Acute and subchronic effects of petroleum on the freshwater fish Hoplias aff. malabaricus

Petroleum water soluble fraction (WSF) impairs organisms, but damages may vary among cell and tissue levels. The aim of the present study was to evaluate the acute (24 h, 48 h, 72 h) and subchronic effects (36 days) of WSF (0%, 25% and 100%) in juveniles of the Neotropical top predator fish Hoplias aff. malabaricus. The effects of WSF were evaluated at a molecular level using the comet assay and micronucleus test for genome damage; and at a morphological level through histological identification of liver pathologic lesions. In both acute and subchronic exposure we found low levels of DNA damage (< 10% of comet tail) and non-significant frequency of micronucleus in WSF exposed fish. The most significant liver lesions in WSF exposed fish were fatty vacuolization, hypertrophy and focal necrosis. Since these tissue injuries were progressive and persistent, their irreversibility may negatively affect fish recruitment, even in a such resistant top predator.

Biomonitoring of air quality in the Bodoquena microregion, Mato Grosso Do Sul: mutagenic and morphoanatomical alterations in Tradescantia pallida (rose) D.R. Hunt var. purpurea

The objective of this study was to assess air quality in relation to vehicular traffic flow in cities located at different elevations in the Bodoquena microregion, state of Mato Grosso do Sul, Brazil. To do so, a micronucleus test was carried out using the TRAD-MCN bioassay on young Tradescantia buds collected from February to November 2018 in seven cities of the microregion with different traffic flow intensities. Meteorological parameters were evaluated, and vehicular traffic was counted to determine traffic flow in each city. With data from the Shuttle Radar Topography Mission (SRTM) and processing in Esri ArcGIS® software version 10.5.1, the regions was mapped based on an Elevation Model. Morphoanatomical analyses were performed according to standard methodology. Measurements were taken of thickness, length and width of tissues and structures, including the upper and lower cuticle, upper and lower epidermis, hypodermis and mesophyll. The greatest traffic flow was found in the cities of Bodoquena, Guia Lopes da Laguna, Jardim, and Porto Murtinho, with the period from 5:00 to 6:00 p.m. showing the highest traffic flow. The greatest frequency of mutagenic alterations was found in the city of Guia Lopes da Laguna, although the results did not differ significantly from Bonito, Caracol, and Jardim. Throughout the biomonitoring, the summer and autumn seasons showed the greatest micronuclei frequencies in all evaluated cities. Variations in the tissue/structure thickness was observed across cities and seasons, but with a decrease in thickness during autumn. In general, the tissues/structures were smaller for the cities of Nioaque and Porto Murtinho, while the anatomical and morphological characteristics of leaf length and thickness showed no differences among cities. We found limited correlation between micronuclei frequency and traffic flow, supporting the hypothesis that although mutagenic alterations are observed in T. pallida, in this microregion the changes are numerically lower when compared to other regions of the state. In light of the genotoxic and morphoanatomical factors assessed herein, the Bodoquena microregion appears to be well preserved in terms of air quality, presenting low micronuclei frequency and a limited reduction in tissues and leaf structures, regardless of the season.

Evaluation of Subchronic Toxicity and Genotoxicity of Ethanolic Extract of Aster glehni Leaves and Stems

Aster glehni, a traditional plant on Ulleung Island in the Republic of Korea, has been recognized for its multiple medicinal properties. However, potential toxicity and safety analyses of A. glehni have not been previously investigated. Therefore, this study aimed to evaluate the safety profile of ethanolic extract of A. glehni leaves and stems (EAG) in terms of genotoxicity and subchronic oral animal toxicity under OECD guidelines and GLP conditions. Toxicological assessments were performed at doses of 1,250, 2,500, and 5,000 mg/kg/day in a 13-week oral repeated-dose toxicity study of EAG in male and female SD rats. In addition, an Ames test, an in vitro mammalian chromosomal aberration test, and a micronucleus test were performed. No toxicological changes in clinical signs, body weights, water and food consumption, urinalysis, hematology, clinical biochemistry, gross findings, and histopathological examinations were observed in subchronic oral animal toxicity. In addition, EAG gave negative results when evaluated using in vitro and in vivo genotoxicity tests. In conclusion, the no-observed-adverse-effect level (NOAEL) of EAG was considered to be 5,000 mg/kg/day, and no target organs were identified in both sexes of rats. EAG was also classified as nonmutagenic and nonclastogenic in genotoxicity testing. Collectively, these results show a lack of general toxicity and genotoxicity for EAG that supports clinical work for development as a herbal medicine.

