Genotoxic Effects Of Cadmium Research Articles

Genotoxic Effect of Cadmium and Zinc in the Peripheral Erythrocytes of Prussian Carp (Carassius Gibelio B.).

Cadmium and zinc are often found in aquatic environment and may accumulate in living organisms. The aim of this study was to evaluate the genotoxic effect of Cd, Zn, and their binary mixture on the peripheral blood erythrocytes of Prussian carp (Carassius gibelio B.). The fish were exposed to 4.0 mg/L Cd, 4.0 mg/L Zn or a mixture of 4.0 mg/L Cd and 4.0 mg/L Zn for a period of 14, 21 or 28 days. Genotoxic effects were investigated in peripheral blood cells using the comet assay and the erythrocyte micronucleus assay. The results demonstrated that the frequencies of micronuclei (MN) and both nuclear and cellular abnormalities in erythrocytes were significantly higher in all exposure groups as compared to the control group. The fish exposed to the mixture of Cd and Zn presented the highest frequency of MN. Furthermore, there was a decrease in the frequency of MN and an increase in the occurrence of DNA integrity defects (DNA damage) with longer time of exposure to the metals studied. Erythrocyte micronucleus and comet assays confirmed the genotoxicity of Cd and Zn. The results of the tests applied (which showed considerable variability) suggest the involvement of various toxicity mechanisms. Therefore, an integrative and comprehensive approach, using a set of assays for toxicity profile determination, should be adopted during ecotoxicological studies and environmental risk assessment pertaining to these elements.

Glial cytotoxicity of low doses of cadmium as a model of heavy metal pollution influence on vertebrates

Cadmium is a common transition metal that entails an extremely wide range of toxic effects in humans and animals. The cytotoxicity of cadmium ions and its compounds is due to various genotoxic effects, including both DNA damage and chromosomal aberrations. Some bone diseases, kidney and digestive system diseases are determined as pathologies that are closely associated with cadmium intoxication. In addition, cadmium is included in the list of carcinogens because of its ability to initiate the development of tumors of several forms of cancer under conditions of chronic or acute intoxication. Despite many studies of the effects of cadmium in animal models and cohorts of patients, in which cadmium effects has occurred, its molecular mechanisms of action are not fully understood. The genotoxic effects of cadmium and the induction of programmed cell death have attracted the attention of researchers in the last decade. In recent years, the results obtained for in vivo and in vitro experimental models have shown extremely high cytotoxicity of sublethal concentrations of cadmium and its compounds in various tissues. One of the most studied causes of cadmium cytotoxicity is the development of oxidative stress and associated oxidative damage to macromolecules of lipids, proteins and nucleic acids. Brain cells are most sensitive to oxidative damage and can be a critical target of cadmium cytotoxicity. Thus, oxidative damage caused by cadmium can initiate genotoxicity, programmed cell death and inhibit their viability in the human and animal brains. To test our hypothesis, cadmium cytotoxicity was assessed in vivo in U251 glioma cells through viability determinants and markers of oxidative stress and apoptosis. The result of the cell viability analysis showed the dose-dependent action of cadmium chloride in glioma cells, as well as the generation of oxidative stress (p <0.05). Calculated for 48 hours of exposure, the LD50 was 3.1 μg×ml-1. The rates of apoptotic death of glioma cells also progressively increased depending on the dose of cadmium ions. A high correlation between cadmium concentration and apoptotic response (p <0.01) was found for cells exposed to 3–4 μg×ml-1 cadmium chloride. Moreover, a significant correlation was found between oxidative stress (lipid peroxidation) and induction of apoptosis. The results indicate a strong relationship between the generation of oxidative damage by macromolecules and the initiation of programmed cell death in glial cells under conditions of low doses of cadmium chloride. The presented results show that cadmium ions can induce oxidative damage in brain cells and inhibit their viability through the induction of programmed death. Such effects of cadmium intoxication can be considered as a model of the impact of heavy metal pollution on vertebrates.

