Romanian Journal of Ecology & Environmental Chemistry
Romanian Journal of Ecology & Environmental Chemistry
Romanian Journal of Ecology & Environmental Chemistry
NO. 1
Romanian Journal of
Ecology & Environmental Chemistry
BUCHAREST, ROMANIA
SCIENTIFIC COMMITTEE
www.incdecoind.ro
Edited by EIKON
INTRODUCTORY WORDS
“The concerns regarding the sustainability of our influence on the environment are steadily
increasing even there is great progress in scientific understanding of human interference with nature. Also
technology development has made great progress in solving environmental problems. Environmental
Chemistry is a key discipline in both areas in science as well as in technology. In order to meet the
challenges coming from the UN Sustainable Development Goals we urgently need an improved insight
into the close relationship between human development and the natural environment which is becoming a
primary task of Ecology in the future. In fact, ecology is a very complex scientific area where many
scientists of different disciplines must work together in order to improve our understanding on how we will
have to change our behaviour in order to achieve the SDGs. Industry is playing a dominant role in two
ways: industry is one of the main users of natural resources and key player in materials transformation to
products and waste material which are released to the environment water, air and soil. At the same time
industry is still a major source of jobs and is markedly contributing to increasing our wellbeing, health
and life expectance.
The 17 goals are all interrelated and wherever quality criteria play an important role
Environmental Chemistry is one of the primary disciplines to quantify the actual flow of materials
through our society and allowing to measure the progress in the future. The new Journal has the goal to
enhance the international scientific knowledge transfer as well as to promote innovations contributing to
achieve the SDGs with a focus on industrial development based on sound science and proven technological
experience which both are also an important basis for political decisions.”
Em. O. prof. dr. Helmut KROISS
Vienna University of Technology
“I am pleased to hear about the creation of „Romanian Journal of Ecology & Environmental
Chemistry”. Unfortunately, the present development of our civilization is inevitably associated with the
production of goods (often trashy or useless). A side effect of such development is the devastation of the
natural environment including the chemical pollution, especially dangerous for living organisms. Thus the
setup of the new journal devoted to chemical issues of environmental protection should be considered a
highly desirable undertaking. I wish the Editorial Team of the „Romanian Journal of Ecology &
Environmental Chemistry” many significant articles helpful in solving ecological problems”.
Prof. dr. Stanislaw KOTER
Nicolaus Copernicus University in Toruń, Poland
“Environmental issues, including global warming, water, air and soil pollution, are major topics
for discussion both globally and locally. Throughout time there has been a stable balance in nature. There
is a harmony between biotic and abiotic factors in the environment. Since the 1950s as a result of
advances in industry, agriculture, and everyday life, increasing amounts of waste have accumulated in the
environment. Gases, liquids and solid waste, including domestic and waste water, pollute environment.
Where biodegradable chemical substances, from the use of polluting technologies for the production
of material goods, irrational use of pesticides and mineral fertilizers in the agrarian sector, household
waste etc. penetrate into the environment, natural balance is lost and this imbalance can have disastrous
effects on life on Earth.
Results of studies of processes and mechanisms of transformation, migration, chemical and
biochemical decomposition, immobilization, oxido-reduction, adsorption, etc. of organic and inorganic
pollutants entering environmental objects will be published in the new Journal of Ecology and
Environmental Chemistry founded by INCO-ECOIND and will form the basis for monitoring,
assessment and control of pollution of the environment, development of efficient chemical, biochemical and
biological technologies for environmental treatment. The journal will have a positive impact on the
ecological education of the population, and especially on the younger generation.”
Acad., Prof. Dr. Habilitat, Tudor LUPASCU
Institute for Chemistry, Academy of Science Moldova
CONTENT
pag.
National R&D Institute for Industrial Ecology – ECOIND: “42 years of excellence in
environmental research and services”………………………………………………................... 1
Elena Bucur, Valeriu Danciulescu, Gheorghita Tanase, Carol Blaziu Lehr, Andrei
Vasile, Odour pollution assessment through indirect methods based on the monitoring of
technological parameters – case study………………………………………………………….. 4
Mirela Alina Constantin, Lucian Alexandru Constantin, Ines Nitoi, Ionut Cristea, Ellea
Boulac, Photo-degradation of 5-fluorouracil. Comparative study on the efficiency of
UV/H2O2, UV/TiO2, UV/H2O2/TiO2 processes………………………………………………… 11
Stefania Gheorghe, Irina Lucaciu, Iuliana Paun, Gabriela Geanina Vasile, Jana Petre,
Vasile Ion Iancu, Catalina Stoica, Daniel Mitru, Mihai Nita-Lazar, Anthropogenic
pollutants: 10 years of progress in ecotoxicological studies and aquatic risk
assessment………………………………………………………………………………………. 26
Ines Nitoi, Lucian Alexandru Constantin, Ionut Cristea, Mirela Alina Constantin,
Advanced oxidation processes (AOPs) alternative methods for degradation of toxic pollutants
from wastewater………………………………………………………………………………… 39
Jana Petre, Vasile Ion Iancu, Toma Galaon, Marius Simion, Marcela Niculescu,
Gabriela Geanina Vasile, Luoana Florentina Pascu, Carol Blaziu Lehr, Liliana Cruceru,
Simona Calinescu, Gabriel-Lucian Radu, Occurrence of pharmaceutical and disinfectants
in the dissolved water phase of the Danube River and three major tributaries from
Romania………………………………………………………………………………………… 55
Bogdan Stanescu, Adriana Cuciureanu, Soil management in the urban environment: today
and future……………………………………………………………………………………….. 66
Catalina Stoica, Elena Stanescu, Iuliana Paun, Alina Banciu, Stefania Gheorghe, Irina
Lucaciu, Gabriela Geanina Vasile, Mihai Nita-Lazar, Danube Delta: monitoring and
ecological status. A link between the past and the future ……………………………………… 72
National R&D Institute for Industrial Ecology – ECOIND: “42 years of excellence in
environmental research and services”
The institute was founded in 1977 under the the institute continuously evolved from a
name of ICPEAR (Institute of Research and research institute dedicated to development of
Design for Wastewater Treatment) with technologies and design of installations for
headquarter located in Bucharest, one subsidiary treatment of wastewater generated by chemical
in Timisoara and later in 1982 another and petrochemical industries to a national
subsidiary was established at Ramnicu Valcea, research and development institute, with o
having as activity domain: technological global approach of environmental aspects in the
development and design of installations for field of industrial ecology, well known at
treatment of wastewater generated by chemical national level and with a growing presence
and petrochemical industries. within international partnerships.
Due to its unicity, obtained results, ECOIND was reaccredited as national institute
multidisciplinary approaches and adequate in 2001, 2008 and 2012 by the Ministry of
response to the environmental issues of Education and Research and by National
economic units, in 1999 ICPEAR was Agency for Scientific Research respectively.
transformed through accreditation into National Nowadays, ECOIND is operation under
Research and Development Institute for coordination of Ministry of Research and
Industrial Ecology – NR&DI ECOIND (GD no. Innovation (GD 13/2017) and represents the
261/1999 and Regulation of Organization and only institute at national level with a global
Operation approved by GD no. 961/2005). approach of environmental issues in the field
From its establishment (1977) until present days, of industrial ecology.
We consider that the new scientific journal „ROMANIAN JOURNAL OF ECOLOGY &
ENVIRONMENTAL CHEMISTRY”, next to International Symposium “The Environment and
The Industry” now at the 22 edition and institutional repository “Library of National Research and
Development Institute for Industrial Ecology ECOLIB” developed since 2017, will contribute to
promotion of the obtained results both at national and international level, in the field of industrial
ecology and environmental chemistry.
No matter how many barriers we’ll have to overcome our motto will remain the same:
“EXCELLENCE IN ENVIRONMENTAL RESEARCH AND SERVICES”
ECOIND Team
The odours in the ambient air, through the discomfort that they induce, represent an actual problem
for the communities located nearby companies with technological processes that emit in the air
different strong and unpleasant odorous substances.
The standard method for odour assessment involves measuring the concentration through dynamic
olfactometry according with SR EN 13725:2003, a very complex method that requires specialized
working staff and expensive equipment.
The paper presents an indirect method for odour assessment in the ambient air, based on
monitoring the process and meteorological data - Predictive Emission Monitoring Systems (PEMS)
and it is applied to a livestock farm. Using the multiple regression analysis of the monitoring data
for the most important specific technological and meteorological parameters it can be developed a
mathematical model that could be used for the calculation of odour concentration in air, without
the necessity of direct measurement, after the initial step. For the case study presented in the paper,
the distance between the slurry lagoon was identified as a significant statistical parameter that can
determine in a proportion of 72% the concentration of odour in the ambient air nearby the farm;
the margin of error for odour concentration assessment, according to the model validation tests, is
± 8%, acceptable value for an estimation method by mathematical modelling.
Keywords: odour, dynamic olfactometry, multiple regression method, intensive livestock farming
Odour can be defined as sensations resulting indirect methods can be used, methods based on
from the interaction of volatile chemical species statistical analysis of the parameters values that
inhaled through nose, making contact with the are correlated with the level of odour emission.
olfactory area and registering in the brain [1]; in In this regard, the use of statistical correlation
this context, the standardized method for odour and regression methods have been applied in
determination, the method based on dynamic various activity, including environmental
olfactometry [2-4] uses the human nose, protection. [6-10].
respectively a group of odour assessor connected The paper presents an indirect method for odour
to an olfactometer – a performant system that assessment based on the monitoring of process,
dilutes, present the samples to the human meteorological and geographical parameters -
assessors, and does all the calculations using a Predictive Emission Monitoring Systems [11] –
data processing software. An automatic applied to a livestock farm. Using the multiple
alternative based on electrochemical cells is regression analysis applied to specific
represented by the electronical nose, used monitoring parameters data it is intended to
mainly in comparative studies [5]. Even though obtain a mathematical model that can be used
the direct methods are preferred, in the case for the odour concentration in air estimation,
when these analyses are very complex, involves without direct measurement, except the initial
very high costs or impracticable conditions, phase.
EXPERIMENTAL PART
Location. The case study was conducted at a requires a 1500 m sanitary protection area. The
livestock farm operating at maximum capacity, farm is in a hilly area, in the immediate vicinity
which, according to Order no. 994/2018 [12] of a locality, characterized by winds in the
Romanian Journal of Ecology & Environmental Chemistry ● No.1 ● 2019
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4
E. Bucur et. al.: Odour pollution assessment through indirect methods based on the monitoring of technological
parameters - case study
direction of NV, as shown by the graphical expression of the wind rose (Figure 1).
The very small distance, sometimes under 100 m monitoring, iv) obtaining the equation of the
from houses, determined over the years some mathematical model and its validation by direct
problems due to the air pollution with NH3, measurements and by the mathematical
H2S, and odours largely due to the activities modelling of the odour dispersion in the air.
carried out on the farm but also to the non- Based on the analysis of technological processes
compliance with the sanitary protection area. from a livestock farm and the main BAT
Technological processes carried out on the farm techniques [13] to reduce the odour level but
meet the requirements of the BAT Conclusions also on the specific conditions of the farm in
[13], both in terms of the application of the best which the case study was conducted, three
technologies and also regarding the methods for potential sources of odour pollution and strong
reducing pollutant emissions in the air. Thus, smelling compounds have been identified:
animal feeding and shelter ventilation systems ventilation openings for animal shelters (NH3,
are automated, the ventilation rate being set odour), the slurry transfer system (NH3, H2S,
according to the temperature and humidity odour) and the lagoons for storage / treatment of
parameters of the shelter to ensure optimal slurry (NH3, H2S, smell) to which are added the
animal development conditions. The slurry is sources of diffuse emissions. In these conditions
automatically transferred to pre-sealed lagoons the parameters that can characterize the
that prevent soil and ground infiltration and are emissions of odour and strong smelling specific
covered with sealed geomembranes where they compounds, NH3 and H2S, which were taken
are enzymatically treated to reduce into account for the mathematical model
pollutant/odour emissions in the air. development are: i) ventilation rate, expressed in
Materials and Methods. The evaluation method % from the maximum capacity (vent-cap), ii)
is based on the statistical correlation and distance to shelters, m (dist), iii) distance to
multiple regression analysis applied to the data lagoons, m (dist), iv) wind direction, expressed
series obtained by monitoring the odour in degrees N (dir-vant), v) wind speed, m/s (vit-
concentration, the main specific odorous vant), vi) atmospheric temperature, 0C (temp),
pollutants (NH3 and H2S) and the technological / (vii) atmospheric humidity, % (RH) and viii) the
geographic parameters and involves the concentration of NH3 and H2S in air (in the case
following steps: i) identification the main of odour calculation) in μg/m3 or mg/m3. For
sources of odour emission and their specific concentration of NH3 and H2S determination,
parameters, ii) monitoring of the identified Horiba automatic analysers were used, with
parameters and of the odour concentration, detection limits of 5 ppbv for NH3 and 2 ppbv
iii) correlation and multiple regression analysis for H2S. The odour analyzis was done following
applied to the data series obtained by the dynamic olfactometry method according to
Romanian Journal of Ecology & Environmental Chemistry ● No.1 ● 2019
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5
E. Bucur et. al.: Odour pollution assessment through indirect methods based on the monitoring of technological
parameters - case study
SR EN 13725:2003 using a Odournet T08 correlation is always the first step in predictive
dynamic olfactometer, developed by Odournet studies preceding regression analysis.
GMBH, Germany. Meteorological parameters The regression represents the process by which
were monitored using a MetPack Wind Sonic it can estimate the value of the dependent
Portable Weather Station. For the statistical variable based on the values of the predictor
interpretation of the data - Pearson correlation variables and the mathematical relationship
and multiple regression analysis - the SPSS 20.0 expressing their interdependence. Such an
(Statistical Package for Social Sciences, version estimate is, in fact, a prediction. In this case
20.0) software was used. The correlation study, the multiple regression was used, which
analysis provides information regarding the basically relies on the same fundamental
intensity and direction of the statistical link concepts as simple linear regression, but must
between two or more variables without deal with the specific problems of a multivariate
necessarily involving a causal link between model. The multiple regression equation (Ec. 1)
them. In the case study Pearson correlation was is similar to the simple regression equation,
used, whose coefficient (r) can take values in the applied to several predictors:
range [-1, + 1]; the closer is the value of the r (1)
coefficient to ± 1, the better is the correlation
between the two analyzed parameters. To assess in which:
the statistical significance of the correlation x1…xk = values of the k predictor
coefficient and the regression coefficients, the p variables
value - the probability that the value of the a = ordinate to origin
correlation coefficient is equal to zero (the null b1…bk = regression coefficients
hypothesis) - is used; if the probability is less corresponding to the k predictor
than the significance level (ex. p<0.05 for a variables
confidence level of 95%), then the correlation Y = estimate values for the criterion variable
coefficient is statistically significant. Statistical (dependent)
0 1 2 3 4 5 6 7 8 9 10 11
6 On the road (pension) 35.60 3.98 8.00 23.00 41.00 95.00 440.00 810.00 213.00 1.35
In the farm, close to the
7 36.30 13.06 18.00 23.00 42.00 100.00 80.00 520.00 171.00 0.70
monitoring system
CMA (30 min) 15 0.300
Average 41.15 5.65 20.48 21.71 43.71 70.42 270.00 468.57 176.00 1.305
Standard deviation 4.789 1.672 5.470 .837 2.089 7.55 46.291 114.775 15.87 .245
Median 36.30 3.98 18.00 23.00 41.00 64.00 290.00 600.00 171.00 1.35
Minimum value 33.20 1.30 4.40 17.00 41.00 50.00 80.00 .00 104.00 0.60
Maximum value 69.50 13.06 47.00 23.00 56.00 100.00 440.00 810.00 220.00 2.19
8 30m from pt. 2 46.5 3.21 29 18 54 35 220 200 110 0.9
On the road, close to the
9 42.4 10.1 24 20 46 80 120 410 165 1.2
office
Colour legend:
Colour
Correlation significance very week week moderate good very good
Under these conditions, the results of the data series obtained in the first 7 measurement
Pearson correlation analysis provide the first points (Table 1), including the concentrations of
indices for predictor variables with significance NH3 and H2S; points 8 and 9 were used to
for the value of the dependent variable (odour validate the mathematical model.
concentration). Once a correlation between the The results of multiple regression (Table 3) for
variables is established, the statistical regression odour concentration as a dependent variable
method is used to establish the mathematical eliminate as statistically insignificant all
relationship. In our case, multiple regression, the variables from the equation, except the distance
Forward method, was applied to the monitoring from the lagoon, the only statistically significant
Model Summaryb
Model R R Adjusted R Std. Error of Change Statistics
Square Square the Estimate R Square F Change df1 df2 Sig. F
Change Change
1 .847a .718 .662 8.41942 .718 12.728 1 5 .016
a. Predictors: (Constant), dist-lag
b. Dependent Variable: odour
Coefficientsa
Model Unstandardized Coefficients Standardized T Sig. Collinearity Statistics
Coefficients
B Std. Error Beta Tolerance VIF
(Constant) 39.408 6.185 6.371 .001
1
dist-lag -.040 .011 -.847 -3.568 .016 1.000 1.000
a. Dependent Variable: odour
Excluded Variablesa
Model Beta In t Sig. Partial Collinearity Statistics
Correlation Tolerance VIF Minimum
Tolerance
dist .166b .631 .562 .301 .927 1.078 .927
b
dir-vant -.013 -.044 .967 -.022 .814 1.229 .814
vit-vant .064b .236 .825 .117 .953 1.049 .953
b
temp .003 .013 .990 .007 .983 1.017 .983
1
RH -.028b -.105 .922 -.052 .996 1.004 .996
vent-cap .051b .168 .875 .084 .757 1.321 .757
b
H2S -.378 -1.141 .317 -.496 .484 2.064 .484
NH3 -.181b -.725 .509 -.341 1.000 1.000 1.000
a. Dependent Variable: odour
b. Predictors in the Model: (Constant), dist-lag
The value of R2 = 0.718 indicates that this case, however, this parameter characterizes the
variable determines 71.8% of the odour H2S emission only in a proportion of 51.6%. In
concentration in the analyzed area. We mention the case of NH3 concentration, the results of the
that these results apply only to the conditions multiple regression analysis reveal two
specific to the period in which the measurements statistically significant parameters, the distance
and data sets were used. from the animal shelters and the ventilation rate,
Applying the same procedure in the case of H2S, which together account for 94.8% of the
similar results have been obtained, emission.
demonstrating that odour and H2S emissions are The equations of the mathematical models for
from the same source, i.e. the lagoon in which the three indicators and statistically significant
the slurry effluent is discharged, the only parameters can be found in Table 4.
parameter with statistical significance. In this
Table 4. Equations of mathematical models for estimating odour concentration, NH3 and H2S
Indicator Statistically significant indicators Equation of the mathematical model
Odour - distance from the lagoon, m (dist-lag)
x = distance from the lagoon, m
Y = estimated value for the odour concentration, ou e/m3
H2S - distance from the lagoon, m (dist-lag)
x = distance from the lagoon, m
Y = estimated value for the concentration of H2S, µg/m3
NH3 - ventilation rate, % from maximum
capacity (vent-cap) x1 = distance from the shelter, m
- distance from the shelters, m (dist) x2 = ventilation rate, %
Y = estimated value for the concentration NH3, mg/m3
REFERENCES
1. GREGORY, L., KENDALL, D., BARNARD, N., J Air Pollut Control Assoc, 19(2), 1969, p.
91;
2. SR EN 13725:2003 Air quality. Determination of odour concentration by dynamic
olfactometry;
3. VASILE, A., DANCIULESCU, V., TANASE, G., KIM, L., DEDIU, V., Rev. Chim.
(Bucharest), 68(8), 2017, p. 1749;
4. JIANG, J., COFFEY, P., TOOHEY, B., J Air Waste Manag Assoc, 56, 2006, p. 675;
5. HUDON, G., GUY, C., HERMIA, J., J Air Waste Manag Assoc, 50, 2000, p. 1750;
6. MORRISON, S., FORDYCE, F.M., SCOTT, E.M., Environ Geochem Health, 36, 2014, p.
319.
7. YOO, J.M., LEE, Y.R., KIM, D., JEONG, M.J., STOCKWELL, W.R., KUNDU, P.K., OH,
S.M., SHIN, D.B., LEE, S.J., Atmos Environ, 82, 2014, p. 226.
8. KLIUCININKAS, L., KRUGLY, E., STASIULAITIENE, I., RADZIUNIENE, I.,
PRASAUSKAS, T., JONUSAS, A., KAUNELIENE, V., MARTUZEVICIUS, D., Atmos
Environ, 97, 2014, p. 83.
9. BUCUR E., DANET, A.F., LEHR, C.B., LEHR, E., NITA-LAZAR, M., J Air Waste Manag
Assoc., 67(4), 2017, p. 391.
10. DANCIULESCU, V., BUCUR, E., PASCU, L.F., VASILE, A., BRATU, M., J Environ Prot
Ecol., 16(3), 2015, p. 815
11. http://eippcb.jrc.ec.europa.eu/reference/ - JRC Reference Report on Monitoring of
Emissions to Air and Water from IED installations, 2017.
12. Ordinul nr. 994/2018 pentru modificarea si completarea Normelor de igiena si sanatate
publica privind mediul de viata al populatiei, aprobate prin Ordinul ministerului sanatatii nr.
119/2014.
13. Commission Implementing Decision (EU) 2017/302 of 15 February 2017 establishing best
available techniques (BAT) conclusions, under Directive 2010/75/EU of the European
Parliament and of the Council, for the intensive rearing of poultry or pigs.
14. STAS 12574-87. Aer din zonele protejate. Conditii de calitate (in Romanian).
Degradation experiments on 5-Fluorouracil synthetic solutions were performed using three types of
advanced oxidation systems: UV/H2O2, UV/TiO2 and UV/H2O2/TiO2. Optimum parameters for all
three systems were established and 5-Fluorouracil degradation efficiencies were calculated, for all
systems being more than 97%. The results showed that the combined UV/H2O2/TiO2 system is
offering shortest irradiation time, the possibility to recover and reuse the photo catalyst as well as
the possibility to use solar radiation. Obtained results proved also that advanced oxidation
processes represent a viable option for degradation of hazardous pollutants that cannot be removed
properly via conventional wastewater treatment processes.
Keywords: 5-Fluorouracil, TiO2, H2O2, degradation efficiency
5-Fluorouracil (5FU) is one of the most common drugs used in cancer treatment worldwide [1].
H
N O
NH
F
O
Fig. 1. 5-Fluorouracil (5FU) structure
Due to its non-selective action it affects not only concentrations within the sewerage system of
cancerous but healthy cells, presenting an hospitals are low,varying from micrograms to
carcinogenic, genotoxic, mutagenic and hundreds of micrograms, its removal through
teratogenic potential [2]. Moreover about 15% conventional wastewater treatment processes is
of the administered 5FU is leaving the human not possible asking for application of more
body with no transformation ending within the performant processes [5], [6]. Therefore, 5FU
sewerage network [3]. Recent studies degradation experiments were performed in
demonstrated that this type of pollutant cannot order to determine the optimum parameters for
be removed within the wastewater treatment the following systems: UV/H2O2, UV/TiO2 and
plants [4]. Even if cytostatic residues UV/H2O2/TiO2.
