A Chromatographic Approach for High-Precision Eu Isotope Analysis.

Eu isotopes are promising tracers across various scientific domains such as planetary, earth, and marine science, yet their high-precision analysis has been challenging due to the similar geochemical properties of rare earth elements (REEs). In this study, a novel two-column chromatographic approach was developed utilizing AG50W-X12 and TODGA resins to separate Eu effectively from matrix and interfering elements like Ba, Nd, Sm, and Gd, while ensuring high Eu yields (99.4 ± 0.4%, n = 19) and low blanks (<20 pg). The robustness of this method is evidenced by various rock types and different Eu loading masses. The efficient purification of Eu facilitated the establishment of a high-precision calibration technique with standard-sample bracketing (SSB) and internal normalization (Nd). When a Nu Plasma 1700 multicollector-inductively coupled plasma-mass spectrometry (MC-ICP-MS) instrument was employed, repeated purification and analysis of various Geological Reference Materials (GRMs) confirmed that the long-term external precision of δ153/151Eu is better than 0.04‰ (2 standard deviation (2SD)), which represents a 2-5-fold increase in precision compared to previously reported methods. Additionally, the high-precision Eu isotopic compositions of five GRMs, including basalts, andesite, syenite, and marine sediment, were measured. The high-precision Eu isotope techniques presented herein open up new avenues for Eu isotope geochemistry.

Different impacts of natural and anthropogenic factors on dissolved organic matter chemistry in coastal rivers: Implications for water management.

The chemical composition of dissolved organic matter (DOM) exerts significant influence on aquatic energy dynamics, pollutant transportation, and carbon storage, thereby playing pivotal roles in the local water quality and regional-global biogeochemical cycling. However, the effects of natural climate change and local human activities on watershed characteristics and in-river processes have led to uncertainties regarding their contributions to DOM chemistry in coastal rivers, creating challenges for effective water management and the study of organic matter cycling. In this investigation, we employed a combination of stable isotopic analysis, optical techniques, and Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) to elucidate the sources, optical properties, and molecular composition of DOM in three South China coastal rivers. Our results suggest that terrestrial DOM entering the three rivers through natural or anthropogenic pathways is gradually diluted by in situ primary production as it moves downstream, ultimately being influenced by seawater intrusion near the estuary. Additionally, terrestrial processes influenced by temperature likely govern DOC concentration, while seawater intrusion promotes the natural production of S-containing organic compounds. In contrast, human-altered landcover significantly impacts DOM molecular composition. Increased water areas lead to the enrichment of lignins with high disinfection byproduct formation potential, and agricultural residue burning appears to be the dominant source of pyrogenic DOM in these coastal rivers. Our distinct results suggest that the development of specific water management plans that consider the combined effects of temperature, seawater intrusion, landcover changes, and agricultural practices will be essential to ensure sustainable water resource.

Metagenomics and Stable Isotopes Uncover the Augmented Sulfide-Driven Autotrophic Denitrification in a Seasonally Hypoxic, Sulfate-Abundant Reservoir.

The mechanism governing sulfur cycling in nitrate reduction within sulfate-rich reservoirs during seasonal hypoxic conditions remains poorly understood. This study employs nitrogen and oxygen isotope fractionation in nitrate, along with metagenomic sequencing to elucidate the intricacies of the coupled sulfur oxidation and nitrate reduction process in the water column. In the Aha reservoir, a typical seasonally stratified water body, we observed the coexistence of denitrification, bacterial sulfide oxidation, and bacterial sulfate reduction in hypoxic conditions. This is substantiated by the presence of abundant N/S-related genes (nosZ and aprAB/dsrAB) and fluctuations in N/S species. The lower 15εNO3/18εNO3 ratio (0.60) observed in this study, compared to heterotrophic denitrification, strongly supports the occurrence of sulfur-driven denitrification. Furthermore, we found a robust positive correlation between the metabolic potential of bacterial sulfide oxidation and denitrification (p < 0.05), emphasizing the role of sulfide produced via sulfate reduction in enhancing denitrification. Sulfide-driven denitrification relied on ∑S2- as the primary electron donor preferentially oxidized by denitrification. The pivotal genus, Sulfuritalea, emerged as a central player in both denitrification and sulfide oxidation processes in hypoxic water bodies. Our study provides compelling evidence that sulfides assume a critical role in regulating denitrification in hypoxic water within an ecosystem where their contribution to the overall nitrogen cycle was previously underestimated.

