Evaluation and quantification of genotoxicity of urban waters by using Vicia faba bioassays

The ecotoxic effect of pollutants in the water environment has become a hot topic of research. Because of the genotoxicity describing the composite deleterious action of the toxicants in water, the Vicia faba root-tip cell micronucleus (MN) test was applied for genotoxicity assessment of organic and inorganic substances in four rivers, two lakes and effluent flows from two wastewater treatment plants in Xi’an, China. Results showed that the lowest MN frequency was between 2.0‰ and 3.0‰. However, the highest MN frequency of the water samples was as high as 30.0–34.0‰. The results well reflected that the seriously polluted river with high chemical oxygen demand and total organic carbon showed high genotoxicity, and also indicated the genotoxic impact of urban discharge on the surface waters and the ability of urban water bodies to absorb the impact. In addition, there were significant correlations between the genotoxicity expressed by MN rate and Cr⁶⁺ (5–80?mg/L) and phenol concentrations (2–40?mg/L). The genotoxicity of the urban water could be quantified as equivalent Cr⁶⁺ or phenol concentrations and which might be used as a set of quantitative indicators of the water genotoxicity.     

Induction of micronuclei in tadpoles of Odontophrynus americanus (Amphibia: Leptodactylidae) by the pyrethroid insecticide cypermethrin

Cypermethrin (CY) is an active cyano pyrethroid effective against a wide range of pests encountered in agriculture and forestry. Although CY is not mutagenic in in vitro assays for gene mutation, in vivo assays showed conflicting results. In vivo genotoxicity of the synthetic pyrethroid CY in erythrocytes of Odontophrynus americanus tadpoles was examined. The frequency of micronuclei (MN) was recorded in blood smears obtained from tadpoles exposed in vivo to four different nominal concentrations 5, 10, 20 or 40?µg?L^?1 of the compound and fixed at two sampling times 48 and 96?h. As a positive control larvae were exposed to 40?mg?L^?1 of cyclophosphamide (CP). Tadpoles exposed to all CY treatments showed a significant increase in single small MN compared to the negative control group after 48?h and at 5 and 10?µg?L^?1 of CY at 96?h. Results obtained here demonstrated the genotoxic effects of the commercial formulation CY in the anuran larvae analyzed. Thus, data suggest that measurements of MN and other erythrocytes morphological aberrations performed in circulating blood samples of O. americanus tadpoles is a method for detecting cytogenetic damage in other native species.     

Cadmium accumulation by green algae Cladophora glomerata (L.) Kutz. (Chlorophyta) in presence of Nile tilapia Oreochromis niloticus (L.)

Accumulation of metals by aquatic organisms is mostly affected by other biological components in environments. In this study, cadmium (Cd) accumulation in green algae, Cladophora glomerata (L.) Kutz., exposed to 0.1 and 1.0 mg L^?1 of Cd for 15 and 30 days was examined in laboratory conditions in the presence of Nile tilapia Oreochromis niloticus (L.). The green algae C. glomerata accumulated Cd concentrations as 690 ± 70 and 3430 ± 470 mg kg^?1 on day 15, and 1130 ± 180 and 6830 ± 1540 mg kg^?1 on day 30. There were significant increases (p < 0.05) in metal accumulation by green algae as the exposure time and metal concentration increased. The results also indicated that the presence of Nile tilapia in the medium led to a significant Cd accumulation in the green algae compared to control (p < 0.05).     

