Ecology and Environmental Protection

ECOLOGICAL ASSESSMENT OF MOBILE CADMIUM IN SEDIMENTS

FROM CERTEJ MINING SITE, HUNEDOARA COUNTY, ROMANIA

Dr. Gabriela Geanina Vasile

Dr. Lidia Kim
Dr. Stefania Gheorghe
Bogdan Stanescu
National Research and Development Institute for Industrial Ecology, Romania

ABSTRACT
The aim of this paper is to assess the impact on the aquatic environment due to
cadmium pollution from Certej mining site from Romania.
The total and mobile concentrations of Cd were analyzed in 30 sediment samples and
also 30 surface water samples collected from Certej catchment area (Hunedoara County,
Romania) in three sampling campaignes performed in 2012.
The total and mobile Cd2+ were analyzed from sediment samples using flame atomic
absorption spectrometry. Selection of the most appropiate method for estimation of
mobile Cd was performed using a matrix type certified reference material (BCR 701)
and were tested for different extraction methods. Total cadmium values were situated in
the range between 1 and 7.3 mg/kg dry matter, exceeding the maximum admissible
value, which is 0.8 mg/kg in Romania Legislation. The acid pH of water and sediment
samples increase the mobility of metal species that will be found in dissolved forms in
surface water. Leaching fraction percentages indicates a high mobility of Cd2+ in
sediment samples, mostly situated above 70%.
In order to estimate the impact of the highest concentration of Cd 2+ occured in Certej
sediments on the living organisms, ecotoxicological studies were performed. The Cd 2+
toxicity was measured using a bioassay batery with fish (Cyprinus carpio), planktonic
crustaceean (Daphnia magna) and luminiscent bacteria (Vibrio fischery). The results
showed a major toxicity on all tested organisms, with L(E)C 50<1 mg/l which indicated
an adverse impact on aquatic biota.

Keywords: sediment, cadmium, mining site, mobility, ecotoxicity

INTRODUCTION
Sediments represents an important compartment in aquatic ecosytems and also one of
the ultimate absorbent for heavy metals discharged into the aquatic environment. It
constitues a habitat for many organisms and is an essential throphic resourse for higher
level organisms.
Persistent and pottentially harmful products have been widely disperded and
concentrated in water, sediment and living organism as result of human activities
(nutrients load from agricultutal fertilizers and heavy metals from industry such as
mining). Contaminants are not necessarily fixed permanently by the sediments, and

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under changing environmental conditions metals may be released to the water column
by various processes of remobilization. Thus, in aquatic ecosystems, sediments may be
both a carrier and possible source of pollutants [1].
The total concentration of metals does not provide suitable information on the
bioavailability of hazardous elements and the speciation study is a necessary tool to
obtain this information [2]. Many leaching tests, often-single procedures, have been
developed and implemented to evaluate the reactivity of metals in sediments. For
sediments and suspended particles, the use of selective leaching methods can provide
key informations on the nature of metal species bound to the solid phase such as:
exchangeable ions; carbonates, with some iron and manganese oxides; readily reducible
species (certain oxides); oxidisable organics and sulphide [3]. As the final step, a total
digestion with a mixture of strong acids could provide the lithogenous fraction [4, 5].
Cd2+ toxicity was studied in many papers due to their extensive usage, globally about 19
700 tones were obtained from mining in 2000 and 62% from total industrial production
are evacuated through wastewater contaminating the surface waters. In European
continental waters Cd2+ is founded in the range of 10-100 mg/L [6].
This metal, alone or in combination with other metals or organic compounds, develop
adverse effects on aquatic organisms emphasized though vital functions disturbance and
bioaccumulation [7, 8].
In this context, the aim of this paper is to assess the impact on the aquatic environment
due to cadmium pollution from Certej mining site Hunedoara county, Romania.