ZINC CONTAMINATION IS AN UNDERESTIMATED RISK TO AMPHIBIANS: TOXICITY EVALUATION IN TADPOLES OF FEJERVARYA LIMNOCHARIS

Aquatic environments are often contaminated with zinc. Amphibian tadpoles are likely to be exposed to high concentrations of zinc present in these environments. We determined the acute and sub-chronic toxicity of ZnCl2 on Fejervarya limnocharis tadpoles under laboratory conditions. The LC50 values of ZnCl2 were found to be 5.81, 4.32, 3.79 and 3.61 mg/L at 24, 48, 72 and 96 h of exposure respectively. Long-term exposure to sub-lethal concentrations of ZnCl2 induced significant mortality in concentration and time dependent manner. Sub-lethal ZnCl2 exposure significantly altered survival, body length and body weight at metamorphosis. Micronucleus test and comet assay indicated the genotoxic potential of ZnCl2. Significant increase in DNA strand break was observed following ZnCl2 exposure equivalent to 1% of the of 24 h LC50 value. The findings indicate possible adverse to tadpoles inhabiting aquatic environments contaminated with zinc. In addition, the findings may be extrapolated to aquatic organisms of similar torphic status.

Cytogenetic effect of caffeine in the micronucleus test

Relevance. The consumption of caffeine-containing food in the modern world must necessarily be safe for humans, including should not affect the hereditary material of the body. Objective: to determine the possible effect of caffeine at the cytogenetic level by the micronucleus method on erythrocytes. Materials and Methods. The objects for the study were non-linear mice, which were divided into 6 groups - one control group and 5 experimental groups. The first experimental group and the second in the experiment received caffeine in doses of 40 mg/kg and 100 mg/kg.The control group received saline. Caffeine was administered orally. The mutagen (dioxidine) was injected intraperitoneally. On the 5th day of the experimental study, we performed blood sampling for cytogenetic analysis. Results and Discussion. Our study of the caffeine preparation made it possible to determine the following patterns. Firstly, when administered within 5 days, caffeine at a dose of 40 and 100 mg/kg did not cause an increase in the number of micronuclei in erythrocytes in mice. Secondly, the combined use of caffeine (both at a dose of 40mg/kgand at a dose of 100 mg / kg) and dioxidine significantly increased the level of micronuclei in comparison with the control group. Thirdly, caffeine at a dose of 40mg/kgdid not increase the mutagenic activity of dioxidine, but a dose of caffeine of 100mg/kgwhen combined with a mutagen led to a significant increase in the level of cytogenetic damage. Conclusion. According to our data, caffeine in the experimental study was not a mutagen, but at a dose of 100 mg/kg it represented a comutagenic effect.

Morphofunctional features of proliferating cells exposed to PSMA peptide

The effect of the synthetic PSMA peptide on dividing cells of laboratory animals was studied. The experiment was carried out on male white laboratory mice of the BALB/c-line. The toxic effect of PSMA peptidi was evaluated at therapeutic (1.4 μg / kg of animal weight or 0.04 μg / animal) and subtoxic (140 μg / kg of animal weight or 4.0 μg / animal) doses. The cytotoxic effect of PSMA peptide on red bone marrow cells and cambial intestinal cells of the of laboratory mice was determined. A decrease in the proliferative activity of the colon crypt cells was revealed upon administration of a subtoxic dose of the PSMA peptide and there were no signs of toxic damage to the red bone marrow cells of animals. Key words: toxicity, proliferation, synthetic peptides, mitotic index, micronucleus test.

