Current Trends in Oceanography and Marine Science Strezov a and Nikolova E

Current Trends in Oceanography and Marine Science Strezov A and Nikolova E. Curr Trends Oceanogr Mar Sci: CTOMS-103. Research Article DOI: 10.29011/ CTOMS-103.100003 Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae

Alexander Strezov*, Emilia Nikolova Department of Radio Chemistry and Radio Ecology, Sofia University, Bulgaria

*Corresponding author: Alexander Strezov, Department of Radio Chemistry and Radio Ecology, Sofia University, Bulgaria. Email: [email protected] Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003 Received Date: 20 June, 2018; Accepted Date: 03 July, 2018; Published Date: 09 July, 2018

Abstract The ecological status at eight coastal locations (Shabla, Kaliakra, Tuzlata, Ravda, Ahtopol, Sinemoretz and Rezovo) along the Bulgarian Black Sea coast was studied by determining of 32 elements in 121 samples of eight green Macroalgae species Ulva rigida, Ulva lactuca, Enteromorpha, Enteromorpha compressa, Cladophora vagabunda, Cladophora coleotrix, Chaetomorpha gracilis and Bryopsis plumosa during the period 1996-2011. The element concentrations were determined by Energy Dispersive X Ray Fluorescence Analysis (EDXRFA). Results for 19 elements in the three most widespread green algae Ulva rigida, Enteromorpha intestinalis, Cladophora vagabunda, are compared in all the locations along the Bulgarian coast to evaluate the level of accumulated elements. It was shown that Ulva rigida and Bryopsis plumosa species accumulate studied elements less than the other species and the heavy metal concentration in green Macrophytes from Tuzlata, Ravda and Sinemoretz is higher compared to the other locations. The obtained data show that no serious contamination by hazardous elements in green algae along the Black Sea shore during the studied period was found and some of the studied green algae species are suitable for heavy metal assessment in Black Sea marine ecosystems. The obtained results establish a database for element concentration in green Macroalgae and their ecological impact on marine ecosystems along the Bulgarian Black Sea coast.

Keywords: Black Sea; Green Macroalgae; Trace Metals; contamination. The determination of widespread bio monitors will XRFA Method allow comparison of accumulated element concentrations over large areas [4-5]. Introduction The Black Sea is a low salinity basin, half-isolated from the The ecological monitoring of marine ecosystems can be per- Mediterranean and its hydrology and phytobentos concentration formed by controlling marine organisms that have been success- is different from the other seas in the same area. Toxic elements fully used as bio-indicators. The bio monitors are important indi- (and nuclides) are introduced by airborne contaminants, rivers, oil cators for evaluation of various contaminants impact on marine pollution or by direct discharge of industrial wastes into the sea. ecosystems. Macroalgae have been used as indicators for trace ele- Trace elements are transported via the seawater currents, coming ments, because of their accumulation capacity and influence along from the Danube and other rivers. The ecological conditions at the the trophic chaining the marine environment [1-3]. Algae can bio coastal zone are diverse which results in element concentration concentrate amounts of biologically significant elements in their variations in algae depending on many factors - species type, geo- tissues. Some elements are essential for aquatic plant growth and graphical location, season etc. their biological uptake is a function of the free element concen- Green Macroalgae have been extensively used to monitor tration in seawater. However, increase in element concentrations marine pollution in various geographical areas [6-9]. Green algae above certain values can be toxic to marine biota. Some algae have are among the widespread at the Bulgarian Black Sea coast, some mutation capability so they can survive the ecological stress of the of them (Ulva rigida, Enteromorpha intestinalis, Cladophora