EXPERIMENTAL PART
Degradation experiments were performed using Agilent 240MS ionic trap spectrometer [7].
a Heraeus UV reactor with the following Synthetic solutions were prepared using 5FU
characteristics: UV lamp - TQ150-Z3 type, supplied from Sigma-Aldrich.
cooling jacket, reaction vessel from quartz, All samples were bubbled with air (50 L/h) prior
volume = 400 cm3, equipped with a magnetic to irradiation in order to avoid electrons – holes
recirculation pump. Determination of 5FU recombination.
concentration was carried out using an Agilent All experiments were at least duplicated in order
7890A gas chromatograph coupled with an to assure their repeatability.
Fig. 3. H2O2 dose effect upon the 5FU degradation in UV/H2O2 system
[5FU]0 = 13.35 mg/L=1.03 x 10-4 M; irradiation time = 30 min
Results showed that optimum H2O2 dose [5FU]0 = 13.35 mg/L = 1.03 x 10-4 M for H2O2
correspond to [5FU]0/[H2O2] = 1:17 which dose corresponding to [5FU]0/[H2O2] = 1:17,
assures a degradation efficiency of 91.69% after and irradiation time up to 45 minutes.
30 minutes of irradiation. Optimum dose was Experimental results showed that the increase of
used in all the subsequent experiments for irradiation time has a positive impact upon 5FU
UV/H2O2 system. degradation efficiency. Therefore, the optimum
Effect of irradiation time irradiation time was set at 45 minutes, which
In order to establish the optimum irradiation assured the substrate degradation with a 97.08%
time 5FU degradation experiments were efficiency and a residual 5FU concentration of
performed for an initial 5FU concentration 0.39 mg/L.
UV/TiO2 system
Previous reported researches on 5FU 13.35 mg/L = 1.03 x 10-4 M; photo catalyst
degradation in UV/TiO2 system [7] led to the concentration [TiO2] = 100 mg/L at pH = 7
following optimal parameters: pH = 7; photo (optimum conditions for UV/TiO2 system).
catalyst dose 100 mg/L; irradiation time 180 Effect of H2O2 dose
minutes. These conditions assured 5FU The oxygen presence plays an important role
degradation with an efficiency of more than due to its electron scavenger properties fact that
99%. Moreover, the processes obey Langmuir – is affecting electrons – holes recombination
Hinshelwood model proving that degradation process. It was demonstrated that oxygen can
process occurs mainly on TiO2 particle surface. have both a positive or negative impact upon the
UV/H2O2/TiO2 system pollutant degradation [8].
From the main parameters that are influencing In order to investigate these parameters; molar
the degradation processes the following were ratios [5FU]0/[H2O2] were varied within the
investigated: H2O2 dose and irradiation time. All domain 1:0.5 – 1:5. Obtained experimental
experiments were performed using a synthetic results are presented within Table 2 and Figure
solution with initial concentration [5FU]0 = 5.
Obtained results proved that optimum H2O2 dose The increase of irradiation time has a positive
corresponds to the [5FU]0/[H2O2] ratio of 1:5. effect upon 5FU degradation efficiency.
Effect of irradiation time Optimum irradiation time was set at 45 minutes,
For the optimum H2O2 dose, the influence of assuring the substrate degradation with a
irradiation time upon 5FU degradation 97.37% efficiency and a residual 5FU
efficiency was investigated, the results being concentration of 0.139 mg/L.
presented within Figure 6.
CONCLUSIONS
Optimum parameters for 5FU degradation using of using solar radiation simultaneously with the
UV/H2O2, UV/TiO2, UV/H2O2/TiO2 were shortest irradiation time that leads to energy
established and degradation efficiencies for all costs reduction and also to reduction of
three systems were calculated. implementation (compact installations) and
A synthesis of these data is presented within operating costs. It also offers the possibility to
Table 3. recover the photo catalyst and reuse it in further
UV/H2O2/TiO2 system presents the advantage treatment cycles.
REFERENCES
1. LIN, H., LIN, A., Water Res., 48, 2014, p. 559.
2. KUMMERER, K., HAIB, A., SCHUSTER, A., HEIN, A., EBERT, I., Environ. Sci. Pollut.
Res. Int, 23, no. 15, 2014, p. 14791.
3. ZHANG, J., CHANG, V., GIANNIS, A., WANG, J., Sci. Total Environ., 445, 2013, p. 281.
4. WANG, X.-H., LIN, A.-C., Environ. Pollut., 186, 2014, p. 203.
5. CONSTANTIN, L., NITOI, I., CRISTEA, I., OANCEA, P., ORBECI, C., NECHIFOR
A.C., Rev. Chim. (Bucharest), 66, no. 5, 2015, p. 597.
6. NITOI, I., OANCEA, P., CRISTEA, I., CONSTANTIN, L., NECHIFOR, G., J. Photochem.
Photobio. A, 298, 2015, p. 17-23.
7. CONSTANTIN, L., NITOI, I., CRISTEA, I., OANCEA, P., Rev. Chim. (Bucharest), 67,
no. 8, 2016, p. 1447.
8. MOZIA, S., Sep. Purif. Technol., 73, 2010, p. 71
TOMA GALAON, ALINA BANCIU, FLORENTINA LAURA CHIRIAC AND MIHAI NITA-
LAZAR*
National Research and Development Institute for Industrial Ecology ECOIND Bucharest, 71-73 Drumul Podul Dambovitei, 060652,
Bucharest, Romania
*corresponding author (e-mail): [email protected]
The worldwide population rise corroborated with the raise of the health-care standards have
generated an escalation of the antibiotic production and uncontrolled usage. The subsequent effects
of this escalation have led to an increase of the antibiotic resistance rates, Romania is in the top of
the EU countries regarding the antibiotic resistance rates, and to a continuous presence in the
environment, including the aquatic environment. Unfortunately, the present design of the classical
WWTPs is not optimized for the efficient removal of antibiotics since these compounds may have
highly soluble and polar molecular structures. Instead, antibiotics removal using microorganisms
could be an ecofriendly solution to this environmental issue, as long as their antibiotic degradation
structures are not more toxic than the antibiotic itself.
In the present review, we focus on the environmental presence and biodegradation of the most
commonly used antibiotics as well as on their biodegradation, based on bacterial model, monitored
by mass-spectrometric methods.
The penicillin discovery by Fleming in 1928 effect was effectively proved by E. Duchesne in
was a turning point in the medicine and 1897. First reports on penicillin were in 1938,
pharmacology fields, but the quest of finding but soon after that, 1939-1940, S. Waksman and
chemical compounds with microbial growth R. Dubos reported over 20 potential antibiotics.
inhibition activity started long time before. Since then there is a continuous race between
In 1871 J. B. Sanderson and his coworkers antibiotic resistance and synthesis of new
reported a mold strain, Penicillium, with chemical compound with antibacterial effect.
bacterial growth inhibition effect, but the actual
At the present, a growing number of chemical brief description of some of these classes of
compounds with more and more specific antibiotics is presented in the following
antimicrobial activity, antibiotics, have been paragraphs together with their structural
continuously synthetized for human, animal and characteristics which are given in Figure 2.
plants health. There are many classes of Beta-lactams are one of the most used
antibiotics regardless of their natural, synthetic classes of antibiotics containing a beta-
and semi-synthetic origin [2] with a very well lactam ring which consists of a 3-carbon
characterized antibacterial effect (Table I) [3] and 1-nitrogen atom ring and a ketone
and subsequently there are largely used due to group.
their benefic effects, up to 2,000,000 tons of These antibiotics interfere with proteins
antibiotics (AB) consumption per year, essential for synthesis of bacterial cell wall
according to Centers for Disease Control and [4]. Some of the most prescribed
Prevention in 2015. representatives are Penicillins (Penicillin G,
Antibiotics structural characterization Penicillin V, Oxacillin, Ampicillin,
From a structural point of view antibiotics Amoxicillin), Cephalosporines,
can be divided into several types of Monobactams (Aztreonam), and
chemical structures. Logically, this Carbapenems (Imipenem, Meropenem,
classification is co-related with their Ertapenem).
mechanism of action (bactericidal, Macrolides are large molecules containing
bacteriostatic, bacteriolytic) and their a 14 to 16 membered macrolide ring
spectrum of activity (types of controlled (macrocyclic lactose ring). They inhibit
bacteria). The most important classes of protein synthesis by bacteria leading
antibiotics are summarized in Table 1. A sometimes to cell death. Some of the most
E. coli, Salmonella, Chlamydia trachomatis In all those areas, especially wastewaters and
[9]. Main members of this class are: WWTP, there have been a mixt of antibiotic
Sulfamethoxazole, Sulfadiazine, chemical structures as well as an exchange
Sulfamethazine, Sulfadimethoxine, between human and environmental bacteria
Sulfanilamide, Sulfoxazole, etc. From could generate environmental stress and
structural point of view, Sulfonamides have antibiotic resistance [24, 25]. The antibiotic
a common sulfonamide group grafted on resistance has been accentuated by the
different hydrocarbon structures (aliphatic persistence of low molecular weight (< 1000 D)
or aromatic in nature). These antibiotics are antibiotics which are very soluble in water
given to treat urinary tract infections, bodies such as b-lactams, aminoglycosides,
septicemia, meningitis and so on. lincosamides, macrolides, nitrofurans,
Antibiotic resistance genes acquisition and amphenicols, quinolones, fluoroquinolones,
dissemination rifamycins, sulfonamides and tetracyclines [15,
In addition to the human consumption of 26]. The wastewater treatment has not been
antibiotics, they are widely used in agriculture sufficient to completely eliminate
enhancing the livestock growth, bee-keeping, pharmaceutical compounds, including ABs [17]
and fish farming [10]. Unfortunately, the and the lower remaining amount released to the
antibiotic low degree of metabolization and its environment boost the antibiotic resistance
high environmental reminiscence combined with process.
a high dissipation property has become a major Long-term exposure at lower concentration of
threat to public health by compromising antibiotics than their minimal inhibitory
bacterial infection treatments. In spite of concentration (MIC) increased the bacterial
banning the usage of antibiotics in agriculture DNA mutation rate of genes, the horizontal gene
and livestock industries in EU from 2006, a transfer (HGT) in the bacterial population and
large number of world population from China modulate genes expression, altogether inducing
and India keep using them in agriculture and an antibiotic resistance [27, 28, 29] as well as
livestock industries [11]. further increasing the minimal inhibitory
Moreover, the excessive use of antibiotics in concentration (MIC) of antibiotics [30].
human and animal medicine [12] over amplify Antibiotic resistance mechanisms
the antibiotic presence in the environment and The ARGs allowed (micro)organisms to develop
more dangerously this has triggered the selection resistance by avoiding a specific drug-target
of antibiotic resistant bacteria (ARB) and their interaction such as DNA, RNA or protein,
antibiotic resistance genes (ARGs) [13]. The use rejecting of antibiotics from the cell through
of antibiotics in hospitals did not mean that efflux pump (Fig. 3a) and antibiotics
humans are their only target and, unfortunately, degradation (Fig. 3b) by biotic and abiotic
the antibiotics have had also an impact on others processes [3].
not-targeted organisms [14, 15] inducing The functionality of ARGs is extended also to
resistance in all organisms after a longtime the survival fight between bacterian strains
exposure [16]. Significant quantities (30–90%) which naturally produce antibiotics to eliminate
of consumed antibiotics may be eliminated the competition for the same resources. The
unchanged or as active metabolites through structural characteristics of cell walls (Gram
urinary and/or fecal excretion [17, 18]. In spite positive vs Gram negative bacteria –extra outer
of the fact, that hospitals from European membrane- or exopolysaccharide and
countries discharge only 5–20% extracellular DNA) as well as the bacterial
pharmaceuticals, the antibodies concentrations aggregation in biofilms [31] prevent antibiotic to
in hospital effluents have been 100-fold higher reach their targets increasing the level of
than those of municipal effluents [19, 20]. The resistance [32, 33]. Moreover, the biofilm is
antibiotics could be detected in municipal suitable environment for resistance genes
sewage water, WWTP biosolids, effluents, soil, acquisition and spreading among bacterial
surface waters, groundwaters, sediments, biota populations [34].
and drinking water [21, 22, 23].
Tetracyclines have smaller structure than others. The main mechanism of action is by
cephalexin and degrade faster by UV-light, inhibiting DNA replication. For example,
and photolysis [42]. Overall tetracyclines, Ciprofloxacin and Ofloxacin inhibit DNA
sulphonamides, tylosin, nitrofuran or topoisomerases of bacteria, affecting thus
(fluoro)quinolones antibiotics are sensitive nucleic acid synthesis. In a biodegradability
to UV light, but fluoroquinolones are study [58] the authors presented several possible
resistant to hydrolysis [43]. Oxygen and bacteria biodegradation pathways (LC-MS/MS
ozone are widely spread abiotic factors and identification) of Ciprofloxacin generated by a
they are involved in the sulfamethoxazole mixed culture of Gammaproteobacteria,
and oxytetracycline antibiotics degradation Bacteroidia, and Betaproteobacteria. Authors
by oxidation [44, 45]. Metal ions such identified several biodegradation products of
mercury, copper, zinc, cadmium, and cobalt Ciprofloxacin generated by deamination (C-N
are involved in beta-lactam antibiotics bond cleavage), hydroxylation, defluorination
degradation, like Cephalosporin and (C-F bond cleavage), dealkylation (C-C bond
Penicillin, by breakage of the β-lactam ring cleavage) reactions. The structural changes of
[46]. Ciprofloxacin are promoted by bacteria
Enzymatic degradation of antibiotics enzymatic degradation. Approximately 47% of
The enzymes play an important part in the the Ciprofloxacin amount was biodegraded by
mechanism of antibacterial resistance by bacteria after 7 days and more than 89% the
modifying or degrading the antibiotics (Fig. 3b). antibiotic was dissipated after 28 days of
Studies showed a correlation between ARG incubation.
expression, antibiotic degrading enzyme The Sulfonamide class of antibiotic drugs are
production and antibiotic resistant bacteria. In another important group of antibiotics with high
addition, some studies point to the idea that volume of consumption worldwide. Among its
ARGs involved in the expression of representatives, some of the most important are
aminoglycoside-modifying and β-lactamase represented by Sulfamethoxazole (SMX),
enzymes were present in the environment long Sulfadiazine (SDZ), Sulfamethazine (SMZ),
before the use manmade antibiotics in a Sulfadimethoxine (SDM), Sulfanilamide (SAD)
therapeutic scope [47, 48, 49] showed that β- and others. In a comprehensive review, Chen et
lactamase family shared a common origin with al., describe the recent advances in
enzymes involved in cell wall biosynthesis. understanding Sulfonamide antibiotics
In spite of using the antibiotics and their biodegradation by microorganisms present in
presence in the environment, bacteria resistant natural and engineered active sludge systems
to β-lactams are thriving and there are a [9]. The authors describe biodegradation
constant presence in more and more countries pathway of Sulfonamides which is mainly
[50, 51, 52]. generated by hydroxylation and acetylation
The same emerging resistance pattern it is also reactions at the aniline or the aminated
observed for bacteria producing carbapenemase heteroaromatic groups of the sulfonamide
[53, 54, 55]. Unfortunately, bacteria producing structure. Other chemical changes arise by C-N,
carbapenemase, cephalosporinase or oxacillinase C-S and C-C bond cleavage, along with
exchange their ARG among them or with other oxidation. As with respect to the main bacterial
bacteria from the environment, subsequently population able to degrade these types of
spreading the resistance to antibiotics to other compounds, the authors report Proteobacteria
bacterial communities and other distant and Acidobacteria, Firmicutes and
geographical areas [56, 57]. Fluoroquinolone Bacteroidetes for Sulfamethazine (SMZ)
class of antibiotics is one of the most prescribed biodegradation [59]. In the case of
group of antibiotics worldwide. They are used (Sulfanilamide) SAD biodegradation was
for treating infections generated by aerobic promoted by Firmicutes, Bacteroidetes, Bacillus
gram-positive and gram-negative bacteria. and Chryseobacterium genus groups [60].
Among fluoroquinolones, some of the most Concerning activated sludge bacteria species
important representatives are Ciprofloxacin, which are able to biodegrade many of the
Norfloxacin, Levofloxacin, Ofloxacin and sulfonamide antibiotics, it is worthwhile to
REFERENCES
1. VENTOLA, C.L., The Antibiotic Resistance Crisis, Part 1: Causes and Threats, P T. 2015,
40, no. 4, p. 277–283.
2. CATTEAU, L., ZHU, L., Van BAMBEKE, F., QUETIN-LECLERCQ, J., 2018.
Phytochem. Rev., p. 1–35.
3. KUMAR, V., BAWEJA, M., LIU, H., SHUKLA, P., 2017. Springer, Singapore, p. 259–273.
4. HEESEMANN, J., 1993. Infection., 21, no. 1, p. 4-9.
5. ETEBU E., ARIKEKPAR, I., 2016. Int. J. Appl. Microbiol. Biotehnol. Res., 4, p. 90-101.
6. CHOPRA, I., ROBERTS, M., 2001. Microbiol. Mol. Biol. Rev., 65, no. 2, p. 232-260.
7. DOMAGALA, J. M., 1994. J. Antimicrob. Chemother, 33, p. 685-706.
8. PETERSON, L. R., 2008. Clin Microbial. Infect., 14, no. 6, p. 30-45.
9. CHEN, J., XIE, S., 2018. Sci. Tot. Environ., 640–641, p. 1465–1477.
10. HONG, B., LIN, Q., YU, S., CHEN, Y., CHEN, Y., CHIANG, P., 2018. Sci. Total Environ.,
634, p. 448–458.
11. RONQUILLO, M.G., HERNANDEZ, J.C.A., 2017. Food Control, 72, p. 255–267.
12. PRUDEN, A., LARSSON, D.G.J., AMÉZQUITA, A., COLLIGNON, P., BRANDT, K.K.,
GRAHAM, D.W., LAZORCHAK, J.M., SUZUKI, S., SILLEY, P., SNAPE, J.R., TOPP, E.,
ZHANG, T., ZHU, Y.-G., 2013. Environ. Health Perspect., 121, p. 878–885.
13. HUGHES, D., 2014. IUBMB Life, 66, p. 521–529.
14. LEUNG, H.W., MINH, T.B., MURPHY, M.B., LAM, J.C., SO, M.K., MARTIN, M.,
RICHARDSON, B.J., 2012. Environ. Int., 42, p. 1–9.
15. GRENNI, P., ANCONA, V., CARACCIOLO, A.B., 2018. Microchem. J., 136, p. 25–39.
16. TACCONELLI, E., CARRARA, E., SAVOLDI, A., HARBARTH, S., MENDELSON, M.,
MONNET, D.L., OUELLETTE, M., 2018. Lancet Infect. Dis., 18 no. 3, p. 318–327.
17. TIWARI, B., SELLAMUTHU, B., OUARDA, Y., DROGUI, P., TYAGI, R.D., BUELNA,
G., 2017. Bioresour. Technol., 224, p. 1–12.
18. SHAO, S., HU, Y., CHENG, J., CHEN, Y., 2018. Crit. Rev. Biotechnol., 38, p. 1195–1208.
19. CARRARO, E., BONETTA, Si, BERTINO, C., LORENZI, E., BONETTA, Sa, GILLI, G.,
2016. J. Environ. Manag., 168, p. 185–199.
20. SANTOS, L.H.M.L.M., GROS, M., RODRIGUEZ-MOZAZ, S., DELERUE-MATOS, C.,
PENA, A., BARCELÓ, D., MONTENEGRO, M.C.B.S.M., 2013. Sci. Total Environ., 461–
462, p. 302–316.
21. BARANCHESHME, F., MUNIR, M., 2018. Front. Microbiol., 8, p. 2603.
22. ZHANG, H., JIA, Y., KHANAL, S.K., LU, H., FANG, H., ZHAO, Q., 2018. Environ. Sci.
Technol., 52, no. 11, p. 6476–6486.
23. WILLIAMS, M., KOOKANA, R.S., 2018. Health Care Environ. Contam., 11, p. 21.
24. AMOS, G.C.A., PLOUMAKIS, S., ZHANG, L., HAWKEY, P.M., GAZE, W.H.,
WELLINGTON, E.M.H., 2018. ISME J., 12, p. 681–691.
25. HOCQUET, D., MULLER, A., BERTRAND, X., 2016. J. Hosp. Infect., 93, p. 395–402.
26. KRZEMINSKI, P., TOMEI, M.C., KARAOLIA, P., LANGENHOFF, A., ALMEIDA,
C.M.R., FELIS, E., RIZZO, L., 2018. Sci. Total Environ., 648, p. 1052–1081.
27. GULLBERG, E., CAO, S., BERG, O.G., ILBÄCK, C., SANDEGREN, L., HUGHES, D.,
ANDERSSON, D.I., 2011. PLoS Pathog., 7, e1002158.
28. PARTRIDGE, S.R., KWONG, S.M., FIRTH, N., JENSEN, S.O., 2018. Clin. Microbiol.
Rev., 31.
29. WISTRAND-YUEN, E., KNOPP, M., HJORT, K., KOSKINIEMI, S., BERG, O.G.,
ANDERSSON, D.I., 2018. Nat. Commun., 9, no. 1, p. 1599.
30. TRINH, T.D., ZASOWSKI, E.J., CLAEYS, K.C., CASAPAO, A.M., COMPTON, M.,
LAGNF, A., RYBAK, M.J., 2018. Infect. Dis. Ther., 7, no. 1, p. 161–169
31. QI, L., LI, H., ZHANG, C., LIANG, B., LI, J., WANG, L., DU, X., LIU, X., QIU, S.,
SONG, H., 2016. Front. Microbiol., 7, p. 483.
32. MAURICE, N.M., BEDI, B., SADIKOT, R.T., 2018. Am. J. Respir. Cell Mol. Biol., 58, no.
4, p. 428–439.
33. AHMED, M.N., Porse, A., SOMMER, M.O.A., HOIBY, N., CIOFU, O., 2018. Antimicrob.
Agents Chemother, 62, no.8, e00320–18.
34. FUX, C.A., COSTERTON, J.W., STEWART, P.S., STOODLEY, P., 2005. Trends
Microbiol., 13, p. 34–40.
35. PETCHIAPPAN, A., CHATTERJI, D., ACS Omega 2017, 2, p. 7400-7409
36. ENDICOTT J.A., LING V., 1989. Annual Rev Biochem., 58, p. 137-171.
37. VAN VEEN H.W., CALLAGHAN R., SOCENEANTU L., SARDINI A, KONINGS W.N.,
HIGGINS CF. (1998). Nature, 391, p. 291-5.
38. LI, B., ZHANG, T., 2010. Environ. Sci. Technol., 44, p. 3468–3473.
39. MASSÉ, D.I., SAADY, N.M.C., GILBERT, Y., 2014. Animals, 4, no. 2, p. 146–163.
40. AHN, Y., Jung, J.Y., VEACH, B.T., KHARE, S., GOKULAN, K., PIÑEIRO, S.A.,
CERNIGLIA, C.E. Regul. Toxicol. Pharmacol. 99, p. 105–115.