MATERIALS AND METHODS

Collection, analysis and processing of water and sediment samples
Thirty sediment samples and also surface water samples in contact with sediments were
collected from ten points situated in Certej catchment area (Hunedoara County,
Romania) in three sampling campaignes performed in 2012 (February till April). The
schematic map of mining site is presented in figure 1 and the description of sampling
points are in table 1.
Water samples were collected and stored in polyethylene bottles. From each sampling
location, 10 L of water were collected. The sediment samples were taken using a Van
Veen Bottom Sampler and collected in glass bottles. All samples were kept in cooling
boxes at 40 C during transportation and the analyses were performed immediately after
receiving the samples in the analytical laboratory of Evaluation, Monitoring and
Environmental Pollution Department from National Research and Development
Institute for Industrial Ecology, Bucharest, Romania.
A part of sediment samples was dried on laboratory temperature, homogenized, crushed
to fine particles and sieved in a Fritsch Analysette 3 Spartan Vibratory Sieve Shaker and
the fraction less than 63 µm was collected for evaluation of the total content of
cadmium. The other part of sediment samples, in order to maintain the same structure as
in natural conditions, was sieved in wet condition using surface water from same
sampling point and particle size less than 63 µm was collected.

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Sediment extractions - total content and mobile fraction

A subsample of each dry sediment sample (particle size less then 63 µm) was dissolved
with a mixture of strong mineral acids ( HCl:HNO 3 = 1:3) in a Microwave Laboratory
Ethos System, so that the total concentrations of Cd were obtained.

Figure 1. Map of sediments sampling points – Certej aria

Table 1. Description of sampling points from Certej mining site

Code Description of the sampling points
P1 Sampling point on Coranda Spring - upstream
P2 Sampling point situated on Baiaga Spring, before crossing with Coranda Spring
P3 Sampling point situated at the exit of Nicodim gallery (infiltration water, red-orange color)
Sampling point situated on Baiaga Spring, downstream of the confluence with water from
P4
gallery Nicodim
P6 Sampling point situated on Baiaga Spring, upstream of the confluence with career water
P6 Sampling point from career water, before confluence with Baiaga Spring (reduce flow)
P8 Sampling point from Ciongani Spring before crossing with Baiaga Spring
Sampling point on Baiaga Spring situated upstream of the confluence with Ciongani Spring
P7
and downstream of the confluence with career water
P9 Sampling point on Baiaga Spring, downstream of the confluence with Ciongani Spring
P10 Sampling point from Baiaga Spring in Hondol village.

Selection of the most appropiate method for estimation of mobile Cd 2+ was performed
using a matrix type certified reference material, lake sediment BCR 701.
In experimental study were tested four different extraction methods (table 2) in order to
evaluate the mobile fraction of Cd [9, 10]. Methods 3 and 4 represents first step of BCR
extraction procedure, nothing that method 4 uses a ultrasonic bath instead of room
temperature and so, decreases the time required for the extraction from 16 hours to 20
minutes [11].
The single chemical extraction procedure (figure 2) was performed three times on each
sediment sample, the final results representing the average. Also, blank extraction (i.e.,
without sediment) was carried out for each set of analysis, using the same reagents.

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Figure 2. Flow scheme for single chemical extraction of mobile Cd

Table 2. Single chemical extraction procedures applied on dry sediments
Methods Method 1 Method 2 Method 3 Method 4
Reagents 0.01 M EDTA, TEA 0.1 mol/L; CH3COOH CH3COOH
1 M CH3COONH4CaCl2 0.01 mol/L; 0.11 mol/L 0.11 mol/L
pH=7±0.2 DTPA, 0.005 mol/L;
pH=7.3
Condition 20ºC, 20ºC, 30 ± 3 rot/min, 20ºC, 30 ± 320ºC, 20 minutes,
40±4rot/min, report report rot/min, ultrasonic bath, 35 kHz.,
sediment: extractionsediment: extractionreport report
solution =1:10 (m/v),solution = 1:10 (m/v), 2sediment: sediment: extraction
2 hours hours extraction solutionsolution= 1:40 (m/v).
= 1:40 (m/v), 16
hours