1 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003 vagabunda)�� can be ��met ��at almost all areas, ��so ��they are ��appropri- window and 160 eV (KETEK). The determination is performed ate for comparative assessment of contaminants concentration in with three radionuclide sources 55Fe(AMERSHAM) by means of different geographical areas. Green Macroalgae species have been an exciting head, especially designed for low Z elements. In this used as bio-indicators in the Black Sea and other marine environ- case the construction, the distances between the source and sample ments in neighboring seas [10-15]. Trace element concentrations and sample-detector are greatly reduced and optimized, which al- in Black Sea green alga species have been determined by other lows elements with atomic number Z = 13 to 25 to be analyzed. authors [16-19] while data about heavy and toxic metals at the Bulgarian Black Sea coastal zone is scarce and insufficient. The The third system is equipped with SDD detector with 0.7 µm Be window and 170 eV energy resolution at 5.9 KevMN-Kline purpose of this paper is to study the level and accumulation of 32 238 elements in eight macrophytes, collected from nine locations along (KETEK). An exciting head with source of Pu is specially de- the Bulgarian Black Sea coast during the period 1996 - 2011. It is signed for analyzing of elements with Z between 17 and 35. This important to determine whether these algae can be used as bio- head is very suitable for analyzing very low concentrations of these indicators for some element pollution and evaluate the impact on elements. The calibration procedures for each spectrometer are the marine ecosystem habitat. Energy-dispersive X-Ray fluores- based on set of international standards like Bowen’s kale, orchard cence method (EDXRFA) was used for trace element determina- leaves, grass (IAEA - V10), fish flesh (IAEA), copepod (IAEA), tion.[20-21]. This method was applied because of its Multielement Nettle etc. The results were checked with IAEA standard - lichen. capability, nondestructive analysis, simple sample preparation, Lichen was chosen because of the similar matrix with the green relative little time consuming, sufficient sensitivity, accuracy and algae due to their similar biochemistry. The Lab has also taken good reproducibility. Compared to ICP, the amount of analyzed part in two International Inter compression Projects - Worldwide sample is bigger (30 g), which reduces the influence of inhomo- Proficiency Test for X-Ray Laboratories PTXRFIAEA/05 and 06 geneity error. IAEA (2009). The LLD for elements with Z between 13 and 14 is about 1000 µg/g, for Z between 15 and 20 - about 500 µg/g, for Z Materials and Methods between 22 and 56 about 1 to 2 µg/g. The LLD for elements Hg, Pb and U is about 5 µg/g. Sampling sites in this study were selected to cover the whole Bulgarian coastline from north to south - Shabla, Kaliakra, Tuzlata, Results and Discussion Rossenetz, Ravda, Sozopol, Ahtopol, Sinemoretz and Rezovo and to possess abundant algae species. Eight green Macroalgae species There is a dependence between microelement bio sorption (Ulva rigida, Ulva Lactuca, Enteromorpha Enteromorpha�� com� and their function in the organism. Most intensively accumulated pressa, Cladophora vagabunda, Cladophora coleotrix, Chaeto�� are those microelements that play a major part in the metabo- morpha gracilis, and Bryopsis plumosa) were collected during lism. A considerable number of elements form stable complexes the period between spring 1996 and spring 2011.The algae were with proteins, phosphates and lipids. Fe, MN, and Co take part in collected in the subtidal zone at 1 - 4 m depth and those abundant- enzyme formation and as complex compounds participate in ex- ly covered with epiphyte were ejected. The samples were rinsed change reactions. The enriched in elements lipids from algae play in clean seawater, followed by distilled water, dried to constant a major biogeochemical role for the processes of fixation, transport weight (85°C) and the grinded 0.5 grams of three subsamples. and sedimentation in the hydrosphere. Three spectrometric systems are available in EDXRFA Lab. The selected green Macroalgae species are suitable for ap- The first one is equipped with Si(Li) detector with 12 µm Be win- plication because they possess potential qualities to be used as bio- dow and 170 eV energy resolution at 5.9 KevMN-Kαline (PGT). indicators, the algae are sufficiently widespread, characteristic for The data are acquired with a multichannel analyzer (AX-1), inter- the studied areas, possess enough biomass and are easily collected; faced to a personal computer, that applies specialized software for and they react to the environmental changes. 32 elements have data acquisition and spectra processing. This system is combined been measured using EDXRFA in 121 green algae samples - 25 with an exciting head, based on an annular source 241Am (3.7 GBq) for Ulva rigida, 20 for E17 for C. vagabunda, 14 for Ulva lactuca (AMERSHAM) and three types of secondary targets (Mo, Dy, Sn) etc. in the period 1996 - 2011. Data for 19 elements are presented in two excitation modes - transition and reflection in order to select in this paper. The concentrations of some elements are below the the desired exciting energy. For determination of heavy elements, LLD limits of the method and the specific spectrometric systems. which are essential for biology the system uses secondary target The concentrations of the most interesting macro and trace ele- of Dy, that allows detection of elements down to the Z = 50. This ments are presented in tables and graphs. construction permits the measurement of great number of elements with a high sensitivity by achieving the desirable experimental It is interesting to evaluate the impact of elements accumula- conditions of low background level and at a high count rate. tion in a certain algae species in all locations along the Black Sea coast. This evaluation can be done at sites where multiple sam- The second system is equipped especially for light element pling has been performed during the years and where most of most analysis with Si pin diode detector, Peltie cooling, with 7 µm Be abundant algae were collected. For this reason, we have chosen

