(Cytotoxicity of waters of the River Peixe (São Paulo-Brazil), in meristematic root cells of Allium cepa L.) In recent decades, the water quality of the River Peixe, located within the State of São Paulo, has been decreasing, mainly because the river has suffered from deforestation made to its banks, causing silting and allowed the dumping of all types of waste, especially in industries that are around you. The objective of this study was to evaluate the cytotoxic potential of the waters of the River Peixe, collected at four farms located in Garça-SP (source), Tupã-SP, Flórida Paulista-SP and Ouro Verde-SP (downstream), using as a system test plant root meristematic cells of Allium cepa L. The results showed that all water samples collected was no cytotoxic effect after 24 hours of treatment. However, samples taken in Tupã, Flórida Paulista and Ouro Verde (downstream) were mutagenic, because it caused the appearance of changes in cells, mainly types of colchicine-metaphase, disorganized anaphase, multipolar and with chromatid bridge, addition of micronucleated cells. Therefore, the results warn of imminent danger that the effluents discharged indiscriminately into the environment may represent not only the organisms that live there, but for all others who depend on it, whether for leisure, cleaning, hygiene or food.

Greetings from The International Association of Environmental Mutagenesis and Genomics Societies.

The International Association of Environmental Mutagenesis and Genomics Societies (IAEMGS) is an organization that promotes basic and applied research on environmental mutagenesis and genomics. In this article, as President of IAEMGS, I stress the important role of Genes and Environment to spread the voice of Asia to the international scientific community. Open access will support the journal in achieving this mission.

Vegetal test-system investigation on cytotoxicity of water from urban streams located in the northeastern region of Maringá, Paraná State, Brazil / Investigação da citotoxicidade das águas de córregos urbanos localizados na região nordeste de Maringá, Estado do Paraná, Brasil, em sistema-teste vegetal

The increase in consumption of water, the destruction of riparian forests and the pollution caused by humans, have severely degraded several water resources. Numerous stream crisscross the city of Maringá, Paraná state, Brazil and most of their sources are scattered through the urban region. Current analysis assesses the cytotoxic potential of water from the Corregozinho, Isalto, Morangueira and Ozório streams, located within the high-populated northeastern region of Maringá, inside the urban perimeter. Root meristematic cells of Allium cepa were used as test-system. The roots of onion were prepared by Feulgen's reaction and stained with Schiff's reagent. Results showed that there were no statistically significant changes evaluated by the chi-square test on the rates of cell division in cells of Allium cepa roots treated with the water of the streams when compared to data from controls. However, further analyses should be undertaken at different times for a periodic assessment of conditions in the streams, coupled with an awareness of the population on the environment.

O aumento do consumo da água, a devastação das matas ciliares e a poluição causada pela espécie humana, fizeram com que muitos recursos hídricos fossem drasticamente degradados. A cidade de Maringá, Paraná, Brasil possui inúmeros córregos que a cortam, sendo que as nascentes da maioria deles estão espalhadas pela região urbana. Assim, este trabalho objetivou avaliar o potencial citotóxico das águas dos córregos, Corregozinho, Isalto, Morangueira e Ozório, localizados na região nordeste de Maringá, no perímetro urbano, local de grande concentração da população urbana. Foram utilizadas como sistema-teste as células meristemáticas de raiz de Allium cepa. As raízes de cebola foram preparadas pela reação de Feulgen e coradas com o reativo de Schiff. Os resultados obtidos indicaram que não houve alterações, estatisticamente significativas, pelo teste do qui-quadrado, nos índices de divisão celular nas células das raízes de Allium cepa tratadas com as águas dos córregos, em relação aos dados obtidos para os controles. Entretanto, convém que outras análises, em diferentes épocas, sejam realizadas, para uma avaliação periódica das condições apresentadas por estes córregos, e aliado a estes resultados, conscientizar a população em relação aos danos provocados ao meio ambiente e a necessidade de conservação do mesmo.

Mutagénesis ambiental y el uso de biomarcadores para predecir el riesgo de cáncer: [revisión] / Environmental mutagenesis and use of biomarkers in cancer risk prediction: [review]

The field of environmental mutagenesis or toxicology genetics aims to study the genetic damage that leads to mutations produced by physical, chemical and biological agents, to identify these agents and analyze their interactions and ways of action. There are enough experimental and epidemiological evidences implicating mutations in oncogenes, tumor suppressor genes and DNA repair genes as determinants in the onset and progression of the neoplastic process. A valuable tool in public and occupational health is the monitoring of populations exposed to potentially hazardous agents. The objective is to protect the health and quality of life of high risk groups on account of the nature of the agents of exposure. Monitoring of genotoxic effects in exposed populations as well as the analysis of susceptibility polymorphism are visualized as key tools in the realm of future public and occupational health in order to prevent the occurrence of environmental and specially occupational origin of tumors. This paper reviews the main concepts concerning this issue and refers to studies on the subject in Costa Rica.