Calcium isotope fractionation during weathering of argillaceous carbonates in a humid climate.

The continental weathering is a key process that controls calcium (Ca) transportation from the continental crust to the waters. To elucidate the behavior of Ca isotopes during carbonate weathering, the concentrations and δ44/40Ca (relative to NIST SRM 915a) of bulk saprolites, exchangeable, acid-leachable and residual phases of a weathering profile developed on the marine carbonates, Guangdong province, South China, were investigated. Upwards the profile, δ44/40Ca values of the bulk saprolites systematically decrease from 0.77 ± 0.12 ‰ to -0.44 ± 0.12 ‰, suggesting that significant Ca isotope fractionation occurred during chemical weathering. The exchangeable fractions have δ44/40Ca values higher than those of the bulk saprolites with Δ44/40Caexchangeable-saprolite varying from -0.01 ‰ to 0.73 ‰, suggesting that heavy isotopes are preferentially adsorbed onto the clays. The acid-leachable phases display a relatively narrow δ44/40Ca range from 0.52 ‰ to 0.74 ‰ with Ca fractions varying from 7.4 % to 100.3 %, potentially indicating that limited Ca isotopic fractionation occurs during the dissolution of primary carbonates. The residual Ca pool is strongly fractionated with δ44/40Ca ranging from 0.64 ± 0.08 ‰ to -0.98 ± 0.02 ‰, systematically lower than their bulk saprolites, perhaps indicating light Ca isotopes are preferentially incorporated into the clay lattices. Altogether, it seems that the Ca isotopic fractionation directions are opposite between clay structural incorporation and adsorption. Our study provides important insight of Ca behavior and Ca isotopic fractionation during chemical weathering, which is critical to shape Ca isotopic compositions of the upper continental crust and trace the global biogeochemical cycle of Ca.

Source-specific ecological risk assessment and quantitative source apportionment of heavy metals in surface sediments of Pearl River Estuary, China.

In this study, surface sediments of the Pearl River Estuary were collected from 29 stations and investigated the spatial distribution, pollution level, quantitative source apportionment, and source-specific ecological risk of 10 heavy metals. The mean concentrations followed the order of Mn > Zn > Cr > Cu > Ni > Pb > As > Co > Cd > Hg. In terms of spatial distribution, it showed that the heavy metals were enriched in the inner Pearl River Estuary with 'extremely high' level of Hg, whereas, Cd and Zn posed 'moderate to high' contamination potential. We apportioned four main sources using positive matrix factorization model, in which natural geogenic and industrial manufacturing sources accounted for 36.84% and 27.11% of the total, respectively. However, the source-specific risk assessment suggested that mixed anthropogenic sources were the main contributors, and ecological risks were strongly affected by anthropogenic imports from the surrounding cities.

A multi-stable isotopic constraint on water column oxygen sinks in the Pearl River Estuary, South China.

Bottom water oxygen depletion is a central concern in estuaries and coastal oceans worldwide. However, a mechanistic understanding and quantitative diagnosis of different oxygen-consuming processes are less clear. In this study, a multi-stable isotope approach is developed to delineate the role of oxygen respiration and nitrification contributing to total oxygen consumption in the Pearl River Estuary (PRE), a large eutrophic estuary in south China. The approach highly couples with analysis of the carbon isotope composition of dissolved inorganic carbon (δ13C-DIC) and with stable nitrogen isotope analysis in ammonium (δ15N-NH4+) and nitrate (δ15N-NO3-). In all seasons, relatively low DO concentrations were observed in the upper reach and, to some extent, in the outer estuary during summer, while high concentrations of DO were found in the transition zone between the inner and outer estuary. On the basis of isotopic differentiation, our data reveal that much more depleted δ13C-DIC is coincident with DIC additions and low oxygen in the upper reach and inner estuary during most seasons. This is most likely a consequence of organic carbon (OC) degradation via aerobic respiration. Based on the carbon isotopic mass balance of DIC and the stoichiometry ratio of -ΔDO/ΔDIC, we found that the OC degradation dominates the total oxygen consumption in the upper reach, as well as in the inner estuary during summer (48.3%-93.5%). In addition, nitrification is another key process in contributing to total oxygen loss in the upper reach, as supported by the well-coupled variations of δ15N of NH4+ and NO3- and apparent oxygen utilization (AOU). Using the formerly determined N isotopic fractionation and observed δ15N variation, we estimated that nitrification could account for 35.3%-44.1% and 28.5%-31.6% of the total oxygen consumption in the upper reach during winter and summer, respectively, while its contribution to total oxygen loss is minor in the inner and outer estuary. Overall, this study demonstrates the potential of the multi-stable isotopic approach to assess oxygen sink partitioning in large human-perturbed estuaries.