Genotoxic effect of Pb and Cd on in vitro cultures of Sphagnum palustre: An evaluation by ISSR markers

In the present work, the genotoxic effect of cadmium and lead supplied in a laboratory trial, was investigated for the first time in the moss Sphagnum palustre, by ISSR molecular markers. A total of 169 reproducible bands were obtained with 12 primers, ten of which gave polymorphisms (i.e., appearance/disappearance of bands), indicating a clear genotoxic effect induced by the metals. Both metals induced a decrease of the genome template stability in a dose dependent manner. At concentration >10−5 Cd also induced a general toxic effect in S. palustre, leading to chlorophyll degradation and moss death. Moreover, we followed the fate of supplied heavy metals into the moss tissue by SEM-EDX to see if they entered the cells. SEM-EDX observations on moss cultures treated with equimolar concentrations of the two metals showed that most Pb precipitated in form of particles on moss surface, while Cd did not aggregate in particles and was not found on moss surface. In light of these findings, we concluded that probably Pb induced a genotoxic effect at lower intracellular concentrations than Cd.

Detection of genotoxic effect of cadmium on tomato (Lycopersicon esculentum) using RAPD markers

Tomato (Lycopersicon esculentum) seeds were germinated in Peteri dishes for 7-10 days. Seedlings were transferred to pots and grown in greenhouse for two weeks. Cadmium treatments were applied with water with concentrations 0.0, 10.0, 50.0, 70.0 and 100.0 µM. Genomic DNA from tomato leaves was used for RAPD-PCR analysis using four random primers (OPB05, OPB07, OPB08 and OPB09). RAPD profiles showed great variations in banding patterns, particularly in intensity, number, thickness and mobility of the generated PCR fragments. Appearance of new PCR products may be occurred because some oligonucleotide priming sites could become accessible to primers due to changes in DNA sequence. Disappearance of some DNA fragments might be due to structural rearrangement in DNA caused by different types of DNA damage after exposure to cadmium pollution which can change the number of binding sites of Taq polymerase. Structural rearrangement may also results in an increase in the intensity and thickness of some DNA fragments. The change in the mobility of some DNA fragments might be due to Single Strand Conformational Polymorphism (SSCP) that reveals differences in electrophoretic mobility between normal and mutant single strands of DNA. These results indicated that RAPD analysis could be used as a useful biomarker assay for detection of genotoxic effects of cadmium. Cadmium pollution can cause genotoxic effects and tomato plants can be used as a bioindicator for evaluation of environmental pollution by heavy metals.

Genotoxic effects of cadmium and influence on fitness components of Lymantria dispar caterpillars

The current study extends our previous findings concerning the sensitivity of Lymantria dispar larvae to cadmium in light of ecotoxicological risk assessment. Here we report the results of the comet assay performed for the first time on this species. We examined the chronic effects of two cadmium concentrations (50 and 100 μg Cd/g dry food) on DNA integrity and haemocyte viability, as well as on fitness-related traits (larval mass and development duration parameters). All parameters were assessed individually and then used to calculate the integrated biomarker response (IBR) index. Egg-masses of L. dispar were collected from two locations in Serbia - the uncontaminated Homolje mountains and a metal-polluted area near Bor copper mines, smelter and refinery. Distinctive patterns in the response of these populations to cadmium exposure were noticed. In haemocytes of larvae from the pollution-free location both cadmium treatments increased the level of DNA damage, although in a similar range. Haemocyte viability and larval mass were reduced, while duration of the fourth instar and total development time were prolonged in a concentration-dependent manner. Cadmium tolerance was noticeable in the population from the metal-contaminated site at all organizational levels. Nevertheless, haemocyte viability in that population was reduced by the stronger treatment. Haemocyte viability was recognized as a promising biomarker due to the evident response of both populations to dietary cadmium. Genotoxicity, fitness-related traits and the IBR index could be used for biomonitoring of sensitive populations not previously exposed to metals.