Genotoxic effects of two different classes of insecticide in developing chick embryos

Pesticides provide considerable protection against pest population; however, rampant accumulation of these chemicals into varied habitats across the globe necessitates the need for a careful screening of each chemical due to toxic manifestations. In the current study, the genotoxic potential of two different classes of commercial insecticides – chlorpyrifos and cypermethrin combination and Spinosad, a naturalyte were compared. Rhode Island Red chick embryos were exposed to different doses of either of these insecticides individually, by in ovo treatment. Genotoxicity was then evaluated through micronucleus (MN) test and Comet assay. The combination insecticide exposure at low doses of 0.05 and 0.1 μg/egg induced DNA damage as evidenced by an increased tail moment in the Comet assay. Further, the presence of micronucleated erythrocytes and also various abnormal cells including dacryocytes, microcytes, erythroplastids, squashed/notched nuclei, and spindle-shaped erythrocytes in the blood smear consolidates indicate the presence of insecticide-induced genotoxicity. Spinosad, however, was found only mildly genotoxic but at a high dose of 1.5 mg/egg. The results indicate that usage of naturalyte insecticide may be a better option to minimize the harmful effects of chemical insecticides.     

Lysosomal,genetic and chromosomal damage in haemocytes of the freshwater bivalve (Unio pictorum) exposed to polluted sediments from the River Cecina (Italy)

The response to chemical disturbance of an Italian river (Cecina River) characterised by high levels of trace metals was assessed in the sentinel species freshwater painter’s mussel (Unio pictorum). We exposed U. pictorum in the laboratory to metal, metalloid and other trace element polluted river sediments in order to evaluate genetic and cellular biomarkers. Bivalves exposed to sediments taken from all the impacted sites of the river basin exhibited a significant impairment at the DNA, chromosomal and lysosomal levels. The lysosomal membrane stability of circulating haemocytes, assessed by the Neutral Red Retention Time, was found to be higher in specimens from Lake Maggiore and Berignone, while exhibited a significant decrease in specimens exposed to Ponteginori sediments. An increase of DNA migration in specimens exposed to the two contaminated sites in comparison with controls was found. Moreover, higher micronucleated haemocytes frequencies were observed in bivalves exposed to sediments collected at Possera 1 site. This study suggested that biomarkers evaluating cellular impairment, genetic and chromosomal damages, are highly sensitive and suitably applied to laboratory experiments. The painter’s mussel was confirmed to be an appropriate sentinel species for quality assessment in metal-contaminated freshwater environments.     

Nile water pollution and technical efficiency of crop production in Egypt: an assessment using spatial and non-parametric modelling

Agriculture is considered one of the vital activities in Egypt; it consumes about 83 % of the Egyptian Nile water quota. This activity is becoming negatively affected by water pollution causing negative repercussion on land productivity and subsequently food security. This paper assesses the water quality for agriculture along the mainstream of the Nile in Egypt through spatial distributions modelling of total dissolved solids (TDS), using spatial statistical analysis. The study’s sample frame consists of 78 sampling points collected in February 2008 and located on the Nile mainstream and its two branches, Rosetta and Damietta. Exploratory spatial data analysis is carried out on the TDS, followed by plotting and modelling the experimental semi-variogram. Then, cross validation is executed in order to determine the best fitting model. Finally, surface mapping is generated by performing spatial interpolation, using kriging technique. The generated surface map shows that the TDS levels increase from Upper to Lower Egypt, exceeds the standard limit in Beni-Suef and Rosetta branch. In fact, high levels of TDS are known to have a negative effect on Egyptian agriculture through harmfully affecting the soil and consequently the crop yields. Therefore, an analysis of the effect of water pollution on technical efficiency of crop production is conducted using a non-parametric mathematical programming approach to frontier estimation. The results of this estimation indicated that the TDS is overutilized in all governorates except Aswan.     

Cytogenotoxicity and histopathological assessment of Lekki Lagoon and Ogun River in Synodontis clarias (Linnaeus, 1758)

The cytogenotoxicity and histopathological alterations induced by xenobiotics in Lekki Lagoon and Ogun River on Synodontis clarias were investigated. Fish from these water bodies and a fish farm (control) were examined for micronucleated, binucleated, and immature erythrocytes in both gill and peripheral blood. Also gill, liver, kidney, and ovary were processed for histopathology using hematoxylin-eosin staining. Concentrations of cadmium, zinc, lead and copper in the water were determined. There was significant (p < 0.05) increase in micronucleated, binucleated, and immature erythrocytes in both gill and peripheral blood of S. clarias from the lagoon and river compared to the reference site. Loss and disorganization of the primary and secondary lamellae, multifocal degeneration, hemorrhages, cellular infiltration, congestions, vacuolations, atresia, and necrosis were common lesions in the examined tissues of fish from the lagoon and river. Cd, Zn, Pb, and Cu in water samples from the lagoon and river were higher than the reference site. Xenobiotics in Lekki Lagoon and Ogun River, mostly metals, induced deoxyribonucleic acid (DNA) and pathological damage in S. clarias.     