Change of coral carbon isotopic response to anthropogenic Suess effect since around 2000s.

The stable carbon isotope composition (δ13C) in coral skeletons can be used to reconstruct the evolution of the dissolved inorganic carbon (DIC) in surface seawater, and its long-term declining trend during the past 200 years (~1800-2000) reflects the effect of anthropogenic Suess effect on carbonate chemistry in surface oceans. The global atmospheric CO2 concentration still has been increasing since 2000, and the Suess effect is intensifying. Considering the coral's ability of resilience and acclimatization to external environmental stressors, the response of coral δ13C to Suess effect may change and needs to be re-evaluated. In this study, ten long coral δ13C time series synthesized from different oceans were used to re-evaluate the response of coral carbonate chemistry to Suess effect under the changing environments. These δ13C time series showed a long-term declining trend since 1960s, but the declining rates slowed in eight time series since around 2000s. Considering that the declining rates of the DIC-δ13C in surface seawater from the Hawaii Ocean Time-series Station and Bermuda Atlantic Time-series Station has not changed since 2000 compared with those during 1960-1999, the change in the coral δ13C trends at eight of ten locations may indicate that the response of coral δ13C to the anthropogenic Suess effect has changed since around 2000s. This change may have resulted from coral acclimatization to external environmental stressors. To adapt to acidifying oceans, coral may have the ability to regulate the source of DIC in extracellular calcifying fluid and/or the utilization way of DIC, therefore the response of coral δ13C to anthropogenic Suess effect will change accordingly.

A Rapid and Simple Method for Lithium Purification and Isotopic Analysis of Geological Reference Materials by MC-ICP-MS.

A simple method has been developed to purify lithium (Li) from matrix elements in geological reference materials, using a single-column packed with AGMP-50 cation exchange resin, followed by high-precision Li isotope measurements by multi-collector inductively coupled plasma mass spectrometry (MC-ICP-MS). A series of tests, such as different types of resin, loading amount of Li, loading volumes, and various eluents, were conducted to ascertain the optimal conditions for Li purification and the effects of intensity, acidity, and presence of potential matrix elements on Li isotope measurements were also evaluated. In our experiment, Al and high-field-strength elements (HFSEs), such as Ti, Zr, and Hf, were eluted by 0.2 M HCl + 0.3 M HF, and 0.73 M HCl was used to separate Li from other matrix elements, such as Na. This method is suitable for processing large amount of Li (60-270 ng) and enabling a Li recovery of close to 100%, with effective removal of matrix elements such as Na and Ca. Besides, our method achieves low matrix interferences (e.g., Na/Li << 1 and Ca/Li << 1 for rock and seawater via a single-column procedure; Ca/Li < 2 for carbonate via a two-column procedure) and also uses small volume of eluents and is rapid (~5 h), enabling a total separation to be completed in ~0.5 d. Using this method, we report Li isotopic compositions of various geological reference materials, including igneous rocks, seawater, and carbonate. The Li isotopic compositions are consistent with the data published previously for the analyzed reference materials. As such, the reported method is ideally suited for Li separation from multiple types of geological samples prior to isotopic analysis.

Extreme weather events recorded by daily to hourly resolution biogeochemical proxies of marine giant clam shells.