41. ZHANG, R., YANG, Y., HUANG, C.H., ZHAO, L., SUN, P., 2016. Water Res., 103, p.
283–292.
42. AZIMI, S., NEZAMZADEH-EJHIEH, A., 2015. Mol. Catal. A Chem., 408, p. 152–160.
43. OKAIKUE-WOODI, F.E., KELCH, S.E., SCHMIDT, M.P., MARTINEZ, C.E.,
YOUNGMAN, R.E., ARISTILDE, L., 2018. J. Colloid Interface Sci., 513, p. 367–378.
44. SIRÉS, I., BRILLAS, E., 2012. Environ. Int., 40, p. 212–229.
45. BENNER, J., SALHI, E., TERNES, T., VON GUNTEN, U., 2008. Water Res., 42, no. 12,
p. 3003–3012.
46. BISCHOFF, S., WALTER, T., GERIGK, M., EBERT, M., VOGELMANN, R., 2018. BMC
Infect. Dis., 18, no. 1, p. 56.
47. D'COSTA, V.M., KING, C.E., KALAN, L., MORAR, M., SUNG, W.W.L., SCHWARZ,
C., FROESE, D., ZAZULA, G., CALMELS, F., DEBRUYNE, R., GOLDING, G.B.,
POINAR, H.N., WRIGHT, G.D., 2011. Nature, 477, p. 457–461.
48. SONG, J.S., JEON, J.H., LEE, J.H., JEONG, S.H., JEONG, B.C., KIM, S.-J., LEE, J.-H.,
LEE, S.H., 2005. J. Microbiol., 43, p. 172–178.
49. MEROUEH, S.O., MINASOV, G., LEE, W., SHOICHET, B.K., MOBASHERY, S., 2003.
J. Am. Chem. Soc., 125, p. 9612–9618.
50. BAJAJ, P., KANAUJIA, P.K., SINGH, N.S., SHARMA, S., KUMAR, S., VIRDI, J.S.,
2016. Environ. Sci. Pollut. Res. Int., 23, p. 1954–1959.
51. LU, S.-Y., ZHANG, Y.-L., GENG, S.-N., LI, T.-Y., YE, Z.-M., ZHANG, D.-S., ZOU, F.,
ZHOU, H.-W., 2010. Appl. Environ. Microbiol., 76, p. 5972–5976.
52. TISSERA, S., LEE, S.M., 2013. Malays J. Med. Sci., 20, p. 14–22.
53. ALMAKKI, A., MAURE, A., PANTEL, A., ROMANO-BERTRAND, S., MASNOU, A.,
MARCHANDIN, H., JUMAS-BILAK, E., LICZNAR-FAJARDO, P., 2017. Int. J.
Antimicrob. Agents, 50, p. 123–124.
54. SEKIZUKA, T., YATSU, K., INAMINE, Y., SEGAWA, T., NISHIO, M., KISHI, N.,
KURODA, M., 2018.Sphere 3.
55. XU, H., WANG, X., YU, X., ZHANG, J., GUO, L., HUANG, C., JIANG, X., LI, X.,
FENG, Y., ZHENG, B., 2018. Environ. Pollut., 235, p. 931–937.
56. HENRIQUES, I.S., FONSECA, F., ALVES, A., SAAVEDRA, M.J., CORREIA, A., 2006.
Res. Microbiol., 157, p. 938–947.
57. ALLEN, H.K., DONATO, J., WANG, H.H., CLOUD-HANSEN, K.A., DAVIES, J.,
HANDELSMAN, J., 2010. Nat. Rev. Microbiol. 8, p. 251–259.
58. LIAO, X., LI, B., ZOU, R., DAI, Y., XIE, S., YUAN, B., 2016. Environ. Sci. Pollut. Res.,
23, p. 7911–7918.
59. LIAO, X.B., LI, B.X., ZOU, R.S., XIE, S.G., YUAN, B.L., 2016b. Appl. Microbiol.
Biotechnol., 100, p. 2439–2447.
60. ISLAS-ESPINOZA, M., REID, B.J., WEXLER, M., BOND, P.L., 2012. Microb. Ecol., 64,
p. 140–151.
61. YAN, N., XIA, S.Q., XU, L.K., ZHU, J., ZHANG, Y.M., RITTMANN, B.E., 2012. Appl.
Microbiol. Biotechnol., 94, p. 527–535.
The market expansion due to globalization generated more anthropogenic contaminants, enhancing
their negative impact on the environment. In order to prevent and balanced their negative effect,
more rapid, sensitive and eco-friendly ecotoxicological studies as risk assessments were developed.
Each country developed national programs to ensure the research infrastructure for
implementation and development of new testing methodologies, generating new experimental data
transferable to governmental authorities, economic agents and academia.
In this review, we intend to highlighted the progress of Romanian researchers in the last 10 years in
the field of ecotoxicology and aquatic risk assessment. The studies focused on aquatic systems as a
non-targeted system to the anthropogenic pollutants impact.
The implemented methodologies according with international requirements for chemical control
like REACH Regulation, Detergents Regulation, Biocide Products Regulation or the Aquatic Risk
Assessment Guidelines were presented.
The ecotoxicological effects of various contaminants such as detergents, pharmaceuticals,
pesticides, biocides or heavy metals and environmental polluted samples (surface water, sediment
or sludge) were provided.
Overall, the studies presented in our studies have a nationally novelty through the variety of test
substances, the complexity of laboratory experiments and data processing. The research work has a
continue applicability in industry and for control authorities. New accredited testing services that
providing useful information about the chemicals and their impact on the environmental were
introduced. In addition, the ecotoxicology studies and their challenges were a great source for
professional training like PhD and master studies and for impact published papers.
The interest of scientists to identify, solve and discipline base on principles of toxicology,
prevent the environmental problems has become biology, chemistry and ecology, being focused
a priority in the context of population growth, on the research of chemical compounds effects
waters contamination, global warming and on living organisms, other than man. The
ecosystems damage or their extinction [1]. ecotoxicology is separated in two section:
Moreover, the human well-being led to chemical pollutant dynamics – that evaluate the mobility,
market expansion that have generated new migration, transformation, degradation in
anthropogenic contaminants in larger amounts, different environmental area; and bio toxicology
thus enhancing their negative impact on the - that examine the effects of toxic compounds on
environment. The ecotoxicology of sensitive biological systems correlated with field
anthropogenic contaminants became one of the data [1].
most published life science topic, due to vast This review intends to highlight the progress of
and complex research sources. Romanian research, especially from National
Ecotoxicology field is a complex scientific Research and Development Institute for
Industrial Ecology - ECOIND Bucharest, in the more than 8500 studies were obtained based on
last 10 years in the field of anthropogenic a “ecotoxicology” topic search in Science Direct
pollutants ecotoxicology and their aquatic risk Platform. The scientists interest is focused booth
assessment. on fundamental and applicative researches. The
Trend of ecotoxicity studies fallowed issues are covered: to testing new
The ecotoxicity studies were mainly aimed the toxicants, to develop new eco-friendly biotests,
environmental protection through the to find new acceptable environmentally
compliance of international norms by chemical technologies applied in chemical industry,
industry. The studies have novelty through the agriculture and households and to elaborate new
variety of the test substances available on the environmental legislation / guidelines /
national market (such as pesticides, surfactants, regulations.
pharmaceutical compounds, biocides), the Concerning the studied ecosystems, the most
complex laboratory experiments (acute, sub- papers were published for freshwater toxicity,
acute or chronic exposure tests using different followed by marine and terrestrial studies. The
organisms and high performance chemical most used biological systems were crustaceans
detection new methods) and data analyses (such and fish followed by insects, mollusks, algae,
as risk characterization, REACH classification, bacteria and plants. The mono-species tests were
threshold approach system or natural water the most used in research studies during 2010 -
classification after Persoone 2003). 2016 followed by multispecies test and two-
The research studies performed in the last 10 species test and in finally community-cosm [1].
years match the international rising trend of This was happening because single species tests
scientific researches in Thomson ISI Web of were in the routine of laboratories, easy to be
Knowledge – Web of Science Platform (Fig. 1). reproduced and more economic.
An exponential increase of published studies in The use of bio-indicators / biomarkers in
the field of chemicals ecotoxicology was ecotoxicology is a top subject in fundamental
observed. researches in order to prevent, control and solve
The same trend is maintained also for biotests different environmental pollution issues. A
studies or risk assessments studies (Fig. 2). The search of the topic “environmental biomarkers”
search of “ecotoxicity” topic in Wiley Online or “environmental bio-indicators” on Wiley
Library revealed more than 2300 papers Online Platform reveled more than 45 000
published in the same period. The high interest published documents in the last 10 years, their
for environmental ecotoxicity studies have been number maintaining constant for each year, that
also maintained in 2019. At the start of 2019, suggest a great interest for this subject.
Fig. 1. Trend of ecotoxicological studies published Fig. 2. Trend of aquatic risk assessment studies
on Web of Science Platform published on Web of Science Platform
ecotoxicological assessment and aquatic risk European continental waters in order to protect
assessment, laboratory tests on fish, aquatic the water resources quality. The scientific
invertebrates (Daphnia magna), green algae / community as well as the EU composed a list of
aquatic plants are mostly recommended; in priority substances including harmful chemicals
addition, studies of bio-concentration and such as: pesticides, metals, volatile organic
behavior in the aquatic environment (water and compound, dioxins, polychlorinated biphenyl
sediment) - degradation, biodegradation, compound and recently pharmaceuticals
absorption, etc. are required. substances, which is regularly upgraded under
Today, Romania should respond to all European Water Framework Directive (WFD
reglementation concerning the chemicals testing 2000/60/EC). The Water Directive promote an
according with the OECD Guidelines. The integrate approach concerning the evaluation of
OECD methods on chemical testing summing natural receptors through collection and
up more than 150 standards periodically correlation of interdisciplinary data (physical
upgraded according to scientific progress, used and chemical parameters, as well as biological,
by authorities, industry or testing laboratories ecological, hydro-geomorphological and socio-
for chemicals ecotoxicological assessment. economic data).
Moreover, our interest section “Effects on Biotic Methodologies and testing strategies
Systems” include more than 40 biotests with Romania developed national programs to ensure
different species (http://www.oecd.org). the research infrastructure for implementation
The European chemicals safety guidelines and development of new testing methodologies
promote the reduction of animal (especially to generate new experimental data transferable
vertebrate) testing by introducing the mandatory to governmental authorities, economic agents
data sharing requirement and by encouraging the and academia.
use of alternative testing methods [4]. In the field of ecotoxicology were developed
Microbiotests resulted based on OECD test or adequate procedures in order to evaluate and
ISO / EPA standards as alternative tests more control the effect of anthropogenic
easy to use, economic and reproducible, ready to contaminants, more rapid, sensitive and eco-
use for establish the harmful effects of friendly bioassays as well as risk assessments
chemicals/contaminated environmental samples studies [11, 12]. For the ecotoxicity bioassay
on small organisms (invertebrates, algae, were needed different species easy to be
ciliates, rotifers, plants and bacteria) [5, 6, 7]. acclimatized in laboratory conditions, low-cost,
The aquatic risk assessment is in force since sensitive to the toxicants, internationally
2003 based on “Technical Guidance Document recognition, representative for the exposed
on Risk Assessment: Commission Directive biological system, indigenous species and
93/67/EEC on Risk Assessment for New Notified related to possible changes in the affected
Substances” publication. ecosystems. Given this, our experimental studies
Furthermore, the European Medicines were conducted on various species to cover the
Evaluation Agency (EMEA) and the US Food entire food chain (table 1). Two species of
and Drug Administration (FDA), developed and freshwater fish with economic interest for
implemented various environmental risk Romania (Cyprinus carpio and Carassius
assessment guidelines for the ecosystems safety auratus gibelio), two species of crustaceans
[8, 9,10]. The documents establish the steps for (planktonic Daphnia magna and benthic
hazard characterization through estimation of Heterocypris incongruens), one species of green
PEC (Predicted Environmental Concentration) algae (Selenastrum capricornutum renamed
and PNEC (Predicted Non-Effect Concentration Pseudokirchneriella subcapita), one rotifer
on living organisms) values. The value of PEC/ (Brachiounus calyciflorus), one protozoan
PNEC ratio is widely accepted as endpoint in (Tetrahymena thermophile), a variety of bacteria
aquatic risk assessment model taking into species, an aquatic plants (Spirodela polyrhiza)
account the unfavorable scenarios [11]. and three terrestrial plants (Lepidium sativum,
Supplementary, the EU Water Framework Sinapis alba, Sorghum saccharatum) were
Directive 2000/60/EC (WFD), suggests that the selected to be used in ecotoxicity tests. The
hazardous chemicals need to be monitored in mortality / immobilization, growth inhibitions or
biochemical changes (gene expression) were the exposure of testing organisms to the
most used effect endpoints in the laboratory contaminants. The bioaccumulation tests were
bioassays. The acute tests (15 min to 96 h period also used for quantified the metal accumulation
of test) were frequently used to obtain a rapid (bio concentration / bioaccumulation factor
dose – response (L(E)C50). The long term (BCF) factors) in the fish or plants tissues in
effects were underlined through chronic fish test controlled laboratory conditions or directly in
(30-90 days) or ciliates tests (24h). The acute surface water / sediment.
effects are the most reproducible and can be In addition, the enzymatic activities and /or
calculated with high reliability. Static or semi- histology modifications were assessed to
static systems were performed for the acute evaluate the effects of toxicants on fish.
Microbacterium
sp.,Brevundimonas diminuta,
Citrobacter freundii,
Comamonas testosterroni,
MARA test (Microbial
Enterococcus casseliflavus, Chemical /
Array for toxicity Risk acute Microbial growth inhibition, MTC 18 h, 30°C
Delftia acidovorans, Kurthia water samples
Assessment)
gibsonii, Sthaphilococcus
warnerii, Pseudomonas
aurantiaca,
Serratia rubidae, Pichia
anomalia
Geno toxicity test Chemical / Effects on genes expression
Escherichia coli acute 18h, 37°C
SOS Chromotest water samples (β-galactosidase)
Terrestrial plants
Lepidium sativum, Chemical, soil,
SR EN ISO 11269, Germination and root growth
Sinapis alba, Sorghum sludge, waste, acute 72 h, 25°C
Phytotoxkit inhibition, EC50
saccharatum sediment, water
Note: LC50 / EC50 / IC50 – median lethal / effect / inhibitory concentration; MATC - Maximum Acceptable Toxicant Concentration in aquatic
systems, MTC – microbial toxic concentration
Our data accumulated over the time, allowed us concentration - UTC) [28]. The smallest EC50
to conclude that the use of microbiotests (with between algae and crustacean will be used for
algae, crustacean and bacteria) have had a good acute LC50 biotests with fish.
reproducibility, relevant and specific results. If no mortality was observed, the test was
Otherwise, because the vertebrate represents the stopped and LC50 greater than (>) the UTC
final target of aquatic pollution, the toxicity tests value was reported. If the mortality was
with fish have an important role in risk observed the test continue at low concentrations
assessment, therefore this can be used in using a dilution factor until no mortality was
combination with microbiotests. observed.
The multi - species tests can offer relevant data
concerning the chemicals risk that may lead to
an economical and efficient assessment of acute
and chronic toxicity [13, 16]. In the Fig. 3 the
acute testing plan of chemicals under REACH
Regulation applied in the ecotoxicity studies was
presented.
Firstly, the data about the chemical / chemical
product/ environment sample were collected to
have a predicted vision about their toxicity in
water. Considering the Klimisch score to
evaluate the quality of experimental data [27]
the need for future information and what test
should be performed were established. If the
data are not sufficient for risk assessment, new
tests are needed.
Secondly, tests using algae, Daphnia and
bacteria were performed to obtain de lowest
EC50 (effective concentration with inhibition /
mortality/ immobilization effect for 50% of Fig. 3. Acute toxicity laboratory testing plan
tested organisms set as upper threshold under REACH [13]
The results led to predict the maximum allowable environmental risk coefficients [10, 11].
surface water concentrations, predicted non effect By regularly consulting of the stage of
concentrations and also to estimate the knowledge’s in the research field, our
Romanian Journal of Ecology & Environmental Chemistry ● No.1 ● 2019
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31
S. Gheorghe et. al.: Anthropogenic pollutants: 10 years of progress in ecotoxicological studies and aquatic risk assessment
ecotoxicity topics are fitted in the international and algae methods are accredited and required
development requirements. A statistical analysis by the economic media, their frequency in the
of the frequency uses of fish in toxicity tests last period was higher compared to the other
during the last 10 years revealed a 60% decrease species (such as bacteria and other small
in 2018 compared to 2009 due to the organisms) (Figure 5). Also, ecotoxicity and risk
implementation of new testing strategies and the assessment studies have included test batteries
use of invertebrates in the first stage of toxicity that specifically targeted these three species,
assessment (Figure 4). Because the fish, daphnia their use being internationally regulated.
Fig. 4. The fish toxicity test trend in the period Fig. 5. Frequency of biotests use in period 2009-
2009-2018 2018
In this context the next paper sections will The ecotoxicity studies with single species tests
present the most important results obtained in were performed in case of: i) chronic tests with
the least 10 years of ecotoxicological research. fish (Cyprinus carpio) to highlight the chemical
The studies were focused on ecotoxicological bioconcentration or sublethal effects using
characterization of chemicals or environmental enzymatic activity or protein expression; ii)
samples using bio-indicators or biomarkers. efficiency tests with algae (Selenastrum
Generally, the aquatic risk assessment capricornutum / Pseudokirchneriella subcapita)
representing de the end point of ecotoxicological developed for the algaecide efficiency test; ii) or
studies. in research studies when the species are selected
to be bio-indicator of natural ecosystem.
RESULTS AND DISCUSSIONS To assess the toxic hazard of contaminated
Aquatic toxicity - Mono-species experiments sediment samples, meiobenthic ostracods
The single species toxicity tests were Heterocypris incongruens were used as bio-
appropriate for analyzing toxicological effects indicator for a six days’ chronic test.
on individual characteristics of species such as The percentage of organism’s growth-inhibition
mortality or growth, but with limited was assessed. The research study involved the
significance for the consequences on the natural spatial and temporal monitoring of some
ecosystem. physical, chemical and biological indicators both
Before 2008, our laboratory performed LC50 from water and sediment and subsequently a
individual species tests using fresh water bio correlation with the laboratory ostracods toxicity
indicators such as Cyprinus carpio and Daphnia test was made. The toxicity tests presented a
magna to evaluate the acute or chronic toxicities realistic view of field effects. The effects on
of various chemicals (pesticides, metals or other benthic organisms were directly influenced by a
inorganic or organic compounds) [11, 29]. mixed content of contaminants from sediment,
Currently, this tests are frequently request by especially in those Danube areas where the
producers or control authorities for chemical anthropogenic activities were more intense
products authorization (such as biocide, (Ivancea and Murighiol). This fact was
industrial products or industrial wastes) or correlated with the benthic diversity from this
environmental samples toxicity control. area [6]. Others specific studies were performed
to evaluate the subacute effects of chemicals In the context of new interest of the European
using the biomarkers expressed in living Commission concerning the improvement of the
organism such as: antioxidant defense enzymes, environmental performance of products and the
hepatic enzymes or protein expression. increase the demand for more sustainable goods
Chemicals such as: benzenthonium chloride and production technologies by 2030, our
(quaternary ammonium compound - cationic laboratory developed ecotoxicity test suitable for
surfactant), cocamidopropyl betaine each class of chemicals including the biocide.
(zwitterionic compound amphoteric), atrazine The use of two species from the same trophic
and mevinphos (organophosphorus pesticides), level test is exceptionally applied to showed the
trichlorfon and monolinurone (organochlorine sensibility difference. A study of biocides lethal
pesticides) were tested. The level of oxidative and sub lethal effects was performed using two
stress biomarkers (superoxide dismutase – SOD, representative crustacean Daphnia magna
catalase - CAT, glutathione peroxidase - GPx, (pelagic system) and Heterocypris incongruens
glutathione reductase - GR, glutathione S (benthic system). The tests showed a significant
transferase - GST, glucose -6- phosphate difference of sensitivity between species. The
dehidrogenase – G6PDH) and of lipid biocides induced a very toxic or toxic effect on
peroxidation - LPO were evaluated. After 96h of Daphnia magna. On the contrary, the observed
exposure, the toxicity of surfactants (1 mg/l) is ostracods lethal effects were less than 50%
selective at organs levels. The liver and gills mortality and no significant growth inhibitions
were the most affected by toxic action. The were observed [35].
enzymatic biomarkers assessment highlighted Aquatic toxicity - Three-species experiments
deficit in reactive oxygen species balance The phytotoxic effects of some contaminated
followed by oxidative stress in fish organs [30, environmental matrices such as metal /
31]. Also, it was proved that acute intoxication petroleum products polluted soil or sewage
of fish with atrazine (10 mg/l) and mevinphos sludge were test for three days in the same
(0.1 mg/l) inhibit the enzymatic activity of condition using three higher plants Sorghum
MnSOD and CuZnSOD, the detoxifying saccharatum, Lepidium sativum and Sinapis
function of SOD enzyme being discredited, alba. The inhibition of germination and roots
resulting in high death rate during the test [32]. growth inhibition were monitored. The
Also, trichlorphon (1 mg/l) and monolinuron anthropogenic pollutants from the contaminated
(1.5 mg/l) could induce an easy change in samples determined different sensitivity
antioxidant enzymatic activity in fish liver [33]. responses. The tests could be use as routine
The hepatic enzymes activity of glutamic toxicity screening tests to provide rapid
oxaloacetic transaminase (GOT) and glutamic informations for soil contamination [36], waste
pyruvic transaminase (GPT) were used as water or sludge reuse [37, 38], bioremediation
biomarkers of acute toxicity of some industrial and environmental risk assessment.
products (fluids used in metalic processing). The Aquatic toxicity - Multi-species experiments
tests suggested that the products do not have The biotests battery is based on acute sensitivity
significant effect on detoxification system and responses of a various representative species of
the mixtures could be considerate ecological trophic chain in order to predict the natural
products [34]. system behavior to the chemical contamination
The protein expression in algae extracts were distress. The multi-species tests include species
used to evaluate the toxicity of two pesticides of algae, crustacean, fish, and also rotifers,
(bentazon and 4-chloraniline). plants or bacteria. Thereby, a series of specific
After seven days of test, a differentiated laboratory studies were performed to assess the
expression of cellular protein was showed toxicological effects of cationic and amphoteric
according to chemical structure of contaminant, surfactants [39], pharmaceutical compounds
concentration and exposure time. New (analgesics, antibiotics, nervous stimulators)
adaptation or defense proteins were identified in [16], pesticides [11] and other organic
the protein profile comparative with the control. compounds complexed with metals [40]. The
The results are in the publication process. final results allowed us: i) to reveled a
Aquatic toxicity - Two-species experiments differentiated response for each trophic chain
More results and final conclusion of metals investigate the accumulation of heavy metals.
bioconcentration studies are in publication The study shows that the Lemna minor L.,
process. Other studies used lichens Parrmelia accumulates considerable amounts of Zn, Cu
spp. as biosensors in the monitoring of and Ni without any symptoms of toxicity [44].
environmental pollution with heavy metals (Cd, Risk assessment studies
Zn, Pb). The lichens have classified the polluted The ecosystem health is usually based on risk
sites by the characteristics of bioaccumulation of assessment considering the chemical
metals in lichens. Based on the corticols lichens concentration in the environment (PEC) and the
specificity a correlation index of air quality in impact on organisms (PNEC). A ratio of
the interest geographical areas was calculated. PEC/PNEC < 1 indicated no aquatic risk and no
Also, the capacity to accumulate metal in the future assessments is deemed necessary.
lichen communities has been used to identify the Our studies on chemical risk assessment applied
level of air pollution due road traffic. Based on the international methodology adjusted to our
the degree of heavy metal accumulation such as laboratory conditions and indigenous organisms.