Genotoxic effects of cadmium in human head and neck cell line SQ20B.

As cadmium may be involved in the etiology of head and neck cancers, we investigated in the present work, the cytotoxic and genotoxic effects of Cd on human larynx cells. SQ20B cells were exposed to 25 and 50μM Cd for 48 and 72h. Results showed a dose-dependent decrease in cell viability, especially after 48h, associated with mitochondria alterations as showed by transmission electronic microscopy. Surprisingly, the flow cytometry shows that the cells treated with Cd have a normal proliferative cycle like the untreated cell especially in G1 or G2 phase of cell cycle. DNA damages were investigated by comet assay and immunofluorescence for gamma layer of the H2AX (g-H2AX) foci formation. Results show a strong induction of DNA double-strand breaks after Cd exposure. Overall, our results demonstrate the cytotoxicity and genotoxicity of Cd in human larynx cells and support the view that Cd could be an etiologic factor of head and neck cancers.

The combined toxic and genotoxic effects of Cd and As to plant bioindicator Trifolium repens L.

This study was undertaken to investigate combined toxic and genotoxic effects of cadmium (Cd) and arsenic (As) on white clover, a pollutant sensitive plant frequently used as environmental bioindicator. Plants were exposed to soil spiked with increasing concentrations of cadmium sulfate (20, 40 and 60 mg Kg−1) or sodium arsenite (5, 10 and 20 mg Kg−1) as well as with their combinations. Metal(loid) bioavailability was assessed after soil contamination, whereas plant growth, metal(loid) concentration in plant organs and DNA damage were measured at the end of plant exposition. Results showed that individual and joint toxicity and genotoxicity were related to the concentration of Cd and As measured in plant organs, and that As concentration was the most relevant variable. Joint effects on plant growth were additive or synergistic, whereas joint genotoxic effects were additive or antagonistic. The interaction between Cd and As occurred at both soil and plant level. In soil the presence of As limited the bioavailability of Cd, whereas the presence of Cd increased the bioavailability of As. Nevertheless only As biovailability determined the amount of As absorbed by plants. The amount of Cd absorbed by plant was not linearly correlated with the fraction of bioavailable Cd in soil suggesting the involvement of additional factors, such as plant uptake mechanisms. These results reveal that the simultaneous presence in soil of Cd and As, although producing an additive or synergistic toxic effect on Trifolium repens L. growth, generates a lower DNA damage.

The Different Responses of Some Adult Albino Rats’ Body Organs to the Genotoxic Effect of Cadmium

The Different Responses of Some Adult Albino Rats’ Body Organs to the Genotoxic Effect of Cadmium

Green Algae Can Modulate the Genotoxic Action of Cadmium in Human Lymphocytes

ABSTRACTIn this paper we aimed to evaluate the cytotoxic, genotoxic and modulating potential of algal supernatants isolated from untreated cell suspension—SN(-), from algal cell suspension treated with CdCl2 10−6, 10−5 and 10−4 mol/l (SN), and from untreated algal cell suspension supplemented with various concentrations of CdCl2—10−6, 10−5 and 10−4 mol/l—SN(+). All data were compared with the cytotoxic and genotoxic effects of single cadmium treatment.Direct mutagenic assay and Cell-free medium/Lymphocytes assay were performed. Cell-wall-less strain cw15 of Chlamydomonas reinhardtii was grown for further procedures. Human lymphocytes were used as a valuable test system. The cytotoxic potential was evaluated on the basis of mitotic index (MI). The genotoxic potential was analysed on the basis of chromatid aberrations (CA).The data demonstrated that the supernatants SN(-), SN and SN(+) enhanced the mitotic activity of the cells comparing with single cadmium treatment. The most pronounced decreasing of the level of chromosome disturbances compared with the single cadmium treatment was seen only in the samples incubated with SN from algal suspension treated with the highest concentration of CdCl2—10−4 mol/l.Promising experimental evidence was provided that the cytotoxic and genotoxic effect of cadmium could be modulated as a result of the living activity of Chlamydomonas reinhardtii. The mode of action depends on the concentrations and the type of supernatants.