PIG-A gene mutation as a genotoxicity biomaker in polycyclic aromatic hydrocarbon-exposed barbecue workers

Background The PIG-A gene mutation assay is a valuable tool for measuring in vivo gene mutations in blood cells. The human PIG-A assay, used as a potential genotoxicity biomarker, is minimally invasive, sensitive, and cost-efficient; however, the relationship between carcinogen exposure and PIG-A mutations is not well understood. Methods We investigated the genotoxic effect of red blood cells using PIG-A assay and lymphocyte cytokinesis-block micronucleus test in barbecue restaurant workers (N = 70) exposed to polycyclic aromatic hydrocarbons (PAHs) and self-identified healthy control subjects (N = 56). Urinary PAH metabolites were measured to evaluate internal exposure levels. Results Multivariate Poisson regression showed that the PAH-exposed workers exhibited significantly higher PIG-A mutant frequency (MF) (8.04 ± 6.81 × 10− 6) than did the controls (5.56 ± 5.26 × 10− 6) (RR = 0.707, 95% CI: 0.615–0.812, P < 0.001). These results indicate that PAH exposure is a risk factor for elevated PIG-A MF. The frequencies of micronuclei (MN) and nuclear buds (NBUD) in the PAH-exposed workers (MN: 3.06 ± 2.07 ‰, NBUD: 1.38 ± 1.02 ‰) were also significantly higher than in the controls (MN: 1.46 ± 0.64 ‰, P < 0.001; NBUD: 0.70 ± 0.60 ‰, P < 0.001). Additionally, PIG-A MFs showed better associations with several urinary hydroxylated PAH metabolites (P2-OH-Flu = 0.032, r2-OH-Flu = 0. 268; P2-OH-Phe = 0.022, r2-OH-Phe = 0.286; P3-OH-Phe = 0.0312, r3-OH-Phe = 0.270; P4-OH-Phe = 0.018, r4-OH-Phe = 0.296), while the increase in MN, NPB, and NBUD frequencies was not associated with any OH-PAH metabolites; and high-PAH-exposed workers showed the highest PIG-A MFs. Furthermore, there was a significant association between PIG-A MF and PAH exposure levels (Chi-square test for trend, P = 0.006). Conclusions Our results indicate that an increase in PIG-A MF in barbecue workers could reflect the response to PAH exposure, providing evidence of its potential as a genotoxicity biomarker in human risk assessment.

Assessment of genotoxicity of Ssanghwa-tang, an herbal formula, by using bacterial reverse mutation, chromosome aberration, and in vivo micronucleus tests

Objectives: Ssanghwa-tang (SHT) is a traditional herbal formula comprising nine medicinal herbs, and it is used for reducing fatigue in Korea. SHT exerts various effects such as anti-inflammatory, antioxidant, and anti-aging activities, and protection against acute hepatotoxicity. However, the genotoxicity of SHT has not yet been established.Methods: Ten components were identified in SHT water extract by using high-performance liquid chromatography analysis. We assessed the genotoxicity of SHT by using bacterial reverse mutation (Ames test), chromosome aberration, and in vivo micronucleus tests.Results: The contents of paeoniflorin, glycyrrhizin, and liquiritin apioside in SHT were 15.57, 6.94, and 3.48 mg/g extract, respectively. SHT did not increase the revertant colonies of Salmonella typhimurium and Escherichia coli strains in the presence or absence of metabolic activity. Although SHT did not induce structurally abnormal chromosomes in Chinese hamster lung (CHL) cells in the presence of metabolic activity, the number of structurally aberrated chromosomes increased dose-dependently in the absence of metabolic activity. In the in vivo micronucleus test, SHT did not affect the formation of micronuclei compared with the vehicle control.Conclusions: Genotoxicity of SHT was not observed in the Ames test and in vivo micronucleus test. However, based on the results of chromosome aberration test, it can be presumed that SHT has the potential to induce genotoxicity because it induced structurally abnormal chromosomes in the absence of metabolic activity.