Cd, Cr, Cu, Fe, Mn, Pb, Ni and Zn in the After 2009, the final results of our research
Parmelia spp. lichens, a correlation has been studies was to obtain an risk study. The
made, with the road traffic. According to the integrated chemicals risk studies involved data
metal bioaccumulation degree in lichens tissue, of chemical detection in laboratory and field
it has been established that the sources such as condition, toxicity experimental data of
traffic from the outskirts of cities, from the chemical on living organisms, biodegradability,
perimeter of gas stations and of county roads environmental contaminated samples collection
continuously spread products which contain (toxicity testing and chemical detection), sub
these elements, into the atmosphere [43]. lethal effects evaluation, selection of the most
The aquatic plants such as Lemna minor L. has sensitive organisms and risk level establishment
been reported to accumulate toxic metals and is (Fig. 6).
therefore used as an experimental model to
REFERENCES
1. LYUBENOVA, M., BOTEVA, S., Biotests in Ecotoxicology: Current Practice and
Problems, chapter 7 in „Toxicology - New Aspects to This Scientific Conundrum” edited by
Marcelo Larramendy and Sonia Soloneski, ISBN: 978-953-51-2717-8, IntechOpen, 2016,
p.147.
2. GHEORGHE, S., LUCACIU, I., PASCU, L., J. Environ. Prot. Ecol., 13, no.1, 2012, p. 155.
3. GHEORGHE, S., LUCACIU, I., GRUMAZ, R., J. Environ. Prot. Ecol., 12, no. 3A, 2011, p.
1525.
4. LILLICRAP, A., BELANGER, S., BURDEN, N., DAVID, D.P., EMBRY, M.R., et al.,
Environ. Toxicol. Chem., 35, 2016, p. 2637.
5. SZKLAREK, S, STOLARSKA, M., WAGNER, I., MANKIEWICZ-BOCZEK, J., Environ.
Monit. Assess., 187, no. 16, 2015, p. 15.
6. STOICA, C., GHEORGHE, S., LUCACIU, EI., STANESCU, E., PAUN, CI.,
NICULESCU, LD., Soil Sediment. Contam., 23, no.7, 2014, p. 763.
7. PERSOONE, G., WADHIA, K., Comparison Between Toxkit Microbiotests and Standard
Tests, DOI: 10.1007/978-0-387-88959-7_23, In book: Ecotoxicological Characterization of
Waste, 2009.
8. FENT, K., WESTON, A.A., CAMINADA, D., Aquatic Toxicology, 76, 2006, p. 122.
9. STRAUB, J. O., Toxicology Letter, 131, 2001, p. 137.
10. EMEA, Guideline on the environmental risk assessment of medicinal products for human
use; The European Agency for the Evaluation of Medicinal Products, London, England,
EMEA/CHMP/SWP/4447/00, http://www.emea.eu.int/pdfs/human/swp/444700en.pdf.
11. GHEORGHE, S., LUCACIU, I., PAUN, I., STOICA, C., STANESCU, E., J. Environ. Prot.
Ecol., 15, no. 3, 2014, p. 878.
12. GHEORGHE S., LUCACIU, I., GRUMAZ, R., SGEM 2010, 21-26.06.2010, Albena,
Bulgaria., p. 669.
13. GHEORGHE, S., LUCACIU, I., STANESCU, E., STOICA, C., J. Environ. Prot. Ecol., 14,
no.2, 2013, p. 601.
14. LUCACIU, I., LUCA, M., ORBAN, T., GHEORGHE, S., GRUMAZ, R., Interntional
Symposium – The Environment and Industry, Bucharest, SIMI 28-30.09.2009, p. 214.
15. GHEORGHE, S., STOICA, C., PAUN, I., LUCACIU, I., NITA-LAZAR, M., CRISTOFOR,
S., J. Environ. Prot. Ecol., 17, no.1, 2016, p. 171.
16. GHEORGHE, S., PETRE, J., LUCACIU, I., STOICA, C., NITA-LAZAR, M., Environ.
Monit. Assess., 188, no. 6, 2016, p. 379.
17. VOSNIAKOS, F., PETRE, J., PASCU, L., VASILE, G., IANCU, V., STANILOAE, D.
NICOLAU, M., CRUCERU, L., GOLUMBEANU, M., Fresenius Environmental Bulletin,
19, no. 1, 2010, p. 21-31.
18. IANCU, V.I., GALAON, T., PETRE, J., CRUCERU, L., PASCU, L., 15th International
Multidisciplinary Scientific GeoConference SGEM 2015.
19. IANCU, V.I., PETRE, J., CRUCERU, L., Environ. Prot. Ecol., 12, no. 3, 2011, p. 833.
20. IANCU, V.I., GALAON, T., PETRE, J., CRUCERU, L., PASCU, L. FLORENTINA, L.,
CAROL B., Rev. Chim. (Bucharest), 67, no. 8, 2014, p. 1484.
21. GALAON, T., PETRE, J., IANCU, V.I., CRUCERU, L., VASILE, G., PASCU, L.F.,
BLAZIU L.C., Rev. Chim. (Bucharest), 67, no.8, 2016, p.1474.
22. PETRE, J., IANCU, V., VASILE, G., ALBU, F., NICULESCU, M., NICULAE, A.
CRUCERU, L., NICOLAU, M., The GeoConference on Ecology, Economics, Education
and Legislation, vol. 1, 2013, p. 71.
23. VASILE, GG., GHEORGHE, S., ENE, C., SERBAN, EA., STOICA, C., Rev. Chim.
(Bucharest), 70, no.1, 2019, p. 263.
24. VASILE, G., TANASE, IG., Rev. Roum. Chim., 53, no. 11, 2008, p.1041.
25. DINU, C.,VASILE, G.,CRUCERU, L., J. Environ. Prot. Ecol., 14, no. 4, 2013, p. 1515.
26. GHEORGHE, S., LUCACIU, I., PAUN, I., STOICA, C., STANESCU, E., book chapter 4,
Biodegradation – Life Science, edited by Dr. Rolando Chamy and Francisca Rosenkranz,
ISBN 978-953-51-1154-2, Editor InTech DTP, 2013, p. 83.
27. KLIMISCH, H.J., ANDREAE, M., TILLMANN, U., Regulatory Toxicology and
Pharmacology, 25, 1997, p.1.
28. ESAC, (2005). Peer Review of European Commission, Proposal for a new testing strategy
(Step-down approach) to reduce the use of fish in acute aquatic toxicity testing, Joint
Research Centre.
29. GHEORGHE, S., STOICA, C., NITA-LAZAR, M., GEANINA, VASILE, G., LUCACIU,
I., Metals toxic effects in aquatic ecosystems: modulators of water quality, in Water Quality,
Book Chapter 4, In: Water Quality, InTech, Hlanganani Tutu(Ed.), DOI: 10.5772/65744,
2017, p. 59.
30. GHEORGHE, S., LUCACIU, I., DINISCHIOTU, A., CATALINA, S., IULIANA, P.,
Interntional Symposium, The Environment and Industry, Bucharest, SIMI 16-18.10.2011,
vol. 2, p. 232.
31. IVAN (GHEORGHE), S., STAN, M., DAMACHE, G., STOICA, C., STĂNESCU, E.,
IRINA, L., DINISCHIOTU, A., Effects of Cocamidopropyl betaine on Oxidative stress
biomarkers of the Cyprinus carpio sp., 3rd SETAC CEE Annual Meeting, 17-19 September
2012, Cracovia, Polonia, “Ecotoxicology revisited”, p. 33.
32. IVAN, S., LUCACIU, I., RUSU, G., VASILE, I., J. Environ. Prot. Ecol., 11, no. 1, 2010, p.
247.
33. GHEORGHE, S., RUSU, G., LUCACIU, I., IANCU, V., J. Environ. Prot. Ecol., 12, no. 3A,
2011, p. 1301.
34. LUCACIU, I., GHEORGHE, S., PETRE, J., GRUMAZ, R., Interntional Symposium – The
Environment and Industry, Bucharest, SIMI 28-30.09. 2009, p. 321.
35. GHEORGHE, S., STOICA, C., LUCACIU, I., BANCIU, A., NITA-LAZAR, M., Rev.
Chim. (Bucharest), 70, no.1, 2019, p. 307.
36. GHEORGHE, S., VASILE, G., IANCU, V., LUCACIU, I. Romanian Journal of Plant
Protection 4, 2013, p. 43.
37. GHEORGHE, S., PAUN, I., BANCIU, A., VASILE, G., IANCU, V. I., NITA-LAZAR, M.,
Interntional Symposium - The Environment and Industry, Bucharest, SIMI 29-30.10.2015.
38. GHEORGHE, S., VASILE, GG., STOICA, C., NITA-LAZAR, M., LUCACIU, I.,
BANCIU, A., Rev. Chim. (Bucharest), 67, no. 8, 2016, p. 1469.
39. GHEORGHE, S., LUCACIU, I., GRUMAZ, R., STOICA, C., J. Environ. Prot. Ecol., 13,
no. 2, 2012, p. 541.
40. CRAIOVEANU, MG., GHEORGHE, S., LUCACIU, I., STOICA, L., CONSTANTIN, C.,
Rev. Chim. (Bucharest), 65, no.3, 2014, p. 339.
41. PERSOONE, G., et al., Environ. Toxicol., 18, no. 6, 2003, p. 395.
42. GHEORGHE, S., VASILE, GG., GLIGOR, C., LUCACIU, IE., NITA-LAZAR, M., Rev.
Chim. (Bucharest), 68, no. 8, 2017, p. 1711.
43. MÂŞU, S., Animal Sci and Biotechn, 50, no.1, 2017.
44. NEIDONI, D.-G., NICORESCU, V., ANDRES, L., IHOS, M., LEHR, C.B., Accumulation
of toxic metals in aquatic plants, DOI: http://doi.org/10.21698/simi.2018.ab30.
Advanced Oxidation Processes (AOPs) alternative methods for degradation of toxic pollutants
from wastewater
National Research and Development Institute for Industrial Ecology ECOIND Bucharest, 71-73 Drumul Podul Dambovitei, 060652,
Bucharest, Romania
*Corresponding author (e-mail): [email protected]
The degradation of some toxic pollutants like chlorobenzenes, nitrobenzene and 4-chloroaniline in
various advanced oxidation system such as: UV/ H2O2, UV-VIS/Fe-TiO2 and UV-VIS/TiO2/ H2O2
were studied. The influence of working conditions (pH0, H2O2 dose, photocatalyst dose, pollutant
initial concentration and irradiation time) on pollutant degradation rate constant and efficiency
were investigated. For any studied advanced oxidation systems, pollutant degradation followed a
pseudo first order kinetics. The degradation pathway of chlorinated and nitro aromatic pollutants
includes initial •OH radicals attack to aromatic ring with hydroxylated intermediates formation,
followed by their step by step oxidation up to carboxylic acids. Inorganic ions like Cl-, NO3-, NH4+
are also formed as mineralization products.
Water is essential for life and although Advanced Oxidation Processes (AOPs) based on
approximately 70% of the terrestrial area is photochemical and photocatalytic approaches
covered with water, only a small fraction (2.5%) seem to be very promising as viable alternatives
is freshwater compatible with terrestrial life [1]. for appliance on wastewater treatments of
This low stock of available freshwater is being different sources.
accompanied by an increase of human These approaches allow the transformation of
consumption as consequence of demographic pollutants into less toxic substances and/or with
growth, industry demand and improved living structural features that are more biodegradable
conditions. It is even estimated that in some [7] and are based on the combination of
world regions, the water use has risen twice oxidizing agents with an appropriated catalyst
faster than the human population. In fact, half of and/or light [7, 8].
the European countries are already facing water These processes may be of particular interest for
stress and it is estimated that within the next 50 the effluent’s treatment containing highly toxic
years more than 40% of the population compounds like emerging pollutants, and for
worldwide will face water stress or scarcity [1- which the biological processes might not be
3]. All this together represents a solemn appropriate [7].
inducement to achieve sustainable management AOPs involves specific chemical reactions that
options of the water resources [1]. The human generate highly reactive chemical oxidizing
being plays a major role on the issue of species which are capable to oxidize and
pollutants released in wastewater, which mineralize almost any organic molecule, even
includes heavy metals, antibiotics, pesticides, the most recalcitrant molecules to CO2 and
polyaromatic hydrocarbons, halogenated inorganic ions [7–10]. These processes generally
compounds, and endocrine disruptors, most of involve use of light. The degradation by
them known as emerging pollutants [4, 5]. photochemical reactions of organic pollutants
As conventional methods applied in wastewater present in water media can be achieved, but the
treatment are limited and their associated degradation degree is dependent on their
operational and capital costs are high, the issue chemical structure [11]. The AOPs efficiency
of emerging pollutants removal from tends to be maximized when the use of an
environment is not completely solved [6]. appropriate catalyst is considered. Although
The results achieved in latest years showed that there are different oxidative systems, the
Romanian Journal of Ecology & Environmental Chemistry ● No.1 ● 2019
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39
I. Nitoi et. al.: Advanced Oxidation Processes (AOPs) alternative methods for degradation of toxic pollutants from wastewater
production of powerful oxidizing agents, such as certain types of wastewater and operational
hydroxyl radical (HO•), is the main objective of conditions. The AOPs which will be reviewed in
most AOPs, since these oxidizing agents react the following subsections are hydrogen peroxide
very fast [9]. Also, their nonselective attack to assisted photocatalysis (UV/H2O2) and
organic compounds is a useful attribute for photocatalytic process in the following systems:
wastewater treatment and a potential solution of UV/0.5Fe-TiO2 and UV/TiO2/H2O2. Target
pollution issues. These oxidizing species can act studied pollutants have been halogenated
at various levels such as hydrogen abstraction aromatic compounds like chlorobenzenes (CB)
from aliphatic carbon, electron transfer or by and 4-chloraniline(4CLA) and nitro aromatic
addition to double bonds and aromatic rings derivative, respectively nitrobenzene(NB), all of
[7,10,12]. This type of reactions generates them included in the emerging compounds lists.
organic radicals as intermediates, which then
undergo additional reactions, ending in final EXPERIMENTAL PART
products [13]. The hydroxyl radicals can react The pollutants photodegradation was carried out
with a vast range of organic compounds at rates in a Laboratory-UV-Reactor system (Heraeus)
often nearby the diffusion-controlled limit with medium-pressure mercury lamps which
[7,14,15]. emit in the range of 200-280 nm (UVC) (TQ150
The versatility of the AOPs is likewise enhanced lamp), and respectively in the range 300-500nm
by the fact that exist different ways to produce (UV-VIS) (TQ150-Z2).
hydroxyl radicals, facilitating the compliance
with specific treatment requirements in The lamp equipped with a quartz cooling jacket
accordance with effluent’s characteristics [9,10]. was immersed in the center of the reactor
The AOPs as a photochemical process can occur containing the pollutant solution. The photon’s
according two main ways: homogeneous (when flow of the emitted radiations was determined
both reactants and pollutants are in the same by ferrioxalate actinometry and values I0 = (5.3-
phase) and heterogeneous systems (when 6) x10-6 einstein s-1 were found.
reactants and catalysts are in different phases. In Solutions with CB concentrations between
general, photocatalytic process can be conducted 0.1x10-3 - 1.8x10-3M and of H2O2 between
under visible-light irradiation either with or 0.3x10-2 – 9x10-2 M, which correspond to molar
without the presence of UV radiation [7]. The ratio [H2O2]/[CB] in the range 5/1-50/1, were
region of the electromagnetic spectrum used to used in peroxide assisted photolysis
the photochemical process is dependent on the experiments. In the case of photocalysis
reactant/catalyst absorption features [10]. experiments in UV-VIS/metal doped TiO2,
Methods based on ultraviolet light (UV), and solutions with (0.37-8.45) x 10-4 M NB content
H2O2 use the photolysis of this oxidizing agent were photo-oxidized in the following working
to produce hydroxyl radicals [9,10]. conditions: pH= 4-10; photo catalyst dose = 50-
Additionally, other applied methods like 500 mg/L; irradiation time = 30 – 240 minutes.
heterogeneous photocatalysis, are based on the Experiments performed in UV/TiO2/H2O2system
use of a wide-band-gap metal oxide were conducted with various concentration of
semiconductor without or with addition of H2O2 H2O2 between 1 x 10-4 – 1 x 10-2 M, at fixed
to retardation of electrons-holes recombination pH=4, TiO2 dose 150 mg/L and 1.42 x 10-3 M 4-
process which assures enhance of catalyst CLA concentration. Prior to irradiation, the
photoactivity, respectively, and irradiation with required powder photocatalyst dose was added
UV–VIS light. Both processes are of interest due to samples, and the solutions were bubbled with
to fact that the homogenous system has air (50L/hour) for 30 min in the dark for both
advantages of hydrogen peroxide which is easy photocatalytic systems studied. The sample’s pH
to handle and environmentally safe, since in the was controlled using a Consort C532 pH-meter.
heterogeneous system sunlight can be used, The solution within the reactor was
especially for doped semiconductors, which homogenized continuously using a magnetic
brings a set of economic benefits. circulation pump.
There are several types of AOPs used in The concentration of CB and NB were
WWTPs, each one of them can be effective in determined via Gas Chromatography couple
Kinetics of CB degradation in UV/ H2O2 system Using the slopes of these straight-lines the rate
and H2O2 concentration influence constants kCB and kCl- were calculated. In Figure
The photodegradation kinetic of CB in the 2 there is presented the evolution of the rate
presence of H2O2, at pH=7, under UV irradiation constants versus the hydrogen peroxide
verified a pseudo-first-order reaction rate with concentration.
respect to the pollutant and chloride ion, one of It is observed that values of kCl-are always lower
the mineralization final product [28]. Figure 1 than values of kCB, suggesting that the Cl-
presents the kinetics monitored at pH=7, for a liberation does not take place at the same time
sample with an initial CB concentration of with the attack of •OH radicals on CB
1.8x10-3M at various H2O2 concentrations molecules. In a first stage chlorinated organic
corresponding to a molar ratio of [H2O2]/[CB] intermediates are formed, which are further
between 0/1-50/1. releasing chlorine as Cl- ion. In the range of low
H2O2 concentration the both rate constants
Fig. 1. The pseudo-first-order kinetics at various increase linearly with H2O2concentration. On the
H2O2 concentration of contrary, at high H2O2 concentration, these rate
a) CB degradation and b) Cl- formation constants decrease due to the fact that •OH radi-
cals are scavenged by the excess of H2O2.
0 Between these two ranges kCB and kCl- reach
their maximum values 15x10-4 s-1 and 9x10-4 s-1,
[H2O2]/[CB]
-1 respectively, for a short interval of H2O2
0/1 concentration (5.4x10-2 - 7.2x10-2 M)
-2
0/1 R2 = 0.9808
corresponding to a [H2O2]/[CB] molar ratio 30/1 –
5/1 R2 = 0.9768
40/1. The extrapolation of the linear part leads to
ln[CB t]/[CB 0]
5/1
10/1 R2 = 0.9982
-3 20/1 R2 = 0.9798 10/1 rate constants values near to those experimentally
25/1 R2 = 0.9982
50/1 obtained in the absence of photosensitizer,
30/1 R2 = 0.9871
-4 via CB direct photolysis.
40/1 R2 = 0.9911 20/1
50/1 R2 = 0.9592
Therefore, this behavior can be described by the
25/1
-5 40/1 kinetic equation: kCB = k0 + k∙OH [H2O2], in
a)
30/1
which kCB is the rate constant of CB degradation
-6 determined for various initial H2O2
0 300 600 900 1200 1500 1800 2100 2400 2700 3000 3300 3600
Time, s
concentrations, k0is the rate constant of CB
photolysis and k∙OH the second-order rate
constant of CB oxidation by OH radicals. In Fig.
3.5 [H2O2]/[CB]
30/1
2 the first linear part represents in fact the plot of
0/1 R2 = 0.9811
2
5/1 R = 0.9989
40/1 before mentioned kinetic equation. A similar
3
b)
10/1 R2 = 0.9955 25/1
equation is valid also for Cl- formation. Some
2.5
2
20/1 R = 0.9981 20/1 other researchers [24] using similar molar ratio
2
25/1 R = 0.9975 [H2O2]/[CB] than in our experiments found that
-ln(1-Cl t]/[Cl f])
10/1
1.5 50/1 R2 = 0.99 mediated degradation takes place in their
irradiated system. However, a similar behavior
1 5/1
of the system to that we reported was presented
0/1 by the same authors in their earlier work, but it
0.5
was not explained. We consider that the
0
contribution of direct photolysis to CB degrada-
0 300 600 900 1200 1500 1800 2100 2400 2700 3000 3300 3600 tion is due to the manipulation of the samples at
Time, s the room light before the irradiation in the photo
reactor.
Fig.2. The influence of H2O2 concentration on the rate constants kCB and kCl-.