2 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Ulva rigida, E. intestinalis and C. vagabunda species to assess the ecological status of the coast. The results for 19 detected elements (Figure 1) concentrations in the three most widespread green Macroalgae species from the Bulgarian Black Sea coast, are presented in (Table 1) for Ulva rigida (Figure 2-6) (Table 2) for E. intestinalis (Figure 7-11) and in (Table 3) for C. vagabunda (Figure 12 -16). The measured high element concentrations in algae (2000 - 50000 µg/g) are those for the microelements - Al, Si, P, S Cl, K, Ca, the highest of which are for Cl and S. Cl is essential for the algae metabolism and these high concentrations are understandable. The amount of S in marine algae is much higher, then the terrestrialplants, which is probably due to the dissolved sulfates in the sea water which are sorted by the algae. Fe, Br and Sr concentrations are with one or two orders of magnitude higher than the other elements in all algae species.

Figure 1: Map of sampling locations.

3 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Table 1: Element concentratioin in Ulva rigida.

4 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Ulva Rigida - Fe

1200

1000

800

600 ppm

400

200

0 3 0 0 0 m p 0 3 6 S S m 0 9 6 a a S p p 9 3 h h l lS u 0 4 6 S S a a zS A p 0 9 9 K K u z S p u 9 5 T u z S A p 0 5 1 T u ja v S p 0 1 8 T B a v S p u 9 5 R a v S p 0 2 R a m zA S u 0 5 R o o n A u 0 6 P s r A p 9 5 7 S a e r S u 0 9 0 Fe P a r A p 0 M V a t tS m 6 V h S m 9 0 5 A h t u u 0 0 A h S A m p A t in S h n zS A S i e S R

Figure 2: Fe concentration in Ulva species.

Ulva Rigida - Mn, Cu, Zn

80 Cu 70 Zn Mn 60

50 ppm 40

0 6 9 6 p 9 3 S u 0 4 z A p 0 6 u z S p 9 9 T u z S u 9 5 T u a A p 0 5 T j v S p 0 1 B a v S p 1 8 R a v S u 9 R a A p 5 R m z S 0 2 Mn o o n u 0 5 P s A u 0 6 S a r A p 9 e r u 5 Zn M P a rS 0 7 V a tA p 9 0 V h tS m 0 6 Cu A h tS m 9 0 A h u u 0 5 A tS A m 0 h in S p A S in zS S e R

Figure 3: MN, Cu, Zn concentration in Ulva species.

5 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Ulva Rigida - Sn, Pb

20 Sn Pb

15 ppm

0 6 6 9 9 3 p u 0 4 S p 0 6 9 z A S p 9 9 5 u z z S u p 0 5 T u u a A p 0 1 T T j v vS S p 1 8 B a a v S u 9 5 R a A p 0 2 5 R R m z S u 0 0 6 o o n A u p 9 5 P S s r rA u 0 7 Sn a e a rS p 9 0 M P a tA S 0 6 V V h t m m 9 0 5 A h tS u u 0 0 A h S A m p A t in S S h n z A S i e S R

Figure 4: Sn and Pb concentration in Ulva species.

