Determination of micronuclei frequency in Danio rerio for assessing genotoxicity induced by propineb.

The aim of this study was to investigate the genotoxic effect of Propineb fungicide at different concentrations (0.167, 0.335 and 0.670 mg L-1) and different treatment times (24, 48, 72 and 96 h) on Danio rerio. At the end of the treatment periods, blood was collected from the fish with a heparin injector; smear preparations were prepared, fixed and stained. In the prepared preparations, the numbers of cells with MN and erythrocyte nucleus abnormalities were examined. It was found that propineb increased micronucleus formation at all treatment times and concentrations and induced the formation of erythrocytes with morphological abnormal nuclei such as segmented, kidney-shaped, notched, vacuolated nuclei and binucleated. The increase in micronucleus formation and the number of erythrocytes with abnormal nuclei were found to be concentration and treatment time-dependent. In conclusion, in this study, Danio rerio erythrocytes were used to evaluate the genotoxic effects of propineb fungicide on aquatic organisms, which have an important place in environmental risk assessment criteria. Since fungicides used in agricultural control such as propineb may have the potential to be genotoxic to aquatic organisms, the results of toxicity tests should be taken into consideration in the selection and use of concentrations of these chemicals.

Investigation of genotoxic effects of rhododendron honey using three mammalian bioassays in vivo.

Rhododendron honey (RH) is obtained from the rhododendron plants are grown in many regions around the world, causes poisoning in humans due to the grayanotoxin (GTX) compound in its structure. It is used by the public as a therapeutic for some diseases. It was aimed to study the genotoxic and cytotoxic effects of RH in mouse bone-marrow and sperm cells by using three mammalian bioassays. 25, 50 and 75 mg kg-1 concentrations of RH given to male mice via gavage for 24 and 48 h treatment periods and its active ingredient Grayanatoxin (GTX-III) 0.01 mg kg-1 by i.p. injection. Chromosome aberrations (CA), polychromatic erythrocytes (PCE)/normochromatic erythrocytes (NCE), micronucleated polychromatic erythrocytes (MNPCE) and sperm abnormalities were investigated. The results demonstrated that all the tested concentrations of RH significantly induced total abnormal cell frequency including chromosomal breaks for two time periods. In the MN assay, 75 mg kg-1 RH and 0.01 mg kg-1 GTX-III significantly increased % MNPCE and significantly reduced PCE/NCE ratios after 24 and 48 h treatments on mice demonstrating potential genotoxic and cytotoxic effect. Although there was a concentration-related increase in the percentage of total sperm abnormalities, this increase was not statistically significant compared to control. As a result, microscopic genotoxicity and cytotoxicity marker tests showed that RH and its active ingredient GTX-III have potential genotoxic and cytotoxic effect on mice bone marrow cells. It is understood that RH that is used to treat some diseases by public, should be handled carefully and used in a controlled manner.HighlightsChromosome aberration, micronucleus and sperm morphology assays are recommended as reliable biological indicators.RH and its active ingredient GTX-III have potential genotoxic and cytotoxic effect on mice bone marrow cells.Significant changes were observed upon the treatment of 75 mg kg-1 MH for MN assay.

The effect of rhododendron honey on mice liver tissue / Efecto de la miel de rododendro en el tejido hepático de ratón

Rhododendron honey, made by bees from rhododendron pollen, contains a toxic substance called grayanotoxin. Depending on the dose, the poisonous honey can result in serious effects such as cardiac arrhythmia, fibrillation, and myocardial infarction. The purpose of this study is to investigate the effects of the poisonous RH of the Black Sea Region on the liver. Male mice were divided into five groups of twelve mice each, two being the control groups (distilled water) and the others being the rhododendron honey (RH) groups (25, 50, and 75 mg/kg) and 0.01 mg/kg grayanotoxin (GTx) groups. Liver tissues were collected 24 and 48 h later. The sections were stained with hematoxylin, eosin and PAS, then the histopathological score was performed. Significant statistical differences were observed between the RH and control groups in terms of congestion, steatosis, sinusoid dilatation, and inflammation. The control group demonstrated a normal liver structure in the light microscopy, while the GTx-applied 24 h group exhibited expansions in the sinusoids and congestion. Higher levels of congestion, steatosis, and inflammatory cells were seen in the GTx-applied 48 h group. In the same group, giant cells consisting of many nuclei were observed in the sinusoids. The results of the 25 mg RH-applied groups were similar in 24 and 48 h, histopathological score levels were increased slightly, congestion and steatosis were prominent in the 48 h group. Dense steatosis was seen in the hepatocytes around the vena centralis in 50 mg/kg RH-applied 48 h group. Congestion, steatosis and an increase in inflammatory cells were observed in the hepatocytes in the 75 mg/kg RH-applied 24- and 48 h groups. PAS (+) stained hepatocytes were decreased in the RH- and GTx-applied groups. The toxic effects of the rhododendron honey were observed in the mice liver tissue with respect to dose and time.