Reaction mechanism attack of •OH radicals on CB, but in a further
Many scientists studied the photodegradation of stage, oxidizing the chlorophenols to
CB in various experimental conditions, like hydroquinone and three hydroxibenzene.
photolysis, UV/H2O2, UV/O3, UV/TiO2 but only According to our studies, formed intermediates
some of them analyzed the systems in order to compete with CB for •OH radicals consumption
obtain information on the reaction mechanism as experimental results are emphasizing by the
[24,27,29]. We tried to correlate these data with rate constant values. The rate constants values
our results to obtain a general plausible reaction decrease with an increase in the initial CB
mechanism. This includesH2O2photolysis under concentration, at constant [H2O2]/[CB] molar
UVC irradiation and generation of •OH radicals ratio. The before cited authors [27] reported that
which by addition to the double bound, or by hydroquinone is oxidized to benzoquinone
hydrogen abstraction attacks the CB leading to which is cleaved in muconic acid and then under
various intermediates [30], the majority being •OH radical attack leads to maleic, fumaric and
hydroxylated aromatic compounds. Using GS- oxalic acids. In the favor of this presumption we
MS analysis, some authors identified have a qualitative argument: by the appearance
chlorophenol, chlorobiphenyl and of a reddish-brown color, characteristic to
dichlorobiphenyl [24]. Other authors studding benzoquinone, which under prolonged exposure
the CB degradation in UV/TiO2 systemrevealed time bleaches due to carboxylic acids formation.
that o- and p-chlorophenol are further oxidized to The presence of many stable intermediates is
hydroquinone and catechol [29]. Using flash justified by the low removal yield of total
photolysis/HPLC technique for the degradation organic carbon (TOC) of about 14%, determined
of chlorophenol in the presence of H2O2, have after irradiation, in our experiments [28].
been put in to evidence the same hydroxylated
compounds [27]. It has also been shown that Photocatalytic pollutant degradation
increase of H2O2 concentration changes the These processes use a semiconductor metal
product distribution, catechol concentration drop oxide as catalyst and oxygen as oxidizing agent
in the favor of hydroquinone and 1,2,4-three- [31]. Many catalysts have been so far tested,
hydroxybenzene. The formation of these although only TiO2 in the anatase form seems to
unchlorinated intermediates agrees with our havethe most interesting attributes such as high
results which show the total liberation of the stability, good performance and low cost [32,
organic bound chlorine as HCl, during 33]. The initiating step in the photocatalytic
irradiation. On the other hand, our finding that process is the absorption of the radiation with
kCB>kCl- demonstrates that Cl- ions liberation the formation of electron-hole pairs:
does not take place simultaneously with the TiO2+hν→e−+h+ (4)
The considerable reducing power of formed other oxidizing agents. There is another
electrons allows them to reduce some metals and advantage related to the use of this type of
dissolved oxygen with the formation of the oxidant when solar energy is the photon source.
superoxide radical ion O2•− whereas remaining This is related to increase of the photocatalytic
holes are capable of oxidizing adsorbed H2O or reaction rate which assures proportionally
OH − to reactive •OH radicals: decrease of photo reactor dimensions and
TiO2(h+) + H2Oad→TiO2+•OHad+H+ (5) dramatically decrease of overall costs.
TiO2(h+) + OH−ad →TiO2+•OHad (6) Hydrogen peroxide is the obvious candidate. It
These reactions are of great importance in can increase the efficiency of the process at high
oxidative degradation processes due to the high irradiance and it has been tested with a large
concentration of H2O and OH− adsorbed on the number of compounds. Also, it is a very
catalyst particle surface. commonly used chemical and, therefore, very
cheap. Being an electron acceptor, hydrogen
Some adsorbed substrate can be directly peroxide reacts with conduction band electrons
oxidized by electron transfer: to generate hydroxyl radicals, which are
TiO2(h+) +RXad→TiO2+RXad•+ (7) required for the photo mineralization of organic
Unfortunately, a significant part of electron-hole pollutants.
pairs recombine thus reducing the quantum yield
of photocatalytic process. In the most part of the H2O2+e-→•OH+HO- (8)
works devoted to the photocatalysis the possible The following reactions (eqs. 9,10) can also
exploitation of the wavelengths of the solar produce •OH (eq.10, which requires λ < 300 nm,
spectrum is stressed. However, this is only does not take place with solar radiation).
partially true since the overlapping between the
absorption spectrum of TiO2 and that of the sun H2O2 +O2•- → •OH + HO- + O2 (9)
at ground is rather poor [33]. Intensive H2O2+hν → 2 •OH (10)
researches are carried out worldwide to obtain The effect of this electron acceptor deserves
modified (doped) TiO2 with broader absorption some further comment. In some cases, the
spectrum and characterized by higher quantum addition was found to be beneficial, increasing
yield. the degradation rate. The effect depends on the
Another strategy for improving the H2O2 concentration, generally showing an
photocatalytic activity consists of inhibiting e- optimum range of concentration. At higher
/h+ recombination by addition of other concentrations the improvement starts to
(irreversible) electron acceptors to the reaction. decrease. Whereas this beneficial effect can
Outstanding enhancement of the rate of easily be explained in terms of prevention of
degradation of various organic contaminants electron/hole recombination and additional •OH
through the use of inorganic peroxides has been production through reactions 8,9 and 10,
demonstrated [34,35]. Beside increasing of the inhibition could be explained in terms of TiO2
number of trapped e- in the e-/h+ pairs and, surface modification by H2O2 adsorption,
consequently, avoid their recombination, scavenging of photo produced holes (eq.11) and
peroxides addition could have also several reaction with hydroxyl radicals (eq.12).
different effects: • generation of more •OH and
other oxidizing species; • increase the oxidation H2O2+ 2h+ → O2+ 2H+ (11)
rate of intermediate compounds; • avoiding H2O2+•OH →H2O+ HO2•- (12)
problems caused by a low O2 concentration. The inhibition of adsorption not only depends on
It must be mentioned here that in highly toxic the characteristics of the pollutant, but also on
wastewater where degradation of organic the H2O2/pollutant concentration ratio.
pollutants is the major concern, the addition of
an inorganic anion to enhance the organic Metal doped TiO2 assisted photocatalytic
degradation rate may often be justified. degradation of NB under UV-VIS irradiation
For better results, these additives should fulfill Because of the high energy band gap (3.2 eV),
the following criteria: dissociate into harmless titania can be activated only by ultraviolet (UV)
by-products and lead to the formation of •OH or light [36]. In addition, low photo quantum
efficiency and high recombination of electron- titania matrix which acts as electron trap, and
hole pairs restrict the titania application. In order also assures enhancement of photocatalytic
to improve optical absorption and photocatalytic activity [41]. Since, charges recombination rate
activity of TiO2 many attempts have been made depends on dopant type and its concentration,
[37]. One feasible approach consists of doping the photocatalytic activity of undoped and Fe,
oxide semiconductor with metal [38-40]. Co, Ni doped TiO2 in degradation of NB from
The degradation of NB from aqueous solution aqueous solution (2.52x10-4 M) was investigated
under UV-VIS irradiation using heavy metal [42]. The dopant concentrations, reported to
doped titania was investigated. Dopant type (Fe, TiO2 were 0.5; 1; 2; 5wt.%. All experiments
Ni, Co) and their concentration were assessed in were performed at pH=7, with 100 mg/L
relation with pollutant degradation kinetic. Also catalyst dose and irradiation time between 30-
catalyst dose, pH and initial NB concentration 240 min. The efficiency of pollutant degradation
influence on degradation efficiency were was assessed by determination of NB
evaluated in order to set up the optimal working concentration vs. irradiation time in treated
conditions which assure pollutant’s advanced samples. Kinetic curves obtained in various
degradation. working conditions, were linearized by a first
Photocatalytic activity of metal doped TiO2 order reaction kinetic. From the slope of each
The trapping of electrons can inhibit equation pseudo-first order rate constants, kNB
electron/hole recombination during irradiation, were calculated. Pollutant degradation yields
thereby increasing the lifetime of charge registered at 120min. irradiation time and kNB
carriers. Doping of TiO2 with metal ion values for undoped and doped TiO2 are
introduces a new energy level (dopant impurity presented in Table 1.
level) by the dispersion of metal nanoparticles in
Dopant presence in the catalyst matrix assures oxidation state and electronic configuration.
enhancement of TiO2 photocatalytic activity, the According with our previous studies, metals are
values of pollutant degradation rate constants are presented in the oxidation state M3+, whose ionic
higher than those obtained in the presence of radii are closer to Ti4+ionic radius in comparison
undoped titania. At the same dopant content, the with M2+. This data is confirmed by small
rate constants vary as function of dopant type, differences between unit cell volume registered
photocatalytic activity generally decreasing in for undoped titania and 0.5-2wt. % metal doped
the following order Fe - doped TiO2 > Co - TiO2 [35]. Among dopants, Fe3+ ion which
doped TiO2 > Ni - doped TiO2. Various dopants presents only 5 electrons on d orbitals is
induced different modification on catalyst probably more accessible to electron trapping
photoactivity depending on their ionic metal than Co3+ or Ni3+ ions (with 6 and 7 electrons)
which have less available half-occupied d Doping of TiO2 with metals not only enhanced
orbitals. The enhanced photocatalytic activity of the separating efficiency of photo-induced
Fe - doped titania compared with Co or Ni- electrons and holes, but also increases the visible
doped titania is revealed by the NB degradation light absorption due to shortening of band gap.
efficiencies obtained at 120 min. irradiation and The absorbance spectra for the three catalysts
2wt. % dopant content, which are 71.77; 67.93 doped with (experimentally set) optimum metal
and 61.48%, respectively. concentrations, were measured at room
Beside dopant type, other parameter which temperature with a Helios UV/VIS
modifies the photocatalytic activity is dopant spectrophotometer and comparisons have been
content. As shown in table1, each dopant made with undoped titania, as shown in Figure
presents an optimum concentration level which 3.
assures the higher NB degradation efficiency. The undoped TiO2 spectrum shows an
According to experimental results the optimum absorption onset at 390 nm, which is in good
concentration for Fe dopant is 0.5wt.% and agreement with the intrinsic energy band gap of
1wt.% in the case of Co and Ni dopants, titania at 3.2eV. The absorption spectra for all
corresponding kNB values being 2.5x10-4 and doped catalysts are shifted to larger wavelength
2.2x10-4 s-1. Dopant level increase up to (red shift). The higher absorbance in the range
optimum concentration positively affected the of 400-600nm (visible light region) is observed
photocatalytic activity. In the case of Co and Ni for (0.5wt.%Fe) – TiO2 catalyst, which confirms
dopants, enhancement of metal content from 0.5 its superior photocatalytic activity on NB
to 1wt.% assures increasing of kNB with 45 and degradation in aqueous solution under UV-VIS
21%, respectively. An opposite behaviour is irradiation.
observed when metal level exceeds optimum
content. kNB values are decreasing with 18; 24; The influence of photocatalyst loading
and 38% for Fe, Co, and Ni, respectively, in the In order to determine the effect of catalyst
case of twice higher content than optimum loading, a series of experiments were carried out
concentration. In accordance with our results, on NB solution with 2.52x10-4 M content and
other authors reported the improved pH=7, by varying the (0.5wt.%Fe) - TiO2photo
photocatalytic activity of 0.5wt.% Fe-doped catalyst dose between 50-500mg/L. From
TiO2, which showed drastically increases of determined pollutant concentration at various
charge-carrier lifetime to minutes and even irradiation times (60- 240min.) the NB
hours beside undoped titania (average lifetime of degradation efficiency and rate constant to each
an electron/hole pair is about 30ns) [43-45]. applied catalyst dose was calculated.
0.40
0.35 TiO2Ni
TiO2Fe
0.30
TiO2Co
Absorbance
0.25 TiO2
0.20
0.15
0.10
0.05
0.00
200 300 400 500 600 700
nm
Fig. 3. UV-VIS absorption spectra of heavy metal doped and undoped TiO2
The results, presented in table 2, show that experiments the optimum catalyst loading of
degradation efficiency increases to almost 99% 250mg/L was applied.
by increasing the catalyst dose from 50 to 250
mg/L. Increase of catalyst loading above The influence of initial pH
250mg/L, leads to degradation rate decrease. The effect of pH on NB photocatalytic
Thus, 79% degradation efficiency was observed degradation was evaluated over the range 4–10,
at 500mg/L dose. The enhancement of into solution with 2.52x10-4 M pollutant content,
degradation efficiency with catalyst loading was at optimum photocatalyst dose (0.5wt.%Fe -
due to the increases in the surface area of the TiO2 = 250mg/L) for irradiation time between
catalyst available for NB adsorption and 60-240 min. The NB degradation efficiencies
degradation. and rate constants for several initial pH
But higher values than optimum catalyst dose conditions are presented in Table 3. The
(250mg/L) also increases solution opacity degradation efficiency increases from 94.4 to
leading to decreases in the penetration depth of 99% as the pH value is incremented from 4 to 7
light into suspension and decreasing the and then decreases under 90% with the
degradation rate [46, 47]. In all subsequent increased value of pH from 7 to 10.
Table 2. The influence of photocatalyst loading on NB degradation efficiency and rate constant
Photocatalyst dose, mg/L kNBx104, s-1 *NB, %
50 1,35 85,71
100 2,50 97,22
250 3,05 98,80
500 1,08 78,96
* irradiation time = 240min
The pH of the solution has complex effects on interaction with charged catalyst. Less or
the photocatalytic oxidation reaction, depending non-polar compounds as it is the case of NB will
on pollutant type and zero-point charge (pHZPC) be better adsorbed on uncharged catalyst surface
of catalyst which in the case of Fe-doped TiO2 is and their degradation rate achieve maximum
around 7. When doped catalyst is dispersed in value near the catalyst ZPC [48]. Therefore, the
water, the first step is hydration of catalyst acidic or alkaline pH does not favour the
surface followed by protonation (pH < pHZPC) or adsorption of NB on the Fe-TiO2 particles.
deprotonation (pH >pHZPC) depending on According to results presented in Table 3, the
solution pH. Thus, the photocatalytic best pH value for NB photocatalytic degradation
degradation of the ionisable organic compounds under mentioned condition is 7, similar data
will be strongly affected by the pH due to their being reported by other authors [49].
pH=7, for different initial NB concentrations only the experimental points during the first 30
between (0.37-8.45) x 10-4 M were performed. min. irradiation. The plot of 1/r0 = f (1/C0)
The r0 values were independently obtained from showed in Figure 5 attests that Langmuir-
kinetic curves by the linear feet, using the Hinshelwood model describes NB
pseudo-first order kinetics of NB degradation at photodegradation which occurs at catalyst
various initial pollutant concentrations are surface. From the intersection of this straight
presented in Figure 4. line with the ordinate results 1/kr and the
Eq. (13) was linearized by its reciprocal calculated value of the reaction rate is kr=1.23 x
expression: 10-7 Ms-1. The adsorption equilibrium constant
1
1
1 Kads=2022 M-1 was calculated from the slope.
r k k K C
0 r r ads 0 (14)
-4
[NB]0=0.37x10 M
6
-4
5 [NB]0=0.67x10 M
ln([NB]0/[NB]t
4 -4
[NB]0=2.52x10 M
2 -4
[NB]0=8.45x10 M
0
0 50 100 150 200 250
t, min
1.2x10
8 kr=0.12 x 10-6Ms-1
Kads= 2022 M-1
8
1.0x10
7
8.0x10
1/r0, M-1s
7
6.0x10
7
4.0x10
7
2.0x10
0.0
0 1x10
4 4
2x10
4
3x10
1/c0, M-1
presented in Figure 6 the presence of EDTA- electron reduction of oxygen towards superoxide
2Na and BQ do not affected NB degradation, radical formation. This disadvantage is
while i-PR drastically decreased the pollutant overcome by the presence of Fe species on the
conversion efficiency. This suggests that surface of catalyst which served as electron-
hydroxyl radical (•OH) are the main active trapping centers for the multi-electron reduction
species involved in NB degradation. Taking into of adsorbed oxygen. Consequently, upon UV-
consideration trapping experimental results but VIS irradiation electrons in the VB of catalyst
also the presence of oxygen vacancy defects are first transferred to the Fe species (redox
(VO) and Fe species on the catalyst surface we potential of Fe3+/Fe2+=0.771V), which act as co-
try to propose a possible mechanism of catalysts, and subsequently are consumed via the
photocatalytic NB degradation on Fe doped multi-electron oxygen reduction process. The
TiO2 under UV-VIS irradiation. It is well known holes left in the VB may react with water and
that delay/retardation of charges recombination generate •OH radicals that are further involved
has a positive effect on photocatalytic activity of in pollutant oxidation. The efficient charge
TiO2. carriers’ separation achieved by Fe doped TiO2
One method applied in this respect is trapping of assures an enhanced catalyst activity on
separated charges like electron reduction of pollutant degradation under UV-VIS irradiation,
oxygen. due to the synergistic effect of the presence on
In the case of oxygen-deficient TiO2 catalysts, the catalyst surface of Fe species and oxygen
like ours Fe doped TiO2, the reduction of vacancies. This is in accordance with results
adsorbed oxygen is inhibited due to VO defects reported by other authors for Fe species-grafted
states localize at 0.78-1.1eV below the on TiO2 catalyst applied in the β-naphtol
conduction band (CB) minimum of TiO2 degradation under visible light irradiation [53].
increases the reaction barrier for the single
1.0
0.9
0.8
0.7
[NB]t/[NB]0
0.6
0.5 i-PR
0.4
0.3
BQ
0.2
EDTA 2Na
0.1
No scavenger
0.0
0 60 120 180 240
Time, min
In addition, the pollutant photocatalytic and some of its degradation intermediates (4-
oxidation could be inhibited when the excess CP, 4-AP, DCAB) concentrations decrease,
H2O2 reacts with oxidative holes on catalyst emphasizing that these are subsequently
surface, as equations11,12 show [54, 58]. Based involved in secondary reactions;
on presented experimental results it was found - an opposite behavior is registered in the case
that optimum H2O2 concentration is 1 x 10-3M, of A and 4-CNB which are accumulated in
which assures after 180 minutes irradiation, the system because these are hardly oxidized
99.99 % pollutant degradation and 88 % organic in the tested working conditions;
chlorine mineralization. - the evolution of DCAB and 4-CNB
The identification of intermediates and 4-CLA concentration confirmed that the second one
degradation mechanism is the degradation product of halogenated
Several organic intermediates like aniline (A), 4- diazobenzene derivate;
aminophenol (4-AP), 4-chlorophenol (4-CP), 4- - after 60 min of irradiation, 4-AP
chloronitrobenzene (4-CNB), 4,4’- concentration was almost neglectable
dichloroazobenzene (DCAB) were identified compared with 4-CP, suggesting that
during 4-CLA photocatalytic degradation in halogenated derivate is hardly degraded; in
UV/TiO2/H2O2 system, by GS-MS method. addition, 4-CP concentration is almost
In order to obtain information on subsequent constant between 60 and 120 min showing
intermediates degradation we compared the that its generation rate is higher than its
specific peak area (directly related with subsequently degradation.
compound concentration) of each GC-MS Based on previously presented results it was
identified byproduct presented in total proposed a reaction sequence for the generation
chromatogram at different irradiation time and of the main intermediates identified during the
the following aspects have been observed: photocatalytic degradation of 4-CLA as it is
- increase of irradiation time leads to 4-CLA presented in Figure 7.
2
. NH Cl
Cl NH NH Cl
OH
Cl N N Cl (DCAB)
OH
2 Cl NO2 (4-CNB)
NH2
OH -NO2- carboxylic
OH acids
OH OH
OH Cl O O C2 C6
Cl -
(4-CP) (benzoquinone)
Cl
HOH OH
(4-CLA) -HN4+
-HCl
NH2 OH
CO2 + H2O
-Cl (4-AP)
OH
NH2
(A)
Fig. 7. Proposed photocatalytic degradation pathways of 4-CLA
The proposed mechanism presents four possible group, with lower steric hindrance compared
pollutant degradation routes: with ortho position; photohydrolisys of 4-CLA
- HO• radicals attack on target pollutants with with 4-AP formation, is a difficult reaction due
replacement of the amino group and 4-CP to HO• radicals and chlorine have similar
formation; electronegativity which inhibiting substitution of
- hydrogen abstraction with anilinyl-radical halogen atom by hydroxyl radicals;
formation, that is subsequently stabilized by The secondary intermediates (4-CP, 4-AP) are
dimerization to DCAB; The attack of HO• hydroxilated [63,64] and further oxidized to
radical on diazogroup of DCAB leads to 4- benzoquinone [65]. This one is subsequently
CNB which is further transformed to 4-CP as converted to other degradation products like
other researchers also reported [59, 60]; muconic, maleic, fumaric and oxalic acids by
- Cl- release with A formation by a several ring cleavage under HO• radical attack [66]. The
steps heterolytic mechanism consisting of final degradation products include Cl-, NH4+,
hydrogen and chloride ions liberation, as NO3- resulted in various steps of proposed
HCl, followed by second hydrogen atom degradation routes, as is presented in Fig. 9. The
elimination with nitrene formation. This one formation of non-chlorinated intermediates like
is subsequently rearranged into seven A, 4-AP agrees with experimental results which
members’ carbene, which is converted to show Cl- releasing as HCl, during irradiation.
aniline [60,61].
- A is oxidized to 4-AP by HO• radicals CONCLUSIONS
addition to the aromatic ring, in accordance H2O2 assisted photolysis and photocatalysis in
with other studies [62]. Since the nature of various systems like UV-VIS/Fe-TiO2 or UV-
HO• radical is highly photolytic electrophilic, VIS/TiO2/ H2O2 are efficient methods for
it preferentially attacks the position with degradation of chloro and nitroaromatic
highest electron density of aromatic ring. pollutants. In proper working conditions
In the case of aniline, the hydroxylation take pollutants advanced degradation (η > 99.6%) is
place in para positions with respect to amino obtained:
•
UV/H2O2 system: [CB]0 = 1.8x10-3M, pH = OH radicals attack to double bound of aromatic
7, [H2O2] = 5.4x10-2M, tirr = 60 min; ring with hydroxylated intermediates formation,
UV-VIS/Fe-TiO2 system: [NB]0 = 0.47x10- followed by their oxidation to benzoquinone,
4 which can be subsequently converted into
M, pH = 4, [Fe-TiO2] = 250mg/L, tirr = 240
min; carboxylic acids through ring cleavage.
UV-VIS/TiO2/H2O2 system: [4CLA]0 = Inorganic ions like Cl-, NO3-, NH4+ are also
1.4x10-3M, pH = 4, [TiO2] = 200mg/L, formed as mineralization products. Results
[H2O2] = 1x10-3M, tirr = 180 min; obtained recommended studied advanced
degradation processes as performant alternative
The degradation pathway for any of studied treatment methods of wastewater with toxic
advanced degradation systems includes initial pollutants content.
REFERENCES
1. BECERRA-CASTRO, C., LOPES, A. R., VAZ-MOREIRA, I., SILVA, E. F., MANAIA, C.
M., NUNES. O. C., Environ. Int., 75, 2015, p. 117.
2. FAURÈS, J.M., HOOGEVEEN, J., WINPENNY, J., STEDUTO, P., BURKE, J., Food and
Agriculture Organization of the United Nations (FAO), Rome, 2012.
3. BIXIO, D., THOEYE, C., DE KONING, J., JOKSIMOVIC, D., SAVIC, D., WINTGENS,
T., MELIN. T., Desalination, 187, 2006, p. 89.
4. DEBLONDE, T., COSSU-LEGUILLE, C., HARTEMANN P., Int. J. Hyg. Environ. Health,
214, 2011, p. 442.
5. GEISSEN, V., MOL, H., KLUMPP, E., UMLAUF, G., NADAL, M. VAN DER PLOEG,
M., VAN DER ZEE, S. E. A. T. M., RITSEMA, C. J., Int. Soil Water Conserv. Res., 3,
2015, p. 57.
6. RASHED, M.N., Organic pollutants - monitoring, risk and treatment. InTech Open, Eds:
Rashed, M.N., 2013, p. 174.
7. MACHULEK, A., OLIVEIRA, S.C., OSUGI, M.E., FERREIRA, V.S., QUINA, F.H.,
DANTAS, R.F., OLIVEIRA, S.L., CASAGRANDE, G.A., ANAISSI, F.J., SILVA, V.O.,
CAVALCANTE, R.P., GOZZI, F., RAMOS, D.D., DA ROSA, A.P., SANTOS, A.P., DE
CASTRO, D.C., NOGUEIRA. J.A., Organic pollutants - monitoring, risk and treatment,
InTech Open, Eds: Rashed, M.N., 2013, p.152.