Paleoclimate research has built a framework for Earth's climate changes over the past 65 million years or even longer. However, our knowledge of weather-timescale extreme events (WEEs, also named paleoweather), which usually occur over several days or hours, under different climate regimes is almost blank because current paleoclimatic records rarely provide information with temporal resolution shorter than monthly scale. Here we show that giant clam shells (Tridacna spp.) from the tropical western Pacific have clear daily growth bands, and several 2-y-long (from January 29, 2012 to December 9, 2013) daily to hourly resolution biological and geochemical records, including daily growth rate, hourly elements/Ca ratios, and fluorescence intensity, were obtained. We found that the pulsed changes of these ultra-high-resolution proxy records clearly matched with the typical instrumental WEEs, for example, tropical cyclones during the summer-autumn and cold surges during the winter. When a tropical cyclone passes through or approaches the sampling site, the growth rate of Tridacna shell decreases abruptly due to the bad weather. Meanwhile, enhanced vertical mixing brings nutrient-enriched subsurface water to the surface, resulting in a high Fe/Ca ratio and strong fluorescence intensity (induced by phytoplankton bloom) in the shell. Our results demonstrate that Tridacna shell has the potential to be used as an ultra-high-resolution archive for paleoweather reconstructions. The fossil shells living in different geological times can be built as a Geological Weather Station network to lengthen the modern instrumental data and investigate the WEEs under various climate conditions.

Carbon and oxygen isotopic analyses of calcite in calcite-dolomite mixtures: Optimization of selective acid extraction.

RATIONALE: Carbon and oxygen isotopic compositions of individual minerals in calcite-dolomite mixtures could be measured using a selective acid extraction technique based on the different reaction rates of calcite and dolomite with phosphoric acid. However, poor accuracies of calcite are usually obtained when mixtures of low calcite content are analyzed, which may result in incorrect conclusions. To overcome this shortcoming, improvements are needed in accuracy and precision when using the technique with mixtures of low calcite content. METHODS: Calcite and dolomite were analyzed under different reaction conditions using a GV Isoprime™ II isotope ratio mass spectrometer equipped with a dual inlet (DI) and an automatic sampler, to study the relationships between isotopic compositions and CO2 yield, and reaction time and temperature. Artificial mixtures with varied calcite:dolomite ratios were prepared, and their calcite isotopic compositions were determined with the aim of optimizing analytical conditions. RESULTS: The CO2 yields and isotopic compositions of calcite and dolomite were obtained on-line under different experimental conditions using DI-isotope ratio mass spectrometry (DI-IRMS). Based on the analysis results of artificial mixtures, the reaction conditions for analyzing isotopic composition of calcite in mixtures were optimized as follows. Samples should have grain sizes of 75-80 µm; and samples with calcite contents of >50%, 30-50% and < 30% should be reacted with acid for 10-15 min at 50°C, 6 min at 50°C and 45 min at 25°C, respectively. CONCLUSIONS: Optimized analytical conditions are suggested for the determination of the isotopic compositions of calcite in mixtures with improved accuracy by an automatic selective acid extraction technique.

Differentiation analysis of boron isotopic fractionation in different forms within plant organ samples.

As a critical micronutrient, boron (B) plays an important role in plant growth and embryonic development. To further understand the effects of B uptake, transportation and isotopic fractionation, the contents and isotopic compositions of hydro-soluble B in the sap and structural B fixed in the cell within individual plant tissues were investigated. The B isotope ratio was determined by multi-collector inductively coupled plasma mass spectrometry. The δ11B values in hydro-soluble and structural B in the investigated plant samples ranged from -1.57‰ to +11.30‰ and from +6.57‰ to +16.64‰, respectively. Different fractionation factors of the B isotopes, in the range of 0.9954-1.0150, were observed in these samples, indicating that in most plant tissues, the heavy isotope (11B) was preferentially enriched in structural B, which was fixed into the cell. However, there was a reversal in the fractionation of B isotopic compositions in the fruit samples compared with the other plant tissue samples. It is more powerful to examine the molecular mechanisms of B transport, uptake and utilization than the use of limited plant organ samples containing a mixture of hydro-soluble and structural B within different intra-plant compartments and in inter-plant interactions. These isotopic shifts, which may be used as important isotopic indicators, contribute to the surface processes interactions in the plant-soil system and the knowledge of the molecular mechanisms of B in the uptake and absorption by different plant species in nature.

Purification of BaSO4 precipitate contaminated with organic matter for oxygen isotope measurements (δ18 O and Δ17 O).