The sediment dry weight was determined on a separate subsample and a correction to
„dry matter” (quantity of metal per g dry sediment) was applied to all analytical results
(total content of Cd). In case of wet sediments, a dry matter corrections was performed
before the suspensions were prepared, in order to maintain same ratio between
sediment and solution.
Analytical technique used for determination of cadmium from surface water and
sediment samples (total and mobile fraction) was flame atomic absorption spectrometry
performed on THERMO SCIENTIFIC M6 DUAL SOLAAR Series, using air/acetylene
type flame. All the chemicals were of analytical reagent grade (Merck quality). Matrix
of the solution used for calibration curve matched with the extraction solutions (table 2)

Toxicity tests
Aquatic impact assessment of cadmium (the principal inorganic pollutant from the
Certej sediments) was revealed. Three acute toxicity bio tests were performed
considering the OECD and ISO methodology and using the micro bio tests [12], in
order to evaluate the effects on freshwater fish, planktonic crustaceans and luminescent
bacteria. In table 3 are summarized the tests conditions and the range of tested
concentrations.
The experiments were conducted for a synthetic Cd 2+ solution (using CdSO4) which
simulate the highest concentration (7 mg/L) recorded in collected sediment samples. In

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figures 3, 4 and 5 are presented the testing equipment’s used in toxicity assessment from
Aquatic Biobase of National Research and Development Institute for Industrial
Ecology.
Estimation of toxicity values was performed using the probity analysis method (in case
of fish and bacteria) and exponential regression model (for bacteria). All the tests were
carred out in the same time with a control test and a reference test with potasium
dicromate.

Figure 3. Fish toxicity Figure 4. Crustaceans Figure 5. Bacteria

testing aquaria toxicity experiment design toxicity equipment

Table 3. Tests conditions

Type of Test Range of tested
Bioassay Species Endpoints
test period concentrations
Serial dilutions
Cyprinus 96h, Mortality and
OECD 203 acute 0.015 – 1.7 mg/L;
carpio 21-22oC behavior, LC50
7 mg/L
OECD 202 Serial dilutions
Daphnia 24-48h, Mortality and
Daphtoxkit F acute 0.015 -1 mg/l; 7
magna 20oC behavior, EC50
magna mg/l
DIN EN ISO
Luminescence
11348-3 Vibrio 15 min, Serial dilutions
acute inhibition,
BioFix Lumi, fischeri 20oC 0.2 - 7 mg/L
IC50
Multi-Shot

RESULTS AND DISSCUTIONS

Coranda and Ciongani Springs represents receptors for percolated water from career and
sterile minig waste dump. Carrer water colects mine water and washing water dumps
and represent an important sourse of contamination of the springs.
Pollution in surface water is strongly dependent by the water flow that is in contact with
rocks / sediments and causes leaching of the metals. Depending on the amount of
rainfall in the area, surface water can be loaded or diluted.
First sampling activity was performed after difficult winter, when a large amount of
snow fall, ice was formed and surface water flow was very low.
However, it can be hightlight some general characteristics of the area, such as: acid pH
values, high content of total metals (As, Cd) and mobile metals (Cd, Fe, Mn, Zn).
Comparing the results with the limits imposed by the Romanian Order no. 161/2006
[13] for surface water quality, the pH values were within the acid range, around 3.5 pH
unit, outside the set points. The quality of surface water was situated in 5 th class
represented the wors quality, in entire study area in all months.
Cadmium values recorded both in sediment and surface water have a momentarily
character, associated with location and time of sampling (dry season or rainfalls).