8. FILIPE, O. M. S., MOTA, N., SANTOS, S. A. O., DOMINGUES, M. R. M., SILVESTRE,
A. J. D., NEVES, M. G. P. M. S., SIMOES, M. M. Q., SANTOS, E. B. H., J. Hazard.
Mater., 323, 2017, p. 250.
9. MALATO, S., MALDONADO, M. I., OLLER, I., ZAPATA. A., Emerging compounds
removal from wastewater: natural and solar based treatments, Springer, Eds: Lofrano G.,
2012, p. 59–76.
10. MALATO, S., FERNANDEZ-IBANEZ, P., MALDONADO, M. I., BLANCO, J.,
GERNJAK. W., Catal. Today, 147, 2009, p. 1.
11. ARQUES, A., AMAT.A. M., Emerging compounds removal from wastewater: natural and
solar based treatments. Springer, Eds: Lofrano G., 2012, p. 77–91.
12. AHMAD, I., AHMED, S., ANWAR, Z., SHERAZ, M. A., SIKORSKI, M., Int. J.
Photoenergy, 2016, 2016, p. 1.
13. MACHULEK, A., QUINA, F.H., GOZZI, F., SILVA, V.O., FRIEDRICH, L.C., MORAES,
J.E., F., Organic pollutants ten years after the Stockholm convention - environmental and
analytical update, InTech Open, Eds. Puzyn, T., Mostrag-Szlichtyng, A., 2012, p. 271-292.
14. OTURAN, M.A., AARON, J.J., Crit. Rev. Environ. Sci. Technol., 44, 2014, p. 2577.
15. BAUER, R., FALLMANN, H., Res. Chem. Intermed., 23, 1997, p. 341.
16. BAXENDALE, J.H., WILSON, J.A., Trans. Faraday Soc., 53, 1957, p. 344.
17. ANDREOZZI, R., CAPRIO, V., INSOLA, A., MAROTTA, R., Catal. Today, 53, 1999, p.
51.
18. GLATZ, W.H., BELTRAN, F., TUHKANEN, T., KANG, J.W., Water Pollut. Res. J. Can.,
27, 1992, p. 23.
19. LIPCZYNSKA-KOCHANY, E., Water Pollut. Res. J. Can., 27, 1992, p. 97.
20. SUNDSTROM, D.W., WEIR, B.A., KLEI, H.E., Environ. Prog., 8, 1989, p. 6.
21. MOZA, P.M., FYTIANOS, K., SAMANIDOU, V., KORTE, F., Bull. Environ. Contam.
Toxicol., 41, 1988, p. 678.
22. WEIR, B.A., SUNDSTROM, D.W., KLEI H.E., Hazard. Waste Hazard., 4, 1987, p. 167.
23. GUITTONNEAU, S., DE LAAT, J., DORE, M., DUGUET, J.P., BONNEL, C., Environ.
Technol. Lett., 9, 1988, p. 1115.
24. DIMEGHANI, M., ZAHIR, O. K., J. Environ. Qual., 30, 2001, p. 2063.
25. ANDREOZZI, R., CAPRI, V., D’AMORE, M.G., INSOLA, A., Ozone. Eng. Sci., 14, 1992,
p. 177.
26. ANDREOZZI, R., INSOLA, A., CAPRI, V., D’AMORE, M.G., Water Res., 26, 1992, p.
639.
27. BOLTON, J.R., LIPCZYNSKA-KOCHANY, E., Environ. Sci. Technol., 26, 1992, p. 259.
28. ONCESCU, T., NIȚOI, I., OANCEA, P., J. Adv. Oxid. Technol., 11, nr. 1, 2008, p. 149.
29. OLLIS, D.F., HSIAO, C.Y., BUDIMAN, L., LEE, C.L., J. Catal., 88, 1984, p. 89.
30. MASTEN, S.J., DAVIES, S.H. R., Environ. Sci. Technol., 28, 1994, p. 180-185.
31. OLLIS, D., AL-EKABI, H., Photocatalytic Purification of Water and Air, Elsevier, Eds.
Ollis, D., Al-Ekabi, H., New York, 1993, p. 164-168.
32. ZHANG, Y., CRITTENDEN, J.C., HAND, D.W., Chem. Ind., 19, 1994, p. 715.
33. RAJESHWAR, K., J. Appl. Electrochem., 25, 1995, p. 1067.
34. PELIZZETTI, E., CARLIN, V., MINERO, C., GRÄTZEL, M., New J. Chem., 15, 1991, p.
351.
35. MALATO, S., BLANCO, J., RICHTER, C., BRAUN, B., MALDONADO, M. I., Appl.
Catal. B, 17, 1998, p. 347.
36. OHNO, T., AKIYOSHI, M., UMEBAYASHI, T., ASAI, K., MITSUI, T., MATSUMURA,
M., Appl. Catal. A, 265, 2004, p.115.
37. LEE, S. Y., PARK, S. J., J. Ind. Eng. Chem., 19, 2013, p. 1761.
38. XU, A.W., GAO, Y., LIU, H.Q., J. Catal., 207, 2002, p. 151.
39. ANPO, M., TAKEUCHI, M., J. Catal., 216, 2003, p. 505.
40. SHEN, X.Z., LIU, Z.C., XIE, S.M., GUOUO, J., Hazard, J., Mater., 162, 2009, p.1193.
41. NI, M., LEUNG, M.K.H., LEUNG, D.Y.C., SUMATHY, K., Renew. Sust. Energy Rev.,
107, 2007, p. 401.
42. NITOI, I., OANCEA, P., RAILEANU, M., CRISAN, M., CONSTANTIN, L., CRISTEA, I.,
J. Ind. Eng. Chem., 21, 2015, p. 677.
43. CORDISCHI, D., BURRIESCI, N., DÁLBA, F., PETRERA, M., POLIZZETTI, G.,
SCHIAVELLO, M., Solid State Chem., 56, 1988, p. 182.
44. BALLY, A.R., KOROBEINIKOVA, E.N., SCHMID, P.E., LEVI, F., BUSSY, F., J. Phys.
D Appl. Phys., 31, 1998, p. 1149.
45. GENNARI, F.C., PASQUEVICH, D.M., J. Mater. Sci., 33, 1998, p.1571.
46. ROBERT, D., DONGUI, B., WEBER, J.V., J. Photochem. Photobiol., A, 156, 2003, p. 195.
47. DANESHVAR, N., SALARI, D., KHATAEE, A.R., J. Photochem. Photobiol., A, 162,
2004, p. 317.
48. SAFARI, M., NIKAZAR, M., DADVAR, M., J. Ind. Eng. Chem., 19, 2013, p.1697.
49. BATHTKANDE, D.S., KAMBLE, S.P., SAWANT, S.B., PANGAKAR, V.G., J. Chem.
Eng., 102, 2004, p. 283.
50. GUETTAI, N., AIT AMAE, H., Desalination, 185, 2005, p. 439.
51. EVGHENIDOU, E., PYTIANOS, K., POULIOS, I., Appl. Catal. B, 30, 2005, p. 259.
52. CRISAN, M., MARDARE, D., IANCULESCU, A., DRAGAN, N., NITOI, I., CRISAN, D.,
VOICESCU, M., TODAN, L., OANCEA, P., ADOMNITEI, C., DOBROMIR, M.,
GABROVSKA, M., VASILE, B., App. Surf. Sci., 455, 2018, p. 201.
53. KONG, L., WANG, C., WAN, F., ZHENG, H., ZHANG, X., Appl. Surf. Sci., 396 ,2017, p.
26.
54. CHU, W., CHOY, W. K., SO, T.Y., J. Hazard. Mat., 141, 2007, p. 86.
55. CHOY, W.K., CHU, W., Chem. Eng. J., 136, 2008, p.180.
56. WATANABE, N., HORIKOSI, S., KAWABE, H., SUGIE, Y., ZHAO, J., HIDAKA, H.,
Chemosphere, 52, 2003, p. 851.
57. NIȚOI, I., OANCEA, P., CRISTEA,I., CONSTANTIN, L., NECHIFOR, GH., J.
Photochem. Photobiol., A, 298, 2015, p. 17.
58. WONG, C.C., CHU, W., Chemosphere, 50, 2003, p. 981.
59. KUMAR, A., MATHUR, N., J. Colloid. Interface Sci., 300, 2006, p. 244.
60. ISHIKAWA, S., BABA, K., HANADA, Y., UCHIMURA, Y., KIDO, K., Bull. Environ.
Contam. Toxicol., 42, 1989, p. 65.
61. OTHEMEN, K., BOULE, P., J. Photochem. Photobiol. A: Chem., 136, 2000, p. 79.
62. CANLE, M., SANTABALLA, J.A. VULIET, E., J. Photochem. Photobiol., A, 175, 2005, p.
192-200.
63. ISHIDA, T., ABE, H., NAKAJIMA, A., KAYA, K., Chem. Phys. Lett., 170, 1990, p. 425.
64. DUNNIVANT, F.M., SCHWARZENBACH, R.P., MACALADY, D.L., Environ. Sci.
Technol., 26, 1992, p. 2133.
65. BANDARA, J., MIELCZARSKI, J.A., LOPEZ, A., KIWI, J., Appl. Catal. B, 34, 2001, p.
321.
66. WINARO, E.K., GETOFF, N., J. Biosci., 57, 2002, p. 512.
Occurrence of Pharmaceuticals and Disinfectants in the Dissolved Water Phase of the Danube
River and Three Major Tributaries from Romania
1National Research and Development Institute for Industrial Ecology ECOIND Bucharest, 71-73 Drumul Podul Dambovitei, 060652,
Bucharest, Romania
2University Politehnica Bucharest, Faculty of Applied Chemistry and Materials Science, 1-7 Polizu Str., 011061, Bucharest,
Romania
*correspondence author (e-mail): [email protected]
The presence of pharmaceuticals in the aquatic ecosystem has received great attention from the
scientific community in the last decades, due to their potential impact on living organisms. This
paper presents a short review of the results of investigations performed by INCD-ECOIND
concerning the occurrence of 32 pharmaceutical compounds belonging to important therapeutic
classes and 2 disinfectants along the Danube River and its tributaries. Grab water samples were
collected from multiple points along the River (10 sites) and from 2 locations for each of the
tributaries Jiu, Olt and Argeș, upstream and downstream of large cities. All samples were
quantified via solid phase extraction, followed by liquid chromatography-tandem mass
spectrometry (LC-MS/MS). Results of these studies show that various pharmaceutical compounds
are present in the dissolved water phase of the Danube River and its tributaries in low to moderate
amounts with variation due to season and location.
In recent years, pharmaceutical compounds have trimethoprim and erythromycin [3,4]. Animal
been recognized as a new class of contaminants and fish farms, where antibiotics are
for water resources, with adverse effects on administered as feed additives and for
aquatic and terrestrial life. therapeutic purposes, are also sources of drug-
The presence and concentration levels of resistant bacteria [5]. Nowadays there are
pharmaceuticals in natural waters require numerous pathways through which
investigation in order to assess their potential to pharmaceutical compounds reach the aquatic
disturb the equilibrium of the aquatic environment (depicted in Figure 1). Effluents of
environment. Thus, a continuous exposure of the municipal wastewater treatment plants
bacterial fauna, even to very low concentrations (WWTPs) are a major source of continuous
of antibiotics, can lead to the persistence of introduction of numerous human pharmaceutical
resistant bacterial strains [1]. Also, the exposure compounds in natural waters. Unused or expired
of aquatic organisms to endocrine disrupting drugs not properly landfilled are also a potential
chemicals, for instance estrogen hormones can source of contamination of surface waters.
affect the reproduction and development of these Moreover, runoffs from agricultural fields,
living organisms [2]. Some studies reported that livestock from farms and fish ponds may contain
bacteria isolated from treated sewage sludge pharmaceutical residues which end up in the
have shown resistance to several antibiotics aquatic environment [6].
including ampicillin, tetracycline, ciprofloxacin,
Danube River is a large stream receiving high remove organic compounds, such as the
anthropogenic inputs and whose water is at the pharmaceuticals with lower levels of
same time an important source of drinking water concentration, which can be found in
for riparian population, in some cases without wastewaters.
being subjected to any treatment process. In this paper [17] we investigated the occurrence
Therefore, there is a need of data on the and removal of nine pharmaceuticals in the
presence of these contaminants in the Romanian WWTP of Bucharest. The influent and effluent
part of the Danube River basin. The aim of these samples were 24h composites and were
works conducted by INCD-ECOIND and collected using flow proportional samples
summarized in this short review article, was to during a nine-day period in the autumn of 2012.
document the occurrence of selected The compounds investigated in this study were
pharmaceuticals along the Romanian part of the acetaminophen (ACE), caffeine (CAF),
Danube River and assess the contribution of its trimethoprim (TRI), norfloxacin (NOR),
three main tributaries: Jiu, Olt and Argeș. ciprofloxacin (CIP), azithromycin (AZI),
Presence of pharmaceuticals and disinfectants sulfamethoxazole (SMX), erythromycin (ERY)
in WWTP effluents and carbamazepine (CBZ). The concentration
Conventional treatment plants, mostly used in ranges of pharmaceuticals found in influent and
Romania, are not designed to completely effluent samples are presented in Table 2.
ERY and NOR were not detected in the sampling period [21] and the deconjugation of
analyzed samples, probably due to their limited conjugated metabolites during the treatment
usage during the period of investigation. process [22] are among the possible
Excepting CIP and AZI, all compounds were explanations.
ubiquitous in WWTP influent and effluent Another investigation was carried out in the
samples. The most abundant one was caffeine. effluents of municipal WWTP of the City of
This widely used compound was found in Brăila, having the Danube River as natural
effluents in concentrations ranging from 2.82 receiver [23]. Treatments in this WWTP include
µg/L to 11.64 µg/L. In effluent samples CBZ primary settling and biological processing
concentration varied from 0.71 μg/L to1.57 (activated sludge). All samples were grab
μg/L, similar to 1.18 μg/L found in Källby samples and were collected between the 16th
WWTP in Sweden [18]. Similar concentrations and the 20th of November 2015.
of SMX and TRI were observed at WWTPs The studied compounds were diclofenac (DCF),
effluents in Spain [14], these two ketoprofen (KET), piroxicam (PIR), ibuprofen
pharmaceuticals being used in the same drug. (IBU), naproxen (NPX), hydrochlorothiazide
Mean removal rates of the compounds (HCT), furosemide (FUR), triclosan (TCS),
determined in this work, calculated as the triclocarban (TCC), and gemfibrozil (GEM).
difference between 9 days mean concentrations The results showed that all effluent samples
of each compound in the influent and effluent contained pharmaceuticals and disinfectants.
samples are depicted in Figure 2. Only IBU was not detected in any of the
ACE and CAF had the highest average analysed samples. Maximum concentrations
elimination yields of 83.3% and 81.0%, were determined for PIR, with average
respectively. Low or no removal rates of CBZ concentration of 103 ng/L. TCS and TCC, the
were also previously reported [19,20]. This disinfectants investigated in samples from
organic compound is known as being very effluents were found in average concentrations
persistent to biodegradation, even at low of 15.6 ng/L and 14.1 ng/L, respectively.
concentrations. An increase in CBZ, TRI and
SMX concentrations in the effluent samples was Occurrence of pharmaceuticals in surface
observed, a phenomenon also reported in other water of Danube River and tributaries
studies [20-22]. In several investigations conducted in INCD-
The fluctuation of the concentrations during the ECOIND since 2012, the presence and fate of 32
Romanian Journal of Ecology & Environmental Chemistry ● No.1 ● 2019
www.incdecoind.ro
58
J. Petre et. al.: Occurrence of Pharmaceuticals and Disinfectants in the Dissolved Water Phase of the Danube River
and Three Major Tributaries from Romania
Fig. 3. Map of the sampling sites in the Danube River basin. S = sampling site
Table 3. Geographic location with GPS coordinates and description of sampling sites.
Sampling point River GPS coordinates
S1- Bazias Danube 44°47'32.61"N; 21°23'20.07"E
S2- Gura Vaii, upstream Drobeta Turnu Magurele Danube 44°40'7.40"N; 22°33'10.74"E
S3- Calafat Danube 43°57'50.94"N; 22°54'15.72"E
S4- Filiasi, upstream Craiova city Jiu 44°34'8.32"N; 23°27'18.14"E
S5- Podari, downstream Craiova city Jiu 44°15'18.48"N; 23°47'25.08"E
S6- Rast, upstream of the confluence with Jiu river Danube 43°51'24.84"N; 23°17'18.79"E
S7 -Bechet, downstream of the confluence with Jiu river Danube 43°45'11.32"N; 23°56'30.69"E
S8 -Downstream Slatina city Olt 44°23'29.63"N; 24°21'4.84"E
S9 -Izbiceni, upstream of the confluence with Danube Olt 43°48'41.71"N ; 24°42'27.71"E
S10 -Islaz, upstream of the confluence with Olt river Danube 43°42'22.72"N; 24°44'4.01"E
S11- Drobeta, downstream of the confluence with Olt river Danube 43°43'2.69"N; 24°48'56.65"E
S12 -Downstream Pitesti city Arges 44°28'45.25"N; 25°40'47.87"E
S13- Giurgiu, upstream of the confluence with Arges river Danube 43°52'37.50"N; 25°58'49.92"E
S14- upstream of the confluence with Danube river Arges 44° 6'38.09"N; 26°38'15.33"E
S15 -Oltenita, downstream of the confluence with Arges river Danube 44° 3'51.89"N; 26°38'45.49"E
S16 -Calarasi Danube 44° 8'15.69"N; 27°20'8.26"E
DCF 0.8 - 7.2 1.3 - 7.5 0.9 - 5.9 1.9 ± 1.4 3.3 ± 2.6 2.1 ± 1.6 50 100 56
KET 0.9 - 3.6 0.8 - 8.4 2.2 - 12.9 1.8 ± 0.6 2.5 ± 3.8 6.4 ± 4.1 44 87 94
TCS 1.7 - 9.1 0.7 - 7.8 1.9 - 18.4 4.4 ± 4.1 2.7 ± 3.2 2.1 ± 2.0 62 100 50
TCC 0.8 - 6.3 0.8 - 54 0.6 - 4.1 2.4 ± 1.7 3.6 ± 1.8 1.5 ± 1.2 100 100 44
Table 5. Range and median of pharmaceutical concentrations and detection frequency of individual
pharmaceuticals from surface water samples
Compound Range (ng/L) Median ± IQR (ng/L)a Frequency (%)b
Caffeine 28.3 - 128.0 76.4 ± 35.6 100
Carbamazepine 5.4 - 15.4 9.65 ± 6.1 100
Sulfamethoxazole 3.2 - 15.7 7.9 ± 2.4 87
Clarithromycin 1.2 - 23.2 3.0 ± 2.7 75
a
Trimethoprim 1.3 - 11.1 2.6 ± 1.2 69 IQR-interquartile range
b
Percentage of particular
Cephalexin 5.6 -17.8 9.9 ± 4.6 50
pharmaceuticals in 16
Ciprofloxacin 3.6 - 4.8 3.7 ± 0.6 19 sampling sites
Figure 5 shows the scatter plots of individual respectively. The fairly regular occurrence of
pharmaceutical concentration versus sampling SMX and TRI in the collected sample is
points on Jiu, Olt and Argeș rivers. As depicted consistent with the compounds occurrence in the
in the figure below, the most contaminated upper Tennessee River basin [32] and Douro
samples were collected from tributary rivers River estuary (Portugal) [33]. Their high degree
downstream of big cities like Craiova (Jiu of persistence in natural waters is explained by
River), Slatina (Olt River) and Bucharest (Argeș their low values of log Kow of 0.91(TRI) and
River). 0.89 (SMX) [34]. Similar concentrations of TRI
CAF and CBZ were also detected in this and SMX were found in the surface waters of
investigation in all surface water samples in Canal Danube-Tisa-Danube from Serbia [35]
concentrations which varied from 28.3 to 128 and in the surface waters of the Henares-Jarama-
ng/L and from 5.4 to 15.1ng/L, respectively. Tajo River system (Madrid, Spain) [36].
Taking into account their ubiquitous occurrence Fluctuations of pharmaceutical concentrations in
in WWTP effluents, CAF and CBZ are the Danube River waters were observed
considered markers of surface water pollution downstream of the Argeș river discharges, the
[30]. most contaminated tributary. TRI, SMX, CLA
The CBZ anticonvulsant, found in the surface and CEF, which were not detected upstream of
waters of Romanian Danube River basin, had the Argeș river discharge, occurred downstream
similar concentrations to those reported for of the confluence in concentrations ranging from
surface waters in Serbia (6-130 ng/L) [31]. The 1.3 ng/L to 15.9 ng/L. Estrogen hormones levels
high frequency detection of CBZ in river waters in surface water of Danube River and the three
is explained by its low removal rate in WWTPs tributaries were tested during a sampling
and by its high resistance in the aquatic campaign carried out in October 2015 using a
environment [31]. newly developed LC-ESI (-)-MS/MS method
SMX and TRI, usually used in combination with automated SPE extraction [26]. The
pharmaceutical formulations, are among the investigated compounds were: Equilin (Eq),
antibiotics most frequently detected in surface Estrone (E1), 17α-Estradiol (αE2), 17β-Estradiol
waters, in concentrations of 69% and 87% (βE2), 17α-Ethinylestradiol (EE2), Estriole (E3).
Fig. 6. Estrogen hormones levels (ng/L) detected along the Danube River
Fig. 7. Estrogen hormones levels (ng/L) detected in Jiu, Olt and Argeș tributaries
ACKNOWLEDGEMENTS. The work has been funded by The Program Nucleu 13N/2009, Project
code: PN 09-13 01 09.
REFERENCES
1. OBST, U., SCHWATZ, T., VOLKMANN, H., Int J Artif Organs, 29, no.4, 2006, p. 387.
2. LIU, S., YING, G.-G, ZHAO, J.-L., CHEN, F., YANG, B., ZHOU, J., LAI, H.-J., J
Chromatogr. A, 1218, 2011, p. 1367.
3. COSTANZO, S.D., MURBY, J., BATES, J., Mar. Pollut. Bull., 51(1-4), 2005, p. 218.
4. DOLEJSKA, M., FROLKOVA, P., FLOREK, M., JAMBOROVA, I., PURGERTOVA, M.,
KUTILOVA, I., CIZEK, A., GUENTHER, S., LITERAK, I., J. Antimicrob. Chemother., 66,
2011, p. 2784.
5. HARNISZ, M., GOŁAŚ, I., PIETRUK, M., Ecol. Indic., 11, 2011, p. 663.
6. GRACIA-LOR, E., CASTIGLIONI, S., BADE, R., BEEN, F., CASTRIGNANÒ, E.,
COVACI, A., GONZÁLEZ-MARIÑO, I., HAPESHI, E., KASPRZYK-HORDERN, B.,
KINYUA, J., LAI, F.Y., LETZEL, T. LOPARDO, L., MEYER, M.R., O'BRIEN, J.,
RAMIN, P., ROUSIS, N.I., RYDEVIK, A., RYU, Y., SANTOS, M.M., SENTA, I.,
THOMAIDIS, N.S., VELOUTSOU, S., YANG, Z., ZUCCATO, E., BIJLSMA, L., Environ.