Genotoxic assessment of anthracene and benzo [a] pyrene to milkfish Chanos chanos

Mutagenic and genotoxic effects of polycyclic aromatic hydrocarbons, anthracene and benzo [a] pyrene (BaP), in milkfish Chanos chanos were determined using micronucleus (MN) test and comet assay (CA). Distinct mean frequencies of nuclear abnormalities such as MNs; binucleated micronuclei, nuclear bud, and fragmented apoptotic cells were measured. Significant increase in DNA damage with five classes of damage level was observed and expressed in terms of arbitrary unit (AU). Mean frequencies of total nuclear abnormalities were 0.5?±?0.25 cells in control; 0.67?±?0.33 cells in solvent control; 70?±?9.60 cells in 0.176?mg?L^?1 anthracene, and 91.83?±?6.25 cells in 0.031?mg?L^?1 BaP. The greatest DNA damage of 170AU was observed in 0.176?mg?L^?1 anthracene-exposed group and 182AU was observed in 0.031?mg?L^?1 BaP-treated fish. This study confirmed that the CA and MN assays are useful tools in determining potential genotoxicity of water pollutants and might be appropriate as a part of monitoring program.     

Observations on metal concentrations in commercial landings of two species of tilapia (Oreochromis mossambicus and Oreochromis niloticus) from reservoirs in six river basins in Sri Lanka

Samples of the muscle of two species of tilapia (Oreochromis mossambicus and O. niloticus; 17–20 cm length) were obtained from at least one reservoir in each of the six river basins (Aruvi Aru, Kala Oya, Kirindi Oya, Ma Oya, Mahaweli, and Walawe Ganga catchments) in Sri Lanka. The metals Ca, Cu, Fe, K, Mg, Mn, Na, and Zn were consistently detected in the muscle tissue. Overall, there were few differences in the concentration of metals between the two species of fish, although there were also some statistically significant differences (p < 0.05) in the concentrations of some metals in fish obtained from some of the reservoirs. Aruvi Aru stands out as a river basin in which the two fish species have significantly lower concentration of metals when compared to other river basins. The concentration of the metals studied were below WHO and FSANZ guideline values for fish, suggesting that the consumption of the metals found in tilapia from these reservoirs poses little risk to human health.     

Dissolved Ions of Trace and Major Elements and in Suspended Sediments in the Nile, Egypt

Dissolved ions and suspended participates from twelve water samples from Sohag (different localities) and Aswan (behind and below the High Dam) areas were collected along the River Nile, upper Egypt, which is at present affected seriously by pressure of population, intense industrial and agricultural activities, and atmospheric fallout. Eighteen trace and nine major ions in solution and fourteen metals in suspension were analyzed to establish whether elements are of anthropogenic or natural origin. Results of this study show that the Nile water was contaminated with high concentrations of dissolved lead, and with copper, nickel and mercury to a lesser extent, suggesting that these contaminants are generated from anthropogenic activities. the other dissolved ions in the Nile are of natural origin because their metal concentrations fall within the range of the normal background and average world dissolved values for inland waters. Results show that the usual constituents of suspended particulates in the Nile seem to be some metals (Cu and Zn) adsorbed on iron and manganese oxide/hydroxide. the results also reveal that most trace and major metal particulates increase from south to north due to activities associated with the dense population living along the Nile banks. No significant differences were observed between the chemical composition of trace and major elements taken on the east and west river Nile banks.     