RATIONALE: Sulfate precipitates are often contaminated with nitrates and organic materials (OM), which reduce the precision and accuracy of measurements of δ18 O and Δ17 O values in the sulfate. Although nitrates can be effectively removed using diethylenetriaminepentaacetic acid solution, removing OM from the precipitates is often difficult. One effective approach is to heat powdered precipitates to high temperatures. In this study, the effect of this procedure on the δ18 O and Δ17 O values of BaSO4 precipitate was fully examined. METHODS: OM-contaminated BaSO4 precipitates and 18 O- and 17 O-labeled purified BaSO4 precipitates were loaded into alumina and gold crucibles and heated at 450 °C, 600 °C and 800 °C for 2 h. The nitrogen and carbon contents in the initial and the final BaSO4 were measured using an elemental analyzer. The values of δ18 O and Δ17 O were measured using a temperature conversion/elemental analyzer coupled with an isotope ratio mass spectrometer, and a CO2 laser system coupled with an isotopic ratio mass spectrometer. RESULTS: OM was effectively (88 ± 17%) removed from the BaSO4 precipitates by this treatment, and heating at 800 °C had the highest removal efficiency (98%). The differences in δ18 O and Δ17 O values between the final and initial BaSO4 precipitates was -0.6‰ to 0.3‰ (average of -0.1‰) and -0.24‰ to 0.10‰ (average of -0.02‰), respectively. Significant positive relationships between the δ18 O and Δ17 O values of the initial BaSO4 precipitate and those of the high-T-treated aliquots were found, with slopes having mean values of 0.96 ± 0.06 and 1.04 ± 0.01, respectively. CONCLUSIONS: The result demonstrates high removal efficiency for OM in BaSO4 precipitates and no significant differences in the oxygen isotopic compositions between high-T treated BaSO4 and initial BaSO4 . This study indicates that the modified high-T treatment (800 °C, 2 h) is an effective method for purifying BaSO4 precipitated from geological and environmental samples with a high OM content for δ18 O and Δ17 O measurements. Copyright © 2016 John Wiley & Sons, Ltd.

Acceleration of modern acidification in the South China Sea driven by anthropogenic CO2.

Modern acidification by the uptake of anthropogenic CO2 can profoundly affect the physiology of marine organisms and the structure of ocean ecosystems. Centennial-scale global and regional influences of anthropogenic CO2 remain largely unknown due to limited instrumental pH records. Here we present coral boron isotope-inferred pH records for two periods from the South China Sea: AD 1048-1079 and AD 1838-2001. There are no significant pH differences between the first period at the Medieval Warm Period and AD 1830-1870. However, we find anomalous and unprecedented acidification during the 20th century, pacing the observed increase in atmospheric CO2. Moreover, pH value also varies in phase with inter-decadal changes in Asian Winter Monsoon intensity. As the level of atmospheric CO2 keeps rising, the coupling global warming via weakening the winter monsoon intensity could exacerbate acidification of the South China Sea and threaten this expansive shallow water marine ecosystem.

Genotoxicity of the sediments collected from Pearl River in China and their polycyclic aromatic hydrocarbons (PAHs) and heavy metals.

The accelerated industrialization and urbanization in the last three decades around the Pearl River Delta within Guangdong Province in China have led to serious concerns about the impacts on the aquatic environment. In the present study, the genotoxicity of the sediments collected from the Pearl River was evaluated by micronucleus (MN) assay with Vicia faba root tip cells, and the 16 EPA priority polycyclic aromatic hydrocarbons (PAHs) and heavy metals (HMs, including Cr, Cu, As, Se, Cd, Hg, and Pb) in the sediments were determined respectively by GC-MS, inductively coupled plasma mass spectrometry, and inductively coupled plasma atomic emission spectrometry. The results showed that there were significant increases of MN frequencies observed in the sediment-exposed groups, compared with the negative group (P < 0.05, P < 0.01), indicating that the sediments clearly had genotoxicity to the V. faba root cells. The total concentrations of the priority PAHs (250-13,656 ng g(-1), dry weight) and HMs (As, 22,770-36,639 µg kg(-1); Cr, 39,333-133,343 µg kg(-1); Cu, 36,145-159,270 µg kg(-1); Pb, 51,210-166,642 µg kg(-1); Cd, 475.4-1,818.9 µg kg(-1); Hg, 59.9-460.8 µg kg(-1); and Se, 331.7-1,250.4 µg kg(-1), dry weight) were close to those obtained from other urbanized and industrialized areas, which have been considered moderately polluted. There was a clear positive correlation between MN potency and the molar concentrations of Hg and Pb in the sediments (Hg, r = 0.94; Pb, r = 0.91), suggesting that Hg and Pb were the most important factors that posed the sediments higher genotoxicity to V. faba root cells. Our results suggested that both biological and chemical approaches are necessary to be included in a battery of tests to assess the eco-environmental risks of sediments.