Int. 99, 2017, p. 131.
7. ***Official Journal of the European Union, 7.6.2018, no. L 141, p. 9.
8. CASTIGLIONI, S., BAGNATI, R., FANELLI, R., POMATI, F., CALAMARI, D.,
ZUCCATO, E., Environ. Sci. Technol. 40, 2006, p. 357.
9. LOGANATHAN, B., PHILLIPS, M., MOWERY, H., JONES-LEPP, T.L., Chemosphere,
75, 2009, p. 70.
10. WATKINSON, A.J., MURBY, E.J., COSTANZO, S.D., Water Res., 41, 2007, p. 4164.
11. LACEY, C., MC MAHON, G., BONES, J., BARRON, L., MORRISSEY, A., TOBIN, J.M.,
Talanta, 75, 2008, p. 1089.
12. SZYMANSKA, U., WIERGOWSKI, M., SOLTYSZEWSKI, I., KUZEMBO, J.,
WIERGOWSKA, G., WOZNIAK, M.K., Microchem J, 147, 2019, p. 729.
13. KOSMA, C.I., LAMBROPOULOU, D.A., ALBANIS, T.A., Sci. Total Environ. 466-467,
2014, p. 421.
Romanian Journal of Ecology & Environmental Chemistry ● No.1 ● 2019
www.incdecoind.ro
64
J. Petre et. al.: Occurrence of Pharmaceuticals and Disinfectants in the Dissolved Water Phase of the Danube River
and Three Major Tributaries from Romania
14. GROS, M., RODRÍGUEZ-MOZAZ, S., BARCELÓ, D., J. Chromatogr. A, 1292, 2013, p.
173.
15. MADUREIRA, T.V., BARREIRO, J.C., ROCHA, M.J., ROCHA, E., CASS, Q.B.,
TIRITAN, M.E., Sci. Total Environ., 408, 2010, p. 5513.
16. GOLOVKO, O., KUMAR, V., FEDOROVA, G., RANDAK, T., GRABIC, R.,
Chemosphere, 111, 2014, p. 418.
17. PETRE, J., IANCU, V.I., VASILE, G.G., ALBU, F., NICULESCU, M., NICULAE, A.C.,
CRUCERU, L., NICOLAU, M., SGEM 2013 Conference Proceedings, ISBN 978-619-
7105-04-9, vol I, 2013, p. 71.
18. BENDZ, D., PAXEUS, N.A., GINN, T.R., LOGEC, F.J., J. Hazard. Mater., 122, 2005, p.
195.
19. ZHOU, J.L., ZHANG, Z.L., BANKS, E., GROVER, D., JIANG, J.Q., J. Hazard.
Mater.,166, 2009, p. 655.
20. JOSS, A., KELLER, E., ALDER, A.C., GÖBEL, A., MCARDELL, C.S., TERNES, T.A.,
SIEGRIST, H., Water. Res., 39, 2005, p. 3139.
21. ZHANG, Y., GEIΒEN, S.-U., GAL, C., Chemosphere, 73, 2008, p. 1151.
22. GULKOVSKA, A., LEUNG, H.W., SO, M.K., TANIYASU, S., YAMASHITA, N.,
YEUNG, L. W.Y., RICHARDSON, B. J., LEI, A.P., GIESY, J.P., LAM, P.K.S., Water
Res., 42, 2008, p. 395.
23. PETRE, J., GALAON, T., IANCU, V.I., CRUCERU, L., NICULESCU, M., J Environ Prot
Ecol, 17, no 1, 2016, p. 119.
24. T., PETRE, J., IANCU, V.I., STANESCU, E., SGEM 2015, Conference Proceedings, ISBN
978-619-7105-40-7, Book 5 Vol. 2, 2015, p. 301.
25. PETRE, J., GALAON, T., IANCU, V.I., VASILE G.G., STANESCU, E., PASCU, L.F.,
SIMION, M., CRUCERU, L., Rev. Chim. (Bucharest), 67(8), 2016, p. 1436.
26. GALAON, T., PETRE, J., IANCU, V.I., CRUCERU, L., VASILE G.G., PASCU, L.F.,
LEHR, C.B., Revista de Chimie, 67(8), 2016, p. 1474.
27. LOOS R., LOCORO G., CONTINI S., Water Res., 44, 2010, p. 2325.
28. ESTEBAN S., GORGA M., PETROVIC M., GONZALEZ-ALONSO S., BARCELO D.,
VALCARCEL Y., Sci. Total Environ., 466-467, 2014, p. 939.
29. KASPRZYK-HORDERN B., DINSDALE R.M., GUWY A.J., Water Res., 42, 2008, p.
3498.
30. AYDIN, E., TALINLI, I., Chemosphere, 90, 2013, p 2004.
31. GRUJIĆ, S., VASILJEVIĆ, S., LAUŠEVIĆ, T., M., J. Chromatogr. A, 1216, 2009, p 4989.
32. CONLEY, J. M., SYMES, S. J., SCHORR, M. S., RICHARDS, S. M., Chemosphere, 73,
2008, p 1178.
33. MADUREIRA, T. V., BARREIRO, J. C., ROCHA, M. J., ROCHA, E., CASS, Q. B.,
TIRITAN, M. E., Sci. Total. Environ., 408, 2010, p 5513.
34. BENDZ, D., PAXÉUS, N. A., GINN, T. R., LOGE, F. J., J. Hazard. Mater., 122, 2005, p
195.
35. PETROVIĆ, M., ŠKRBIĆ, B., ŽIVANČEV, J., FERRANDO-CLIMENT, L., BARCELÓ,
D., Sci. Total. Environ., 468-469, 2014, p 415.
36. FERREIRA DA SILVA, B., JELIC, A., LÓPEZ-SERNA, R., MOZETO, A.A., PETROVIC,
M., BARCELÓ, D., Chemosphere, 85, 2011, p 1331.
37. AGA, D.S., Fate of Pharmaceuticals in the Environment and in Water Treatment Systems,
CRC Pres, London, 2008, p. 300.
38. BOGI, C., SCHWAIGER, J., FERLING, H., MALLOW, U., STEINECK, C., SINOWATZ,
F., KALBFUS, W., NEGELE, R.D., LUTZ, I., KLOAS, W., Environ. Res., 93, 2003, p.
195.
39. GORGA, M., PETROVIC, M., BARCELO, D., J. Chromatogr. A, 1295, 2013, p. 57.
National Research and Development Institute for Industrial Ecology ECOIND Bucharest, 71-73 Drumul Podul Dambovitei, 060652,
Bucharest, Romania
*correspondence author (e-mail): [email protected]
The present article presents the expertise realized by the Department of Environmental Monitoring
Pollution Evaluation within the INCD ECOIND, in the evaluation of the quality of urban soils in
the municipality of Bucharest and the main big cities in Romania. The current data available at the
level of the 27 member states of the European Union show that annually over 100,000 hectares of
land are introduced into the urban environment, a direct consequence of the development of cities.
There are a number of legislative obstacles to strategic soil protection measures. Moreover, at the
level of the local authorities there is a conflict regarding the measures of soil protection in the long
term, on the one hand, and, the accelerated economic development in the short term, on the other.
European environmental experts consider that the urban development, absolutely necessary for the
economic growth, requires an adequate management of the natural resources in order for the
development to be done on a sustainable basis, respectively to follow a series of strategic
objectives. In our country, at least in the last decade, we find on a large scale the conversion of
industrial areas into commercial or residential areas. The footprint of industrial activities can be
found even after long periods of time present by identifying the remnant of soil pollution or in those
areas known as historically polluted (for example the town of Copsa Mica).
The conclusions stemming from the assessment of pollution in urban areas over large areas, in
correlation with the potential sources of pollution, underline the need to monitor the quality of soils
in the urban environment, but also to apply a performance management in order to protect this
natural resource in the long term.
The activities presented in this article aimed at Urban soils accumulate different types of
the level of the main cities in Romania: Iasi, pollutants from different sources of pollution.
Timisoara, Cluj Napoca, Constanta, Craiova, The development of cities in Romania has
Galati, Brasov, Ploiesti and Oradea to highlight undergone major changes in the last decades,
the quality of the soils in the urban environment. whether we are talking about the rehabilitation
Urban ecology studies show that soil pollution is of the existing infrastructure or the construction
especially important because it is directly related of new infrastructure, or the restriction of
to public health. The increase of the population industrial areas and, very important, their
density in the urban environment is felt by the conversion into commercial or residential areas.
influence of the anthropic activities and on the Soil is at the same time a very important natural
quality of the soils (mainly the industrial resource that must be kept under sustainable
activities). It is found that urban soils differ conditions for future generations.
greatly from natural ones, respectively from Figure 1 shows the cities in Romania that were
those not found under the direct influence of the investigated within the project for evaluating the
anthropic factors in the big cities. quality of urban soils.
Soil management is a priority to protect this influences, industrial pollution sources in each
resource so important, especially in the urban city studied, land use and functionallity, etc);
environment where the anthropic pressures are - GPS localizations of the samples and maps of
very high. It is absolutely mandatory to take into the experimental field for each city.
account the relationships between soil and
human health, as well as between soil and B. Laboratory activities:
climatic conditions, respectively the adaptation - relevant quality indicators determined by
to climate changes of the last decades. modern analytical techniques (ICP-MS);
- adequate test methods, standardized and
EXPERIMENTAL PART validation of the results using Certified
The location of study areas on the maps are References Materials.
presented in Figure 1. Activites was conducted Figure 4 shows schematically the activities
in two main directions: carried out for investigating the quality of urban
A. Field activities: soils.
- dimensioning the number of samples to soil to It is noted that the in terrain documentation
be sampled with adequate equipment for activities occupy the first place, especially
sampling; important, because the office documentation
- obtaining relevant information and details cannot supplement the updated information’s
(microrelief, hydrographic network, climatic obtained on the site.
The involvement of local public authorities with environmental assessment who make a series of
competences in ensuring the quality of the recommendations and monitoring proposals for
environmental conditions in urban areas is future stages, practically a contribution and a
defining and necessary. That is why the final support for the realization of management on
report of the obtained results is directed to them, soil resources in the urban environment.
it is a processed information by specialists on
Fig .5. Pollution in Copsa Mica city related to actual potential source
ACKNOWLEDGEMENTS. This work was carried out through the “Program Nucleu”, financed
by the Ministry of Research and Innovation of Romania, project codes: PN 16 25 02 10 and
PN 16 25 02 06.
REFERENCES
1. U.N. 2008. World urbanization prospects: the 2007 revision. New York, NY: United
Nations.
2. ANTROP, M., Landscape Ecol., 15, 2000, p. 257–270
3. BOLUND P., HUNHAMMAR S., Ecol. Econ., 29, 1999, p. 293–301
4. MCBRATNEY, A., FIELD, D.J., KOCH, A., Geoderma, 213, 2014, p. 203–213.
5. SPARLING, G.P., SCHIPPER, L.A., J. Environ. Qual., 31, 2002, p. 1848–1857
6. SPARLING, G., SCHIPPER, L., Agric. Ecosyst. Environ., 104, 2004, p. 545–552.
7. SPARLING, G.P., SCHIPPER, L.A., BETTJEMAN, W., HILL, R., Agric. Ecosyst.
Environ., 104, 2004, p. 523–534.
8. BRISTOW, K.L., MARCHANT, S.M., DEURER, M., CLOTHIER, B.E., Proc. 19th World
Congress of Soil Science, Brisbane, Australia, 2010.
9. HUBER, S., VERNIK, T., SIEBIELEC, G., VRSCAJ, B., Soil in the City. Urban Soil
Management Strategy, Book, 2012,
10. *** Order no. 756 of November 3, 1997 for approving the Regulation regarding the
assessment of environmental pollution
Danube Delta: monitoring and ecological status. A link between the past and the future
Romania
*corresponding author (e-mail): [email protected]
The paper aims a review of the water quality monitoring and evaluation studies conducted at
National R&D Institute for Industrial Ecology over the last ten years, at the level of aquatic
ecosystem complexes in the Danube Delta.
The paper will include a broad compendium of data including: a data bank of abiotic variables
which control the structure and composition of biotic communities; studies of ecological status
assessment; methodology development for assessing the bioaccumulation of chemical pollutants
(metals) in the biota; ecotoxicological study of surface water and sediment on aquatic organisms
(algae, crustaceans, rotifers); studies on the benthic invertebrate species as biological vectors for
pathogens; technical / scientific support for 5 national / international project proposals; 19 papers
published in international / national journals; 22 papers presented at international / national
scientific conferences.
DNA metabarcoding. The development of DNA E 29ᵒ04’749”), S7) Murighiol (km 64) (N
sequencing technologies has been a promising 45ᵒ02’426”; E 29ᵒ11’090”), S8) Uzlina (km 70)
alternative for biodiversity monitoring, (N 45ᵒ04’452”; E 29ᵒ13’384”), S9) Ivancea (km
especially as these techniques can allow rapid 16) (N 44ᵒ58’219”; E 29ᵒ27’829”), S10) Sfantu
taxonomic identification, high accuracy, cost- Gheorghe Port (N 44ᵒ53’467”; E 29ᵒ35’663”),
effective analysis allowing multiple samples to S11) Black Sea Confluence (km 0) (N
be analyzed simultaneously by contrast with 44ᵒ53’098”; E 29ᵒ36’655”). The geographical
conventional methods, higher costs and time localization of the sampling sites was performed
analysis as well as the vast and specific using GPS type system map 60CSx-Garmin
knowledge on groups of organisms (Figure 1).
(identification and quantification of Data bank. Physico-chemical quality elements
phytoplankton, phytobenthos and benthic such as pH, conductivity, dissolved oxygen
invertebrates) [8, 9]. The use of specific DNA (DO), total dissolved solides (TDS), chemical
sequences to identify species can overcome the oxygen demand (COD), biochemical oxygen
problems mentioned above. In other words, demand (BOD), heavy metals (Fe, Mn, Zn, Cd,
these alternative methods have the ability to Cr, Cu, Ni, Pb, Hg, As), amonium (N-NH4),
fundamentally change the assessment of the nitrates (N-NO3), nitrites (N-NO2), total nitrogen
ecological status of freshwater systems (TN), phosphates (P-PO4), total phosphorus
worldwide. (TP), chlorides, sulphates, sulphides, anionic
In the present paper, the main results of Danube surfactants, calcium, magnesium, sodium,
and Danube Delta systems water quality petroleum products, polycyclic aromatic
obtained after more than ten years monitoring hydrocarbons (PAH) (12 compounds), phenolic
period were presented. compounds, organochlorine pesticides (α-HCH,
ß-HCH, -HCH, -HCH, heptaclor, aldrin,
EXPERIMENTAL PART dieldrin, endrin, DDT/DDD/DDE), triazine
Study area. The monitoring program was pesticides (atrazin, simazin), ureic pesticides
developed at spatial (Danube-Danube Delta (diuron, isoproturon, monolinuron) were
ecological system) and temporal level (monthly characterized in water samples according to
data over ten years’ period). specific EN ISO standard methods. Alongside
The main sampling sites were selected on the chemical indicators detected in water samples,
Pontic Ecoregion along Danube: S1) Isaccea numerous parameters such as As, Cd, Cr, Co,
(km 103) (N 45ᵒ17’051”; E 28ᵒ26’956”, (RO14 Cu, Hg, Ni, Pb, Zn, Fe, Mn, PAH (12
typological category) and Danube Delta (RO15 compounds), organochlorine pesticides (α-HCH,
typological category): S2) Tulcea Upstream (km ß-HCH, -HCH, -HCH, heptaclor, aldrin,
82) (N 45ᵒ11’757”; E 28ᵒ47’372”), S3) Tulcea dieldrin, endrin, DDT/DDD/DDE),
Downstream (km 70) (N 45ᵒ11’492”; E polychlorinated biphenyls (PCBs) and benzene,
28ᵒ48’733”); S4) Nufaru (km 101) (N toluene, ethylbenzene and xylene (BTEX) were
45ᵒ09’111”; E 28ᵒ55’099”), S5) Baltenii de Sus characterized in sediment samples.
(km 97) (N 45ᵒ06’752”; E 28ᵒ59’212”, S6)
Mahmudia (km 90) (N 45ᵒ04’685”;
S1
S2 S3
S4
S5
S6 S8
S7
S9
S10
S11
0
Fig.1. The geographical localization of sampling sites along Danube: (Isaccea (S1) and Danube
Delta: Tulcea Upstream (S2), Tulcea Downstream (S3), Nufaru (S4), Baltenii de Sus (S5),
Mahmudia (S6), Murighiol (S7), Uzlina (S8), Ivancea (S9), Sf. Gheorghe Port (S10), Black Sea
Confluence (S11)
Biological quality elements such as values were compared with the reference area
phytoplankton and bacterioplankton (total and values (with little or no anthropic impact or
faecal coliforms, streptococci) as well as benthic historical data) and the subsequent classification
invertebrates and bacteriobenthos were detected in ecological states (very good, good, moderate,
both in water and sediment samples. weak, bad).
Ecological status assessment. Abiotic Ecotoxicological study. The toxicity of water
(temperature, oxygenation, mineralization, and sediment samples was evaluated using
acidification, nutrients, pollution with priority / various aquatic organisms such as: algae
priority hazardous substances) and biotic (Selenastrum capricornutum), crustaceans
characteristics (diversity, species presence, (Daphnia magna, Heterocypris incongruens)
numerical abundance / biomass) were and rotifers (Brachionus calyciflorus).
considered for assessing the ecological status of The ecotoxicological study performed along
Danube and Danube Delta aquatic ecosystems Danube Delta freshwater samples was done
following the WFD and RBMP requirements. during 2013. The methods used and the tests
Briefly, the ecological status assessment of the conditions are described in Table 1.
studied freshwater systems based on benthic The Danube Delta freshwater toxicity was
invertebrates’ fauna involved the organisms classified according to the hazard classification
sampling considering the standard methods, the system for natural water described by Persoone
drawing up of the taxonomic groups list, the et al. (2003) [11]: Class I – no acute hazard, no
morphological identification and the abundance test did not reveal toxic effect; Class II – slight
estimation. The resulting data were used for acute hazard, 20%≤ effect percentage <50%, in
various indices calculation (such as at least one test; Class III – acute hazard, 50% ≤
Ephemeroptera-Plecoptera-Trichoptera index, effect percentage <100%, in at least one test;
Shannon-Wiener diversity index, family number Class IV – high acute hazard, effect percentage
index, Oligochaeta-Chironomidae index, = 100%, in at least one test; Class V – very high
functional groups index, water flow preference acute hazard, effect percentage =100%, in all
index and multimetric index). The indexes tests.
Romanian Journal of Ecology & Environmental Chemistry ● No.1 ● 2019
www.incdecoind.ro
74
C. Stoica et. al.: Danube Delta: monitoring and ecological status. A link between the past and the future
Methodology for assessing the metals were collected in May and October of 2015.
bioaccumulation in benthic invertebrates. The Benthic invertebrates assemblages (live
bioaccumulation of numerous metals (such as: specimens) were sampled using Van Veen
As, Se, Sb, Cd, Cr, Cu, Co, Fe, Mn, Mg, Ni, Pb dredge (sampling depth of 10 cm).
and Zn) was quantified from two bivales species The samples for microbiological parameters
and one gasteropod species collected during analysis were collected in 1 L glass bottles (for
summer and autumn 2013 from S7 and S8. The water) and 400 g (for sediment). The recipients
applied methodology was detailed in Gheorghe were properly sterilized at 160˚C. Benthic
et al. (2017) [12]. macroinvertebrates were separated by taxonomic
Benthic invertebrates as biological vectors for groups using Motic stereomicroscope and
pathogens. S6, S7, S8 and S10 sampling sites identified by specific determination keys. The
were selected along Danube Delta for the identification and characterization of
identification of potential vectors of pahogens in microorganisms was carried out on the
the benthic communities’ structure. The samples OmniLog automated system (Biolog Inc., USA).
Table 1. Bioassays used to assess the water and sediment toxicity in Danube Delta (according with
Gheorghe et al. (2016) [10]
Method / Tested Examined Test duration/ Sample
Test type Treatment
Microbiotest organisms behavior temperature matrix
OECD 201 Selenastrum Acute / Growth 72h, Undiluted
Water
AlgaltoxKit FTM capricornutum Chronic inhibition 21-25oC
OECD 202 Acute
Daphnia magna Mortality 24-48h, 20oC Water Undiluted
DaphtoxKit FTM
ASTM Standard Guide
24 h,
E1440-91 Brachionus calyciflorus Acute Mortality Water Undiluted
25oC
RotoxKit FTM
Mortality
ISO 14371 Heterocypris 6 days,
Chronic Growth Sediment Untreated
OstracodtoxKit FTM incongruens 25oC
inhibition
the level of the ecosystem complexes Gheorghe Branch sites, due to the intake of
represented by the Danube and the Danube phytoplankton from lakes and canals. It is the
Delta (Sfantu Gheorghe Branch), were followed case of S8 (Uzlina) sampling site which
by profound changes in the taxonomic recorded the highest numerical densities during
composition and distribution of the biocenoses. 2003-2013 (40.145 ind/L).