Genotoxicity Assessment and Protective Effect of Anogeissus leiocarpus Roots against Cyclophosphamide-Induced DNA Damage In Vivo

Background. Belonging to the family of Combretaceae, the roots of Anogeissus leiocarpus are traditionally used to treat diabetes, wounds, infections, pain, and gastrointestinal diseases. To our knowledge, no genotoxicity assessment of the plant was reported. Hence, this study was designed to evaluate the potential genotoxic and protective effects of extract of Anogeissus leiocarpus roots using the micronucleus test on mice bone marrow cells in vivo. Methods. Three different concentrations (250, 500, and 1000 mg·kg−1) of hydroalcoholic extract of roots of A. leiocarpus were administered daily for 7 days per os to mice, and the genotoxicity was induced by the administration ip of cyclophosphamide. Genotoxicity and cytotoxicity were evaluated by counting, respectively, the number of micronucleated polychromatic erythrocytes and polychromatic erythrocytes to total erythrocytes in the bone marrow of mice. Results. The administration of A. leiocarpus did neither increase the ratio of the polychromatic erythrocyte (PCE) nor the frequency of micronucleated PCE (MNPCE) significantly in the bone marrow cells of the mice, compared to the vehicle control animals. However, a significant increase in the incidence of MNPCE in the bone marrow cell of the cyclophosphamide-treated mice was found. Moreover, in the groups treated with the total extract of A. leiocarpus at different doses plus cyclophosphamide, there was a significant decrease p < 0.0001 in MNPCEs compared to the positive controls, in a dose-dependent manner. Conclusion. This first finding reports that the extract of A. leiocarpus was neither genotoxic nor cytotoxic. However, it shows a protective effect against the genotoxicity and cytotoxicity induced by cyclophosphamide.

Study of the toxicological characteristics of plant extracts for use in sports nutrition products

В настоящее время имеются свидетельства, что ряд компонентов, образующихся в растениях, произрастающих в экстремальных условиях в ряде труднодоступных районов Южной Америки, Азии и Африки, в результате их успешного приспособления к особым условиям выживания, обладают уникальными физиологическими свойствами и способны оказывать мощное положительное воздействие на физическое и умственное здоровье людей. Вместе с этим точный состав этих растительных объектов и механизмы их воздействия на организм людей до настоящего времени изучены недостаточно. Это делает необходимым подробное исследование их токсикологических характеристик с целью получения экспериментальных доказательств возможности их безопасного применения в составе продуктов питания. Целью работы было изучение токсикологических характеристик новых перспективных компонентов спортивного питания - экстрактов маки перуанской, дамианы и L -теанина. Выявление биологической активности этих продуктов проводилось на классических модельных объектах, которые в настоящее время широко используются в подобных исследованиях во многих странах мира: на крови рыб данио ( Danio reriо ) и бактериях аэромонас ( Aeromonas gidrоphila ). Исследования, проведенные в направлениях выяснения влияния исследуемых продуктов на генотоксичность в эритроцитах рыб данио (микроядерный тест), на апоптоз (запрограммированную смену клеток) и эритропоэз (процесс кроветворения), показали, что ни один из исследованных препаратов не обладает генотоксичностью и не вызывает мутаций на молекулярном уровне. Было также обнаружено, что экстракты маки перуанской и дамианы подавляют рост условно патогенных бактерий человека, но не проявляют сильного токсического воздействия на микробиоту кишечника по сравнению со стандартным антибиотиком. Currently, there is evidence that a number of components formed in plants growing under extreme conditions in a number of hard-to-reach regions of South America, Asia and Africa, as a result of their successful adaptation to special conditions of survival, have unique physiological properties and are capable of exerting a powerful positive effect on physical and mental health of people. At the same time, the exact composition of these plant objects and the mechanisms of their effect on the human organism have not been sufficiently studied to date. This necessitates a detailed study of their toxicological characteristics in order to obtain experimental evidence of the possibility of their safe use in nutrition. The aim of the work was to study the toxicological characteristics of new promising components of sports nutrition - extracts of Maca Peruvian, Damiana and L -theanine. The investigation of the biological activity of these products was carried out on classical model objects, which are now widely used in similar studies in many countries of the world: on the blood of zebrafish ( Danio rerio ) and aeromonas bacteria ( Aeromonas gidrophila ). Studies carried out in the direction of elucidating the effect of the investigated products on genotoxicity in erythrocytes of zebrafish (micronucleus test), on apoptosis (programmed cell change) and erythropoiesis (hematopoiesis process) have shown that none of the investigated products has genotoxicity and does not cause mutations in molecular level. It was also found that extracts of Maca Peruvian and Damiana inhibit the growth of opportunistic human bacteria, but do not show a strong toxic effect on the intestinal microbiota compared to a standard antibiotic.