Interannual variation of rare earth element abundances in corals from northern coast of the South China Sea and its relation with sea-level change and human activities.

Here we present interannual rare earth element (REE) records spanning the last two decades of the 20th century in two living Porites corals, collected from Longwan Bay, close to the estuarine zones off Wanquan River of Hainan Island and Hong Kong off the Pearl River Delta of Guangdong Province in the northern South China Sea. The results show that both coral REE contents (0.5-40 ng g⁻¹ in Longwan Bay and 2-250 ng g⁻¹ in Hong Kong for La-Lu) are characterized with a declining trend, which are significantly negative correlated with regional sea-level rise (9.4 mm a⁻¹) from 1981 to 1996 in Longwan Bay, 13.7 mm a⁻¹ from 1991 to 2001 in Hong Kong). The REE features are proposed to be resulted from seawater intrusion into the estuaries in response to contemporary sea-level rise. However, the tendency for the coral Er/Nd time series at Hong Kong site is absent and there is no significant relation between Er/Nd and total REEs as found for the coral at Longwan Bay site. The observations are likely attributed to changes of the water discharge and sediment load of Pearl River, which have been significantly affected by intense human activities, such as the construction of dams/reservoirs and riverbed sediment mining, in past decades. The riverine sediment load/discharge ratio of the Pearl River decreased sharply with a rate of 0.02 kg m⁻³ a⁻¹, which could make significant contribution to the declining trend of coral REE. We propose that coastal corals in Longwan Bay and similar unexplored sites with little influences of river discharge and anthropogenic disruption are ideal candidates to investigate the influence of sea-level change on seawater/coral REE.

Heavy metal pollution recorded in Porites corals from Daya Bay, northern South China Sea.

We examined metal-to-calcium ratios (Fe/Ca, Mn/Ca and Zn/Ca) in the growth bands of two Porites corals from Daya Bay, South China Sea, in order to trace long-term trends in local ambient pollution levels. Although Fe and Mn did not show any obvious increasing trends over 32 years in the period 1976-2007, peak values of Fe/Ca and Mn/Ca occurred in the mid-late 1980s, temporally-coeval with the local construction of a nuclear power station. Furthermore, both corals showed rapid increases in Zn concentrations over the past 14 years (1994-2007), most likely due to increases in domestic and industrial sewage discharge. The Daya Bay corals had higher concentrations of metals than other reported corals from both pristine and seriously polluted locations, suggesting that acute (Fe and Mn) and chronic (Zn) heavy metal contamination has occurred locally over the past approximately 32 years.

Leachates of municipal solid waste incineration bottom ash from Macao: heavy metal concentrations and genotoxicity.

Heavy metals in municipal solid waste incineration bottom ash (MSWIBA) may leach into soil and groundwater and pose long-term risks to the environment. In this study, toxicity characteristic leaching procedure (TCLP) was carried out on the MSWIBA from Macao. Heavy metals in leachates were determined by inductively coupled plasma mass spectrometry (ICP-MS) and inductively coupled plasma atomic emission spectrometry (ICP-AES), and genotoxicity of leachates was also evaluated by micronucleus (MN) assay with Vicia faba root tip cells. The results showed that the concentrations of aluminium (Al), manganese (Mn), cobalt (Co), cadmium (Cd) and mercury (Hg) in the leachates were less than 0.01 mg l(-1), and those of iron (Fe), copper (Cu) and molybdenum (Mo) were less than 0.1 mg l(-1). The concentrations of chromium (Cr), zinc (Zn), selemium (Se), strontium (Sr), barium (Ba) and caesium (Cs) were between 0.11 mg l(-1) and 2.19 mg l(-1). Lead (Pb) concentrations, in particular, reached as high as 19.6 mg l(-1), significantly exceeding the maximum concentration limit (5 mg l(-1) for lead by TCLP). Compared with the negative group, a significant increase of MN frequencies was observed in the leachate-exposed groups (P<0.05). With the increase of heavy metals in the leachates, the toxic effects on the Vicia faba root tip cells increased, implying that heavy metals were the main factors causing the genotoxic effects. Our results suggested that apart from chemical analysis, bioassays like the MN assay of Vicia faba root tip cells should also be included in a battery of tests to assess the eco-environmental risks of bottom ashes before decisions can be made on the utilization, treatment or disposal.