The densities of both coliform bacteria and The benthic invertebrate fauna was composed of
Enterococcous in water samples were high species belonging to Gasteropoda, Bivalvia,
during the summer and autumn sampling Oligochaeta, Insecta, Crustacea taxonomic
campaigns. Moreover, various antibiotic groups, with greater abundance and diversity in
resistant bacteria were identified [22]. However, the stations located on Sfantu Gheorghe Branch,
annual averages of faecal bacteria densities compared to those on the Danube likewise
decreased in recent years in Danube Delta, due phytoplankton communties, but lower compared
to the reduction of nutrient concentration (N, P) to those reported by other studies conducted on
[23, 24]. The qualitative analysis of Danube [26, 27, 28, 29, 30, 31, 32, 33]. This
phytoplankton community along the Danube reduction of the taxonomic richness from
Delta sampling sites over ten-years study, upstream to downstream was considered to be
emphasized the presence of species belonging to due to the heterogeneity and size of the particles
the classes Baccilariophyceae, Chlorophyceae, / sediment granulation, alongside the increase of
Euglenophyceae and Cyanophycae. In all control factors pressure [34].
sampling sites, the oligo-betamezosaprobe, The heavy metal profile in the Danube Delta
betamezosaprobe, beta-alfamezosaprobe and sediment had a specific configuration. The
alfamezosaprobe species dominated. The concentration of heavy metals (Zn, Hg, Cu)
taxonomic diversity of phytoplankton included increased until 1989, due to the increase of
64 species of algae, but the most frequent were industrialization in Central and Eastern Europe
Asterionella formosa, Cymatopleura solea, [35], including in Romania. However, during the
Melosira varians, Navicula gracilis, Cymbella last 5 decades a decrease of metals concentration
ventricosa, Phacus longicauda, Scenedesmus in the Danube Delta was recorded (Table 2).
quadricauda, Pediastrum duplex, Amphipleura Although, the concentration of metals in the
pellucida, Fragilaria crotonensis, Nitzschia sediment diminished in the last 10 years
sigmoidea [25]. The distribution of between 2 to 5 folds, however, some of them
phytoplankton species was conditioned by the have been adsorbed on the surface of the debris
availability of light, dissolved oxygen, nutrients, or humic acids. The neglecting of plans
but also by the dynamics of the hydrological regarding intensive agriculture in the Danube
regime. The phytoplankton densities increased Delta, contributed to the reduction of Cd
from Isaccea (S1) (32.750 ind/L, average concentration as this compound is part of
numerical density calculated during 2012-2013) fertilizers.
located on the Danube downstream to the Sfantu
Table 2. Variation of heavy metals (mg / kg s.u) in the sediments of the Danube Delta during the
last five decades
1950 2003-2009
Quality element 2012-2013
U.M Winkels et al., 1998 [35] Vosniakos et al., 2010 [18]
Cu 38 14.8-194 4.65-45.9
Pb 36 7.5-51.3 4.76-41.3
Zn mg/kg d.m 90 29.8-218 17.7-93.1
Cr 50 7.5-61.9 7.62-32.5
Ni 56 19-111 10.8-49.8
Cd <0.5 <0.5-1.5 BDL*
*BLD-below detection limit
Decreased Pb may also be associated with time, the phasing out of gasoline with Pb is a
changes in land use, as it also includes complementary method of reducing this
fungicides and other pesticides. At the same contaminant. The increase of metals
concentration along Danube Delta sampling more affected by anthropic and environmental
points was due to the nonpoint contamination pressure factors rather than ‘chemical quality
from non-ferrous deposits abandoned after the elements’ [41].
transitions in the Romanian economy [36]. In Ecological status assessment.
addition, PAH concentrations increased almost Characterization of the state variables (both
seven-fold compared to concentrations chemical and biological quality elements) that
determined in 1950 [35], confirming the influence the composition and structure of
negative impact of navigation development (data macroinvertebrates organisms, as well as the
not shown). dynamics of these biological elements, allowed
Moreover, the organochlorine pesticides the assessment of the ecological status of the
concentration decreased after 1990, with Danube and Danube Delta. Therefore, taking
changes in the political sector, but still persist in into account the "worst case" principle and the
the sediments of the Danube Delta. A visible results obtained during 2012-2013 from both
decrease between 2012-2013 and 2003-2011, biological and chemical quality elements, the
with the exception of DDT and its degradation assessment induced „good” ecological status for
products was observed [15, 16, 17, 18, 25, 37]. Danube system (typological category RO14) and
However, there was a positive correlation "moderate" ecological status for Danube Delta
between the concentrations of the (typological category RO15) (data not shown).
organochlorine pesticides and the benthic The assessment of freshwater systems ecological
indices: Saprobic index (IS), Oligochaeta- status was completed by an in-vitro
Chironomidae (OCH/O) and Hilsenhoff biotic ecotoxicological study.
index (IBH) at S6 (Mahmudia), S7 (Murighiol), Ecotoxicological study. Primary producers
S9 (Ivancea) and S10 (Sf. Gheorghe) in terms of (algae, Selenastrum capricornutum) and
increase values of the indices due to the consumers (planktonic crustaceans, Daphnia
increased values of organochlorine pesticides magna and rotifers, Brachyonus calyciflourus as
(data not shown). well as benthic crustaceans (Heterocypris
As a result, the structure and composition of the incongruens) were used to evaluate the Danube
benthic communities changed, by reducing the Delta water and sediment quality according to
number of taxonomic groups, the replacing of hazard classification system of natural water
sensitive sepcies with the tolerant ones, often [11]. The water samples showed no acute toxic
invasive as well as the replacing of shredders or effect (Class I) or slightly acute toxic effect
collectors with detritivores (data not shown). (Class II). The sediment samples were more
Studies conducted after 1990 have shown a toxic than water samples due to the pollutants
pronounced simplification of biocenoses accumulation (e.g metals, PAH, organochlorine
integrated into aquatic systems [38,39,25], pesticides). Their main toxicity classes were
preceding a period of restoration of the structure from Class II – slightly acute toxic effect and
and composition of the benthic invertebrate Class III – acute toxic effect up to Class IV –
fauna [40]. high acute toxicity. The largest number of water
Overall, a general tendency of benthic fauna toxic responses was observed in case of
recovery in terms of taxonomic richness was Selenastrum capricornutum algae, followed by
observed during the monitoring period at the Daphnia magna crustaceans and Brachyonus
level of ecosystem complexes in the Danube and calyciflourus rotifers. In case of sediment, the
Danube Delta (data not shown). highest number of toxic responses were obtained
A large database was created based on the from Heterocypris incrongruens ostracods [10].
monitoring of Danube Delta water quality over In addition to the development of the database
ten years’ period (2003-2013) and subsequently, comprising a large number of water quality
it was possible to perform a comprehensive indicators, the study of freshwater systems
study of Danube Delta ecological status ecological status assessment as well as an
assessment. The macroinvertebrates’ community ecotoxicological study, a methodology for the
composition was used as they are closely related evaluation of metals bioaccumulation in biota
to habitat and substrate heterogeneity. was implemented.
Moreover, ‘biological quality elements’ were
Methodology for assessing the metals component of aquatic ecosystems, the group was
bioaccumulation in benthic invertebrates. The largely examined in our over ten-years studies.
researches that founded the above mentioned A number of functions performed by benthic
methodology was carried out in 2013 (July and invertebrates’ fauna have been mainly related to
September) along Danube Delta (S7, S8 and trophic relations (accumulating, transforming
S10) regarding the accumulation of chemical and transferring the stored energy from organic
pollutants (metals) in Mollusks (two Bivalve detritus decomposition and nutrient transfer to
species: Anodonta sp., Unio sp. and one the higher trophic levels, information circulation
Gasteropode: Viviparus sp.) [42]. and systems self-regulation [45,46, 47, 48]. The
The results pointed out that the metals involved advantage of using benthic invertebrates’ fauna
in the metabolic processes (Fe, Mn, Zn, Cu and in the freswater quality evaluation studies was
Mg) have a greater accumulation capacity than due to their ability to maintain in their structure
the toxic ones (Pb and Cd). This fact was also the effects of environmental pressure [49, 40].
explained by the presence of these metals in Moreover, the invertebrates have relatively long
high concentrations, reported in previous studies life and widespread distribution, limited
[43]. Moreover, Vasile et al. (2006, 2005) [20, mobility and they cannot avoid adverse
44] observed that Pb was either mobile or bound conditions when exposed to environmental
to manganese oxides and organic matter in issues (such as eutrophication, acidification, low
sediment samples. The mobile Pb concentration oxygen concentrations, hydromorphological
was higher in the sediment at S7 due to the changes or other problems combined with
increase of the total concentration of metals in habitat loss (e.g. embankments, etc.).
the sediment, compared to the concentration of Thus, the benthic communities were considered
metals detected at S8, where a decrease suitable for either the bioaccumulation of
concentration in the mobile form for Cu, Zn and specific toxic pollutants in their structure, or
Pb was detected in 2004, compared to 2003. characterization and assessment of ecological
Furthermore, Vosniakos et al. (2010) [18] status of aquatic ecosystems quality, including
showed that the mobile fraction (1.2-3.1 mg / the identification of various benthic taxonomic
kg) of Cu represents 41%, and Ni showed a groups as vector for pathogens.
relatively low mobility compared to its total Benthic invertebrates as biological vectors for
concentration. pathogens. The identification of potential
Gheorghe et al (in tech) highlithed that the pathogen vectors of the benthic communities’
bioaccumulation level fluctuated considering the structure was carried out in S6, S7, S8 and S10
species, metals type, and sampling sites. A sampling sites along Danube Delta. The samples
greater difference of bioaccumulative metals were collected in May and October of 2015.
impact (Ni and Zn) at S7 compared to S8 was The results pointed out that alongside the
observed. The study of metal bioaccumulation structure of benthic invertebrate fauna (potential
capacity in the mollusks shell from Danube vectors for pathogens) from the ecological
Delta aquatic system underlined that metals system of the Danube Delta, a wide range of
involved in metabolic processes (such as Fe, stable endogenous bacteria were identified.
Mn, Zn, Cu, and Mg) had a greater storage Since the frequency of occurrence reported in
capacity than the toxic ones (such as Pb and Cd) 2014 was greater than 50% (Figure 2) [40],
[12]. The selectivity of the metal several benthic invertebrates’ groups were used
bioaccumulation was represented as follows: as sentinels to assess the prevalence of
Fe> Mn> Zn> Cu> Pb> Co> Cd, in case of pathogens microorganisms, such as: filters
Viviparus sp. shell, while the shell of Anodonta (Dreissena polymorpha, Anodonta cygnea, Unio
sp. had a higher accumulation capacity of metals pictorum), scrapers (Viviparus acerosus,
(Cu, As, Cr, Zn) compared to Unio sp. (data not Esperiana esperi, Esperiana acicularis,
shown). Overall, bioaccumulation factors were Lithoglyphus naticoides, Theodoxus danubialis
subunitary, which indicated a slowly and Chironomidae) and detritivores
bioaccumulation process along Danube Delta. (Oligochaeta, Ceratopogonidae).
Since, the macroinvertebrates represent the key
Although 15 pathogen species were detected in Moreover, benthic invertebrates’ fauna was
all sampling sites subjected to untreated strongly related with pathogens and can be
wastewater discharges, mainly linked with further used as alternative monitoring systems
animal and human fecal waste, Citrobacter for waterborne contamination. In addition, since
freundii and Raoultella plancticola / the Danube Delta water body was framed in
ornithinolytica were the most common bacteria 2013 as „moderate” ecological status according
species, especially associated with Diptera with WFD, alternative methods needs to be
larvae and Mollusks species. tested in order to meet the good ecological status
by 2027.
CONCLUSIONS
The paper presented a summary of all the studies Future recommendations. Since the good
carried out in the National R&D Institute for ecological status for all water bodies by 2015
Industrial Ecology for more than 10 years at the was not achieved, alternative methods needs to
level of aquatic ecosystems complexes of be tested to meet this criterion by 2027. Several
Danube and Danube Delta, considering the steps further were made at international level
anthropogenic pressure after the industrialization [50] by means of DNA metabarcoding
period. The studies aimed to highlight the identification of prokaryotic and eukaryotic
ecological integrity of aquatic systems organisms, yet certain "methodological
considering both chemical and biological limitations" [51] have been identified, namely:
components monitoring. The main issues that a) biomass estimation [52]; b) detection of rare
Danube and Danube Delta still facing are: high species [53]; c) incomplete reference libraries
organic load (expressed COD and BOD), [54, 4].
pathogenic microbial load as well as, the Considering those issues, our future studies will
increase of antibiotic bacterial resistance, high focus on the application of alternative methods
concentrations of heavy metals (copper, nickel, at national level, which may contribute to the
zinc, mercury), organochlorine pesticides identification of functional diversity based on
(lindane, DDT/DDE/DDDs), PAHs and PCBs, gene expression (transcription), and thus, to
mosly detected in sediment. A reduction of meet a WFD target that could not be adequately
sensitive benthic invertebrates’ species and covered based on conventional morphological
replacing with the tolerant ones was observed. identification methods.
ACKNOWLEDGEMENTS. This work was financially supported by the National Core Program
within several contracts: 20N/2019, Project no. PN 19- 04 02 01, 13N/2009, Projects no. 09-13 02
03, PN 09-13 02 14, 12N/2006, Project no. PN 06-12 01 13.
REFERENCES
1. CORDIER, T., LANZEN, A., APOTHELOZ-PERET-GENTIL, L., STOECK, T.,
PAWLOWSKI, J., 2019, Trends in Microbiol., 27, nr. 5, p. 387-397.
2. EC, 2000, Directive 2000/60/EC of the European Parliament and of the Council of 23
October 2000 establishing a framework for the Community action in the field of water
policy, Off J, L 327, p. 1-72.
3. POIKANE, S., ZAMPOUKAS, N., BORJA, A., DAVIES, S.P., VAN DE BUND, W.,
BIRK, S., Environ. Sci. Policy, 44, 2014, p. 237-246.
4. HERING, D., BORJA, A., JONES, I., PONT, D., BOETS, P., BOUCHEZ, A., BRUCE K,
DRAKARE, S., HÄNFLING, B., KAHLERT, M., LEESE, F., MEISSNER, K., MERGEN,
P., REYJOL, Y., SEGURADO, P., VOGLER, A., KELLY, M., Water Res., 138, 2018, p.
192–205.
5. HERING, D., BORJA, A., CARSTENSEN, J., LAURENCE, C., ELLIOTT, M., FELD,
C.K., HEISKANEN, A.S., JOHNSON, R.K., MOE, J., PONT, D., SOLHEIM, A.L., VAN
DE BUND, W., Sci. Total Environ., 408, 2010, p. 4007-4019.
6. VOULVOULIS, N., ARPON, K.D., GIAKOUMIS, T., Sci. Total Environ., 2017, 575, p.
358–366
7. MOLDOVEANU, M., RÎŞNOVEANU, G., Conferinţa Ştiinţifică Jubiliară a Institutului
Naţional de Hidrologie şi Gospodărire a Apelor, 28-30 septembrie 2010, Bucureşti (in
Romanian).
8. LOBO, J., SHOKRALLA, S., COSTA, M.H., HAJIBABEI, M., COSTA, F.O., Sci. Rep., 7,
2017, p. 15618.
9. SERRANA, J, MIYAKE, Y., GAMBOA, M., WATANABE, K., Ecol. Indic., 101, 2019, p.
963-972.
10. GHEORGHE, S., STOICA, C., PAUN, I., LUCACIU, I., NITA-LAZAR, M., CRISTOFOR,
S., J. Environ. Prot. Ecol., 17, no. 1, 2016, p. 171-181.
11. PERSOONE, G., et al., Environ. Toxicol., 18, no. 6, 2003, p. 395.
12. GHEORGHE, S., STOICA, C., VASILE, G.G., NITA-LAZAR, M., STANESCU, E.,
LUCACIU, I., Metals toxic effects in aquatic ecosystems: modulators of water quality, 4, In:
Water Quality, InTech, Hlanganani Tutu (Ed.), ISBN 978-953-51-2882-3, DOI:
10.5772/65744, 2017.
13. ZALASIEWICZ, J., WILLIAMS, M., STEFFEN, W., CRUTZEN, P., Env. Sci. & Tech.,
44, 2010, p. 2228–2231.
14. KELLER, E., BARBARA, M., LATHJA, K., CRISTOFOR, S., Wetlands, 18, no. 1: 1998,
p. 42-50.
15. PETRE, J., CRUCERU, L., NICOLAU, M., VASILE, G., MITRIŢĂ, M., IANCU, V., Rev.
Chim. (Bucharest), 55, no. 4, 2004, p. 264-268.
16. VOSNIAKOS F, PETRE, J., VASILE, G.G., CRUCERU, L., NICOLAU, M., MITRITA,
M., IANCU, V., CRUCERU, I., Fresenius Environ. Bull., 15, no.5, 2006, p. 401–408
17. VOSNIAKOS, F., VASILE, G., PETRE, J., CRUCERU, L., NICOLAU, M., MITRITA, M.,
IANCU, V., CRUCERU, I., Fresenius Environ. Bull., 17, 1, 2008, p. 372–389
18. VOSNIAKOS, F., PETRE, J., PASCU, L.F., VASILE, G., IANCU, V., STANILOAE, D.,
NICOLAU, M., CRUCERU, L., GOLUMBEANU, M., Fresenius Environ. Bull., 19, no. 1,
2010, p. 20–29.
19. VASILE, G., PETRE, J., CRUCERU, L., NICOLAU, M., MITRIŢĂ, M., IANCU, V.,
VOSNIAKOS, F., J. Environ. Prot. Ecol., 8, no. 4, 2007, p. 739-747.
20. VASILE, G., CRUCERU, L., PETRE, J., J. Environ. Prot. Ecol., 7, no. 2, 2006, p. 293-300.
21. LIŠKA, L., WAGNER, F., SLOBODNIK, J., Joint Danube Survey – Final Report ICPDR –
International Commission for the Protection of the Danube River, 2008, p. 235.
22. BANCIU (CATRANGIU), A., NICULESCU, D., NITA-LAZAR, M., LUCACIU, I.,
STOICA, C., MIHAESCU, G., J. Environ. Prot. Ecol., 17, no. 1, 2016, p. 127-135.
23. KIRSCHNER, A.K., KAVKA, G.G., VELIMIROV B., MACH, R.L., SOMMER, R.,
FARNLEITNER, A. H., 2009, Water Res., 43, p. 3673–3684.
24. PÁLL, E., NICULAE, M., KISS, T., ŞANDRU, C.D., J. Med. Microbiol., 62, no. 11, 2013,
1635-1640.
25. STANESCU, E., STOICA, C., VASILE, G., PETRE, J., GHEORGHE, S., PAUN, I.,
LUCACIU, I., NICOLAU, M., VOSNIAKOS, F., VOSNIAKOS, K., GOLUMBEANU, M.,
Environmental Security Assessment and Management of Obsolete Pesticides in South-East
Europe, NATO Science for Peace and Security Series C: Environment Security, L.I.
Simeonov, F.Z. Macaev, B.G. Simeonova (Eds), Springer Science+Business Media
Dordrecht, 21, 2013, p. 229-248.
26. GRAF, W., LEITNER, P., PLETTERBAUER, F., The Danube River Basin, The Handbook
of Environmental Chemistry, Liška (Ed.), Springer-Verlag Berlin, Heidelberg, 2015.
27. BÓDIS, E., TÓTHA, B., SZEKERES, J., BORZA, P., SOUSA, R., Limnologica, 49, 2014,
p.1–9.
28. CSÁNYI, B., IHTIMANSKA, M., PAUNOVIĆ, M., PODANI, J., SZEKERES, J., Proposed
section typology of the Danube River based on macroinvertebrates collected by different
sampling methods, 40 th IAD Conference, 2014.
29. LIŠKA, L., WAGNER, F., SLOBODNIK, J., 2008, Joint Danube Survey – Final Report
ICPDR – International Commission for the Protection of the Danube River, p. 235
30. OERTEL, N., NOSEK, J.N, Proc. 36th International Conference of IAD, 2006, p. 275-279.
31. OERTEL, N., NOSEK, J.N., Internat. Assoc. Danube Res., 2006, p. 36.
32. BERTHOLD, E., KAISER, I., Lauterbornia, 50, 2005, p. 15-17.
33. BERNERTH, H., STEIN, S., Lauterbornia, 48, 2003, 57–60.
34. SOMMERWERK, N., HEIN, T., SCHNEIDER-JACOBY, M., BAUMGARTNER, C.,
OSTOJIC, A., SIBER, R., BLOESCH, J., PAUNOVIC, M., TOCKNER, K., Rivers of
Europe, K. Trockner, C.T. Robsinson, U. Uehlinger (Eds.), London, Burlington (MA), San
Diego (CA): Academic Press, 2009, p. 59-112.
35. WINKELS, H.J., KROONENBERG, S.B., LYCHAGIN, M.Y., MARIN, G., RUSAKOV,
G.V, KASIMOV, N.S., Appl. Geochem., 13, no. 5, 1998, p. 581-591.
36. VIGNATI, D.A.L., SECRIERU, D., BAGATOVA, Y.I., DOMINIK, J., CÉRÉGHINO, R.,
BERLINSKY, N.A., OAIE, G., SZOBOTKA, S., STĂNICĂ, A., 2013, J. Environ.
Manage., 125, p. 169-178.
37. STOICA, C., GHEORGHE, Ş., PĂUN, I., STĂNESCU, E., DINU, C., PETRE, J.,
LUCACIU, I., Romaqua, 1, no. 91, 2014, p. 14-20.
38. RÎŞNOVEANU, G., IGNAT, G., Analele Ştiinţifice ale Institutului de Cercetare Dezvoltare
Delta Dunării, 1993, p. 177-184 (in Romanian).
39. IGNAT, G., VĂDINEANU, A., CRISTOFOR, S., NAFORNIŢA, G., RÎŞNOVEANU, G.,
CIUBUC, C., DINU, C., FLORESCU, C., CRAIOVEANU, M., Analele Ştiinţifice ale
Institutului Delta Dunării, Tulcea, 1993, p. 87-94 (in Romanian).
40. STOICA, C., GHEORGHE, S., PETRE, J., LUCACIU I., NITA-LAZAR, M., Environ. Eng.
Manage. J., 13, no. 9, 2014, p. 2243-2252.
41. STOICA, C., CAMEJO, J., BANCIU, A., NITA-LAZAR, M., PAUN, I., CRISTOFOR, S.,
PACHECO, O.R., LOPEZ, M.G., Water Sci. Technol., 73, no. 10, 2016, p. 2413-2421.
42. STOICA, C., GHEORGHE, S., LUCACIU, I., STANESCU, E., PAUN, I., NICULESCU,
D., Soil Sediment Contam., 23, 2014, p. 1-16,
43. VASILE, G., TĂNASE, I.G., DINU, C., 2010, Anal. Lett., 43, p. 1199-1206.
44. VASILE, G., CRUCERU, L., NICOLAU, M., PETRE, J., IANCU, V., Rev. Chim.
(Bucharest), 56, no 8, 2005.
45. COVICH, A.P., PALMER, M.A., CROWL, T.A., BioScience, 49, no. 2, 1999, p. 119-127.
46. COVICH, A.P., AUSTEN, M.C., BÄRLOCHER F., CHAUVET, E., CARDINALE, B.J.,
BILES, C.L., INCHAUSTI, P., DANGLES, O., SOLAN, M., GESSNER, M.O.,
STATZNER, B, MOSS, B., 54, no. 8, 2004, p. 767-775.
47. REISS, H., KRONCKE, I., Mar. Pollut. Bull., 50, no. 12, 2005, p. 1490-1499.
48. RÎȘNOVEANU, G., CRISTOFOR, S., ADAMESCU, M., CAZACU, C., IGNAT, G.,
PARPALA, L., NĂVODARU, I., MOLDOVEANU, M., TOROK, L., IONICĂ, D.,
ZINEVICI, V., TUDOR, M., PREDA, E., IBRAM, O., VĂDINEANU, A., Volumul
Programului Cercetare de Excelență 2005 – 2008, MENER energie, Editura Universității
Politehnica, 2008, p. 238-247.
49. POP, O.G, GRUIA, R., MARCULESCU, A., Environ. Eng. Manage. J., 9, 2010, p. 1593-
1599.
50. ELBRECHT, V., VAMOS, E.E., MEISSNER, K., AROVIITA, J., LEESE, F., Methods
Ecol. Evol., 8, no. 10, 2017, p. 1265-1275.
51. MAJANEVA, M., DISERUD, O., EAGLE, S., HAJIBABEI, M, EKREM, T.,
Metabarcoding &Metagenomics 2: e26664, 2018.
52. ELBRECHT, V., LEESE, F., PloS one10: e0130324, 2015.
53. LERAY, M., KNOWLTON, N., PeerJ 5: e3006, 2017.
54. HAJIBABAEI, M., BAIRD, D.J., FAHNER, N.A., BEIKO, R., GOLDING, G.B., Philos.
Trans. R. Soc. Lond. B. Biol. Sci. 371, no. 1702, 2016, 20150330.
Edited by EIKON