Cadmium-induced genotoxicity in zebrafish at environmentally relevant doses

Genotoxic effects of cadmium on zebra fish Danio rerio have been assessed by random amplified polymorphic DNA and real time PCR, followed by a comparison of the melting temperature patterns between each amplification reaction. Fish were exposed to two concentrations of cadmium chloride dissolved in the medium (1.9±0.6 μg Cd l −1, C 1; 9.6±2.9 μg Cd l −1, C 2) for 21 days. A discriminative RAPD probe, OPB11, was first selected producing differential band patterns between control and metal-exposed genomic DNAs. RAPD-PCR showed an increase in the relative hybridization efficiency of OPB11 on the genomic DNAs coming from fish exposed to both Cd concentrations as compared to the control condition. In addition, the RAPD-PCR melting temperature patterns showed that with the OPB11 probe, the frequency of PCR products whose fusion temperature belongs to the [86–87 °C] interval decreased with Cd contamination, whereas an increase of frequency for the [78–80 °C] and [85–86 °C] intervals was correlated with Cd exposure.

Cadmium‐ and copper‐induced DNA damage in Pisum sativum roots and leaves as determined by the Comet assay

Alkaline single‐cell gel electrophoresis (Comet assay) enables sensitive detection of DNA damage in eukaryotic cells induced by genotoxic agents. The present work aims to study the genotoxic effects of cadmium or copper in pea (Pisum sativum L.) roots and leaves. We performed the Comet assay on cells exposed to cadmium chloride (0.35 mg kg−1, 0.7 mg kg−1, 1.4 mg kg−1, 7.0 mg kg−1) or copper chloride (35 mg kg−1, 70 mg kg−1, 140 mg kg−1, 700 mg kg−1) for 30 days. The tail moment in migrated DNA was regarded as an indication of DNA damage. The genotoxic test revealed a pronounced DNA alteration in roots of plants exposed to cadmium, and less in those exposed to copper. However, DNA damage in leaves was only significant in plants treated with the higher cadmium concentrations. Analysis of cadmium accumulation showed that roots accumulated almost 50‐fold more cadmium than above‐ground parts of the plant. This may explain the low genotoxic effects observed in leaves of cadmium‐treated plants.

Genotoxicity of cadmium in marine diatom Chaetoceros tenuissimus using the alkaline Comet assay

Genotoxic effects of cadmium on phytoplankton Chaetoceros tenuissimus have been evaluated using DNA damage by Comet assay. Cadmium concentrations ranging from 2.4 to 10 mg/l were used to evaluate the effects. Results showed that as the concentration of Cd increased growth of the diatom decreased. Alkaline single-cell gel electrophoresis (Comet assay) method, which is highly sensitive in detection of DNA damage in eukaryotic cells, was used to observe genomic changes in marine diatom cells. DNA damage was measured as percent number of comets and normal cells. 65% cells were found to be damaged at 10 mg/l concentration of Cd as compared to 23% in 2.4 mg/l and only 5% in controls. More than 50% apoptotic cells were observed on 8th day at 10 mg/l and 12th day at 7.5 mg/l concentrations. At lower Cd concentrations (4.5 mg/l and below) the damage was below 30% till the last day. This suggested that higher Cd levels have early damaging effects on cell nuclear material and that % injury increases with advancement of exposure period. One advantage of use of C. tenuissimus is the ease with which it can be cultured in a defined medium. C. tenuissimus diatom can be used as an in vivo model for ecogenotoxicity assessment using the Comet assay.