La miel de rododendro, elaborada por las abejas a partir del polen de rododendro, contiene una sustancia tóxica llamada grayanotoxina. Dependiendo de la dosis, la miel venenosa puede resultar en efectos graves, tales como arritmia cardiaca, fibrilación e infarto de miocardio. El propósito de este estudio fue investigar los efectos en el hígado de la miel venenosa de rododendro de la región del Mar Negro. Se distribuyeron ratones machos en cinco grupos de doce ratones cada uno, dos grupos control (agua destilada) y los otros grupos se trataron con la miel de rododendro (MR) (25, 50 y 75 mg/kg) y con 0,01 mg/kg grayanotoxina (GTX). Los tejidos hepáticos se recogieron 24 y 48 h más tarde. Las secciones fueron teñidas con hematoxilina-eosina y PAS. A continuación, se realizó la puntuación histopatológica. No se observaron diferencias estadísticamente significativas entre MR y los grupos de control en términos de congestión, esteatosis, dilatación sinusoidal e inflamación. El grupo control demostró una estructura normal del hígado en el microscopio de luz, mientras que el grupo de las 24 horas de aplicación de GTX exhibió expansiones en los sinusoides y congestión. Mayores niveles de congestión, esteatosis y células inflamatorias se observaron en el grupo de 48-horas de aplicación de GTX. En el mismo grupo, se observaron células gigantes que consistían en la presencia de muchos núcleos en los sinusoides. Los resultados de los grupos con aplicación de 25 mg de RH fueron similares en los resultados de 24 y 48 h, los niveles de puntuación histopatológica aumentaron ligeramente, la congestión y la esteatosis fueron prominentes en el grupo de 48 h. Se observó esteatosis densa en los hepatocitos en toda la vena central en el grupo de aplicación de 50 mg/kg de RH, 48 h. La congestión, la esteatosis y un aumento en las células inflamatorias se observaron en los hepatocitos en el grupo de 75 mg/kg de MR de 24 h y los grupos de 48 h. Hepatocitos teñidos con PAS (+) disminuyeron en los grupos de GTX y MR. Se observaron los efectos tóxicos de la miel de rododendro en el tejido hepático de ratones con respecto a la dosis y el tiempo.

Abnormal sperm morphology in mouse germ cells after short-term exposures to acetamiprid, propineb, and their mixture.

Pesticides are one of the most potent environmental contaminants, which accumulate in biotic and abiotic components of ecosystems. Acetamiprid (Acm), a neonicotinoid insecticide, and Propineb (Pro), a dithiocarbamate fungicide, are widely used to control sucking insects and fungal infections on crops, respectively. The present study was undertaken to investigate the genotoxic effects of these compounds, individually and in mixtures, in mouse germ cells by using the sperm morphology assay. Mice were injected intraperitoneally with 0.625, 1.25, and 2.50 µg mL⁻¹ of Acm, 12.5, 25, and 50 µg mL⁻¹ of Pro, and their mixture at the same concentrations over 24 and 48 h. Acm did not significantly increase the percentage of abnormal sperm at any concentration. The frequency of abnormal sperm significantly increased after 24 and 48 h of exposure to 50 µg mL⁻¹ of Pro. The mixtures of 2.50 µg mL⁻¹ of Acm and 50 µg mL⁻¹ of Pro induced sperm abnormalities antagonistically both after 24 and 48 h of exposure. Results suggest that Acm was non-genotoxic for mouse germ cells, while Pro may have been a germ cell mutagen due to the observed increase in the frequency of sperm abnormalities. However, to gain better insight into the mutagenicity and DNA damaging potential of both of these pesticides, further studies at molecular level should be done.

Chromosome aberrations, micronucleus and sperm head abnormalities in mice treated with natamycin, [corrected] a food preservative.

Natamycin [corrected] is used as preservative in foods. The genotoxic effects of the food preservative natamycin [corrected] were evaluated using chromosome aberrations and micronucleus test in bone marrow cells and sperm head abnormality assays in mice. Blood samples were taken from mice and levels of total testosterone in serum were also determined. Natamycin [corrected] was intraperitoneally (ip) injected at 200, 400 and 800 mg/kg. Natamycin [corrected] did not induce chromosome aberrations but significantly increased the number of micronucleated polychromatic erythrocytes in bone marrow and sperm head abnormalities at all concentrations and treatment periods. It also decreased MI at all concentrations for 6, 12 and 24h treatment periods. Natamycin [corrected] decreased PCE/NCE ratio at all concentrations for 48h in female mice, for 24 and 48h treatment periods in male mice. At the 800 mg/kg concentration, natamycin [corrected] decreased PCE/NCE ratio for 24 and 72h in female mice. A dose dependent increase was observed in the percentage of sperm head abnormalities. The levels of serum testosterone decreased dose-dependently. The obtained results indicate that natamycin [corrected] is not clastogenic, but it is aneugenic in mice bone marrow and it is a potential germ cell mutagen in sperm cells.