Assessment of micronuclei and chromosomal anomalies of five biocompatible materials in mice

In vivo genetic toxicology tests measure direct DNA damage or the formation of gene or chromosomal mutations, and are used to predict mutagenic and carcinogenic potential of compounds for regulatory purposes. These adverse genotoxic effects may be manifested in the form of gene mutations, structural chromosomal aberrations (CA), recombination, and numerical changes. The present investigation was carried to assess genotoxic effects of five different implantable biomaterials developed in different laborataries of Sree Chitra Tirunal Institute of Medical Sciences and Technology. All biomaterials were developed for clinical applications. CA and micronuclei (MN) studies are biomarkers of genotoxicity testing. Leachants from the extract of biomaterials are capable of inducing structural and numerical chromosomal changes. The studies were conducted in Swiss albino mice with the physiological saline extract of materials together with cyclophosphamide and physiological saline as positive and negative controls. Animals were administered intraperitoneally (ip) with a single injection of test, positive (cyclophosphamide), and negative (physiological saline) control and sacrificed after 24 or 48?h. Bone marrow cells were collected for CA and MN assays. Data showed that all five biomaterials did not significantly exert genotoxic effects. Hence, the study indicates that these biomaterials do not induce any chromosomal anomalies.     

Genotoxic Effect of Phenol on the Cells of Onion (Allium cepa) Roots

The roots of onion (Allium cepa) stand out for having cells with large size and small number of chromosomes.These characteristics make them useful in bioassays for the measurement of a variety of cytogenetic and morphological parameters , in which they can be used as toxicity indicators of the induction and formation of micronuclei and chromosomal aberrations.Based on this background , the potential genotoxic effect of phenol concentration on cells of A.ceparoots was investigated either in terms of induced aberrations or micronuclei formation.The results demonstrated that the higher the concentration of phenol , the higher the incidence of abnormalities , thus confirming the genotoxicity of this pollutant.     

Genotoxicity and mutagenicity of electronic waste leachates using animal bioassays

Human exposure to electronic waste (e-waste) is increasing, especially in developing nations. This has potential short and chronic adverse health risks. Information is currently scarce on potential genotoxicity of e-waste leachate. In this study, the potential mutagenic and genotoxic effects of leachates from e-waste dumpsites at two electronic markets in Lagos, Nigeria, were investigated. The assays utilized were: bone marrow micronucleus (MN) and chromosome aberration (CA), spermatogonia CA, sperm morphology, and sperm count in mice. Experiments were carried out at concentrations of 1%, 5%, 10%, 25%, and 50% (v/v; leachate: distilled water) of leachate samples. MN analysis showed a concentration-dependent induction of micronucleated polychromatic erythrocytes (MNPCE) across the treatment groups. In the CA tests (bone marrow and spermatogonia), there was concentration-dependent significant reduction in mitotic index and induction of different types of CA. Assessment of sperm shape showed a significant increase in sperm abnormalities with significant decrease in mean sperm count in treated groups. Heavy metals analyzed in the tested samples are believed to contribute significantly to the observed genetic damage. This indicates that e-waste contains potential genotoxic agents; and constitutes a genetic risk in exposed human population.     

A study on genotoxicity of textile dyeing industry effluents from Rajshahi,Bangladesh, by the Allium cepa test

Textile dye effluents are believed to be toxic as they might exert various harmful effects on living organisms including genotoxicity. These effluents are the main source of direct and continuous input of pollutants into the aquatic ecosystems. In this study, dye effluents from a local silk dyeing industry were analysed for their genotoxic potential by the Allium cepa genotoxicity test. The A. cepa test is characterised as a genotoxicity test where the roots of A. cepa are grown in different concentrations of the test material. The macroscopic results clearly showed that the toxicity of the dye effluents prompted A. cepa root growth inhibition, and this effect increased with higher concentrations of the effluents. At the cellular level, no dividing cells were found at higher concentrations such as 60%, 80% and 100% of the effluents. However, at a lower concentration of 20%, dividing cells were identified, although the mitotic index was much lower than that of the control. Microscopic analysis showed that the dye effluents induced chromosomal aberrations at significant levels. Taken together, these results revealed that the textile dyeing industry effluents are toxic to eukaryotic cells and these dyes have genotoxic properties that can potentially lead to cancer development and adverse health conditions.     