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Total cadmium values in sediment samples were situated in the range between 1 and 7.3
mg/kg dry matter, exceeding the maximum admissible value, which is 0.8 mg/kg in
Romania Legislation. The limit value of cadmium was exceeded from 1.6 to 8.3 times
in March and was situated betwwen 2.7 to 8.8 times higher in April.
Experimental results indicated that classical method BCR (method 3, table 2) and
modified method BCR (method 4) applied to CRM BCR 701 conducts to similar
results. Thus, considering the operating time, method 4 was used in order to extract
mobile fraction od Cd from Certej area sediments.
The acid pH of water and sediment samples increase the mobility of metal species that
will be found in dissolved forms in surface water (figure 3).
Leaching fraction percentages indicates a high mobility of Cd 2+ in sediment samples,
mostly situated above 70% (figures 4, 5 and 6).

Figure 3. Variation of pH in sediment Figure 4. Variation of total and mobile Cd

samples in first campaign

Figure 5. Variation of total and mobile Cd Figure 6. Variation of total and mobile Cd
in second campaign in third campaign
Toxicity assessment

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The laboratory toxicity effects were presented in table 4. Results showed an acute lethal
toxicity of Cd2+ on all tested aquatic organisms. A highly toxic effect on Daphnia
magna and Vibrio fischeri bacteria was revealed. According to other Cd2+ toxicity data,
the estimated values of CL50/EC50 are in the literature ranges.
Many microbial studies on metals toxicity highlighted hazardous effects on the marine
bacteria and low toxicity on the freshwater and sludge bacteria as a result of organic
pollutants and salts loads in the aqueous matrix which reduce the metals bioavailability
at bacterial membranes modifying their toxicity level [6]. Considering that tested
synthetic solution of cadmium had no other organic constituents, the showed toxicity (1
mg / L <LC50<10 mg / L) is the direct effect of metal which increased their
bioavailability at bacterial membranes.
Acute toxicity values determined for fish (juvenile carp, Cyprinus carpio) and
phytoplanktonic crustaceans (Daphnia magna) indicate a strong toxicity (LC50 / EC50 <1
mg / L), which reveled that the Cd2+ have the ability to cause harm on the aquatic
environment in a short and long time.
Table 4. Cadmium aquatic toxicity – EC50/LC50, NOEC, LOEC, literature data and
toxicity class
Species EC50/LC50 (mg/L) NOEC (mg/L) LOEC (mg/L)
Daphnia magna 0.14 0.015 0.03
Vibrio fischeri 3.4 0.1 0.2
Cyprinus carpio 0.16 0.01 0.02
Highly Toxic for fish and crustaceans
Toxicity class according to EPA
Toxic for bacteria
Fish (Carassius auratus, Rainboow trout, Brown trout,
Lepomis macrochirus, Pimephales promelas, Poecilia
Literature ranges
reticulata, Brachydario rerio) LC50 = 0.015 - 4.56 mg/L;
(IUCLID 2000,
Crustacens (Daphnia magna, Daphnia pulex, Daphnia
EaSI – View)
hyalina) EC50 = 0.04 – 0.1 mg/L;
Bacteria (Salmonella typhimurium) IC50 = 10.4 mg/L;
Note: LC50/EC50 – Lethal / Effective Concentration for 50% of tested organisms (crustacean, fish,
bacteria); NOEC – No Effect Concentration; LOEC – LOwest Effect Concentration.

CONCLUSIONS
Total cadmium values in sediment samples collected from Certej mining site were
situated in the range between 1 and 7.3 mg/kg dry matter, exceeding the maximum
admissible value, which is 0.8 mg/kg in Romania Legislation.
The acid pH of water and sediment samples increase the mobility of metal species that
will be found in dissolved forms in surface water. Leaching fraction percentages
indicates a high mobility of Cd2+ in sediment samples, mostly situated above 70%.
In order to estimate the impact of the highest concentration of Cd 2+ occured in Certej
sediments on the living organisms, ecotoxicological studies were performed. The Cd 2+
toxicity was measured using a bioassay batery with fish (Cyprinus carpio), planktonic
crustaceean (Daphnia magna) and luminiscent bacteria (Vibrio fischery). The results
showed a major toxicity on all tested organisms, with L(E)C 50<1 mg/l which indicated
an adverse impact on aquatic biota.

REFERENCES

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