Alleviation of free radical mediated oxidative and genotoxic effects of cadmium by farnesol in Swiss albino mice

Farnesol is an isoprenoid found in essential oils of ambrette seeds, citronella and in various aromatic plants. Exposure to cadmium from various sources affects the renal system adversely and Cd is an established genotoxic agent. In the present study, we evaluated the antigenotoxic and antioxidant efficacy of farnesol against cadmium chloride (CdCl2)-induced renal oxidative stress and genotoxicity in Swiss albino mice. Single, intraperitoneal doses of CdCl2(5 mg/kg body weight) for 24 h resulted in a significant (P < 0.001) increase in chromosomal aberration and micronuclei formation. The oral administration of farnesol at two doses (1% and 2% per kg body weight) for seven consecutive days showed significant (P < 0.05) suppression of the genotoxic effects of CdCl2 in the modulator groups. To study the mechanism by which farnesol exerts its antigenotoxic potential, enzymes involved in metabolism and detoxification were estimated. CdCl2 intoxication adversely affected the renal antioxidant armory and increased TBARS formation and xanthine oxidase levels significantly (P < 0.001). Farnesol showed a significant (P < 0.001) recovery in antioxidant status viz, GSH content (and its dependent enzymes) and catalase activity. Farnesol pretreatment in CdCl2-intoxicated mice showed marked (P < 0.001) suppression of TBARS' formation and XO activity. Our results support the conclusion that the anticlastogenic effect of farnesol could be due to restoration of antioxidants and inhibition of oxidative damage.

Evaluation of genotoxic damage of cadmium chloride in peripheral blood of suckling Wistar rats.

The aim of this study was to evaluate possible genotoxic damage of cadmium chloride exposure in suckling rats by means of the comet assay and the in vivo micronucleus test of rat blood lymphocytes, because no information is available on the genotoxic effect of cadmium in rats at this early age. Pups were receiving cadmium (as CdCl(2).H(2)O) orally in fractions of 0.5 mg for 9 days, totalling 4.5 mg Cd kg(-1) body wt, or were given a single subcutaneous injection of 0.5 mg Cd kg(-1) body wt. Some pups in both exposed groups were receiving calcium supplement (CaHPO(4).2H(2)O) in feed to reduce the body load of cadmium. Control pups did not receive either cadmium or calcium supplement. Cadmium in the carcass and organs was measured by atomic absorption spectrometry. The results showed that the cadmium body burden was significantly lower when the animals were receiving calcium supplements along with oral cadmium. The results of the micronucleus and comet assays showed significant differences between the control and exposed groups, regardless of the route of cadmium administration. The only statistically significant difference between the two exposed groups (oral cadmium and oral cadmium + calcium supplements) was in the number of micronuclei. The results of the comet assay showed that tail length differed statistically only between the control and all exposed groups, regardless of the route of cadmium administration. It can be concluded that the applied cadmium doses caused detectable genome damage but it was lower in calcium-treated pups receiving cadmium orally.

Genotoxic effect of cadmium is associated with apoptotic changes in tobacco cells

SUMMARYThis work explores the influence of cadmium on a suspension cell culture of Nicotiana tabacum (TBY‐2) by examining cell morphology, viability and DNA integrity. Changes in these parameters were strikingly dependent on concentration of cadmium in the culture medium: a concentration of 50–100 mmol m−3 CdSO4 induced apoptotic changes including DNA fragmentation into oligonucleosomal units, while 1 mol m−3 Cd2+ showed strong cytotoxicity, but no fragmentation of DNA. Low cadmium concentrations (below 10 mmol m−3) affected neither cell viability nor DNA integrity. A detailed kinetic study showed a significant delay in the onset of apoptosis after the application of high concentrations of cadmium. From days 0–3 after the application of 50 mmol m−3 CdSO4, the morphology of the cells, their viability and growth were indistinguishable between control and treated cells, and ‘domain’ DNA fragmentation into 50–200 kb fragments was observed at the DNA level. After this (days 4–7), there was a characteristic and rapid decrease in cell viability, distinct changes in cell morphology and oligonucleosomal fragmentation. The results suggest that chronic exposure of plant cells to cadmium can trigger programmed cell death.