Evaluation of 3-hydroxy-benzo[a]pyrene levels in Nile tilapia (Oreochromis niloticus) after waterborne exposure to Benzo[a]pyrene

3-Hydroxy-benzo[a]pyrene (3-OH-BaP), a toxic compound with the ability to covalently bind with the macromolecules (proteins and DNA), is one of the major phenolic metabolites of benzo[a]pyrene (BaP). The purpose of this study was to evaluate the presence of 3-OH-BaP in the bile and plasma of Nile tilapia by HPLC with fluorescence detection, after waterborne exposure to BaP (10 and 100?µg?L^?1). Metabolites were detected in bile and plasma, and conjugates of 3-OH-BaP (glucuronide and/or sulphate conjugates) were the majority forms in both biological fluids, glucuronide 3-OH-BaP being the main metabolite in bile. Our data suggest that extrahepatic tissues as intestine or gill are important in BaP metabolism and need to be the considered sources of metabolites released into the blood. Although, low levels of 3-OH-BaP in toxic form (free form) were detected in plasma, one cannot exclude the possibility of circulating the levels leading to adverse effects.     

Hematological studies on some Nile fishes,Tilapia nilotica,Lates niloticus,and Labeo niloticus

Hemoglobin content of 3 Nile fishes — Tilapia nilotica Linnaeus, Lates niloticus Linnaeus and Labeo niloticus Forskal— were examined. Clear differences in hemoglobin and hematocrit values were observed, and seasonal variation in the hemoglobin content of Lates niloticus and Labeo niloticus were noticed. There is a correlation between hematocrit value and hemoglobin content of the 3 species examined. The chloride content is higher in the species with lower hemoglobin content.     

Histopathological alterations of Nile tilapia, Oreochromis niloticus in acute and subchronic alachlor exposure

Histopathological alterations in Nile tilapia, Oreochromis niloticus, aged 3 months and subjected to acute and subchronic alachlor exposure were studied by light microscopy LC50 values of alachlor for 24 hr 48 hr, 72 hr and 96 hr were 963.6, 563, 448, and 381.9 microg l(-1), respectively and the maximum acceptable toxicant concentration (MATC) was 350 microg l(-1). Fish were exposed to 381.9 (acute) and 35 microg l(-1) (sub-chronic) of alachlor for 24, 48, 72 and 96 hr and 90 days, respectively Gill lamellae and kidney tubules were the primary target organs for the acute toxic effect of alachlor while in the subchronic exposure, the toxic effect on the gills was less marked than that of the kidneys and liver Gill alterations included edema of the epithelial cell system, aneurisms with some ruptures, hypertrophy and hyperplasia of epithelial cells. The liver showed hydropic swelling of hepatocytes and vacuolation. Lipid vacuoles were observed in hepatocytes in the second and third month of subchronic exposure. The kidney showed hydropic swelling of tubular cells, lipid vacuole accumulation in many tubules, and nuclear pyknosis. The findings of this study could be used as a guideline forbiomonitoring programs on populations of Nile tilapia cultured near alachlor contaminated areas.     

Electrosensory prey detection in the African sharptooth catfish, Clarias gariepinus (Clariidae), of a weakly electric mormyrid fish, the bulldog (Marcusenius macrolepidotus)

During their seasonal runs in the Okavango and other freshwater bodies in southern Africa, the sharptooth catfish, Clarias gariepinus, specialises on the bulldog, Marcusenius macrolepidotus, as its main prey. We examined whether the catfish can locate bulldogs by detecting their pulse-type electric organ discharges (EODs). The electrosensory threshold for single-cycle, monopolar square-wave pulses was exceedingly low (down to 13 μV_p–p/cm for 4-ms pulses) in trained, food-rewarded sharptooth catfish (n=4), confirming the results of Lissmann and Machin who were, however, unable to identify a biological function. Other stimulus pulse waveforms (single-cycle, monopolar as well as bipolar sine-wave pulses) were also effective stimuli according to their spectral low-frequency energy contents (0– 30 Hz). Male bulldogs display an EOD pulse approximately 10× the duration of female EODs (≈0.5 ms). The C. gariepinus threshold for field-recorded playbacks of a male bulldog EOD (of long duration) was 103 μV_p–p/cm, whereas the brief female and juvenile EODs were not detected (using intensities of natural EODs). EODs of other mormyrids were detected when either monopolar or of long duration. Signal source amplitude increased linearly with standard length (SL) in bulldogs. Signal reach, as calculated from signal source amplitude and receiver sensitivity, is up to 150 cm for a large male bulldog (SL 27.5 cm), and 83 cm for a male that has just turned sexually mature (SL 12.6 cm). Therefore, most bulldogs eaten by catfish are probably male, in agreement with the size distribution of bulldogs found in catfish stomachs. These results suggest that sharptooth catfish rely heavily on their acute electrical sense during hunting, and an important function for electroreception in an African catfish has been identified. Received: 13 December 1999 / Revised: 20 April 2000 / Accepted: 20 May 2000     

利用微核试验和彗星电泳试验评价黄河兰州段水质致遗传毒性作用

利用微核和彗星电泳试验对黄河兰州段鲤鱼(Cyprinuscarpio)红细胞大小及其DNA损伤(微核、核异常、单链断裂和遇碱不稳定点、双链断裂)进行了研究,并以刘家峡水库为对照.结果显示,黄河兰州段采得的鲤鱼其红细胞微核率显著增加(P<0.05),DNA单链断裂和遇碱不稳定点损伤比例和程度加重,但两样点差异不显著(P>0.05),这表明兰州段黄河水质有明显的致遗传毒性作用.同时两种检测结果显示,不同的遗传毒性检测手段具有不同的检测终点和机制,建议在监测环境污染致遗传毒性作用时考虑检测方法的统一和标准化,从而得到更可靠、全面的结果.图4表3参15     

两种喂食方式下微囊藻毒素-LR在罗非鱼肝脏和肌肉中动态变化的比较

试验以尼罗罗非鱼作为受试动物,通过直接投喂未经处理的新鲜藻细胞和喂食相同量经超声波破碎过的藻细胞,比较了罗非鱼在摄食完整蓝藻细胞与摄食破碎的蓝藻细胞时微囊藻毒素-LR(MC-LR)在其体内的动态变化规律及代谢差异.结果表明,无论是喂食破碎蓝藻细胞还是未破碎的蓝藻细胞均可在罗非鱼肝脏和肌肉中检出MC-LR,细胞破碎组肝脏和肌肉的富集能力高于未破碎组.富集第3d,即可在肝脏和肌肉中检出不同程度的MC-LR,肝脏中MC-LR的含量显著高于肌肉,随着富集时间的延长,MC-LR的含量总体呈上升趋势,破碎组肝脏和肌肉分别在染毒的16d和11d达到最大值(分别为2.021μg·g-1和0.071μg·g-1);未破碎组则分别在染毒的24d和21d达到最大值(分别为1.856μg·g-1和0.036μg·g-1).整个富集阶段破碎组肝脏MC-LR的平均值为1.171μg·g-1,略高于未破碎组的1.029μg·g-1;破碎组肌肉含量的平均值0.051μg·g-1,高于未破碎组的0.029μg·g-1.将细胞未破碎组的罗非鱼进行释放阶段的试验.结果表明,罗非鱼的肝脏能快速清除MC-LR,释放13d时达到释放阶段的最低值(为0.241μg·g-1),与肝脏相比,肌肉对毒素的清除要缓慢得多.