Ulva Rigida - Sr, Ba

350

300

250 Ba Sr

200 ppm 150

100

0 6 9 6 p 9 3 u 0 4 6 zS A p 0 9 9 z S p u 9 5 u u z S p 0 5 T T u ja A S p 0 1 8 T v v S p 1 9 5 B a a v S u p 0 2 R R a m A S u 0 5 R o z n u 0 6 o s rA A p 9 5 P S e r u 0 7 Ba a a rS A p 9 0 M P V a t S m 0 6 V h t S m 9 0 5 A h t u u 0 0 A h S A m p A t in S h n zS A S i e S R

Figure 5:Sr and Ba concentration in Ulva species.

6 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Ulva Rigida - Br, I

900

800

700

600 I

500 Br ppm 400

300

200

100

0 6 9 6 p 9 3 u 0 4 6 zS A p 0 9 9 u z S p u 9 5 T u z S A p 0 5 T u ja v S p 0 8 T a v S p 9 5 B a v S p 0 2 R a S u 0 5 6 R R m n A u 0 9 o s r A p 5 7 P a e r S u 0 9 0 I M P a r tA p 0 V a tS m 6 V h S m 9 0 5 A h t u u 0 0 A h S A m p A t in S h n zS A S i e S R

Figure 6: Br and I concentration in Ulva species.

7 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Table 2: Element concentration in Enteromorpha intenstinalis.

Enteromorpha Intenstinalis - Fe

3000

2500

2000

Fe 1500 µg/g

1000

500

0 9 9 9 3 8 9 0 3 4 5 1 6 5 3 5 6 2 3 0 3 9 9 9 0 9 9 0 0 0 0 0 9 0 0 0 9 0 0 0 0 p p p p p p p p u p p p p p u u p p S S S m S S A S S m S a lS lS lS z z S z z z lS v s tS tS A A S S z a a a u u z u u u E a o h h in in in z e h K K K u S S S e S T T T T T T K R R A A R R

Figure 7: Fe concentration in Enteromorpha species.

8 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Enteromorpha Intenstinalis - Mn, Cu, Zn

100

50 µg/g Cu 40 Zn Mn 30

0 9 9 9 9 9 p p 9 3 8 S S p 0 9 9 a l S p p 9 0 3 h a l lS S p 0 0 4 S K a a z S m p 0 5 Mn K u z S p 0 1 6 K u z S u 0 5 T T z S A p 9 u u z z S p 0 3 5 T T u u l S p 0 0 6 Cu T T E v S p p 9 2 K a s tS u 0 3 0 R o h tS A u 0 0 3 R h n A p 0 A A i n S m p S i in S S z zS S e e R R

Figure 8: MN, Cu and Zn concentration in Enteromorpha species.

Enteromorpha Intenstinalis - Cd, Sn, Pb

25 Cd Pb 20 Sn

15 ppm

0 9 9 9 9 p 9 9 3 S p p 0 8 9 a lS p 9 9 0 h a lS S p p 0 3 4 a l S S m 0 0 5 S K K a z z p p 0 1 6 Sn K u u zS S u 0 9 5 T T z S A p p 0 3 u u z z S S p 0 5 T T u u l v S p 0 6 2 Cd T T E a s S p 9 0 3 K o t S u u 0 0 3 R h t A A p 0 0 R A h in n S m p A S i n S S i z zS S e e R R

Figure 9: Cd, Sn and Pb concentration in Enteromorpha species.

9 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Enteromorpha Intenstinalis - Br, I

500

450

400

350

300 I 250

µg/g Br 200

150

100

0 9 9 9 9 p 9 9 3 8 S p p 0 9 9 0 a lS S p p 9 0 3 h a l S p 0 4 5 S a l S S m p 0 1 K K a z z S S p 0 0 6 5 K u u z z S u p 9 0 3 T T u u z A p p 5 T u z lS S 0 0 6 2 I T T u E v S p p 9 0 3 T a s tS S u 0 0 3 K R o h t A u p 0 0 R h n A S m p A A i in n S S i zS zS S e e R R

Figure 10: Br and I concentration in Enteromorpha species.

Enteromorpha Intenstinalis - Sr, Ba

1000

900

800

700

600

500 ppm Ba 400 Sr

300

200

100

0 9 9 9 9 p 9 9 3 8 S p p 0 9 9 0 a lS S p p 9 0 3 h a l lS S p 0 4 5 S K a a z S m p 0 0 1 K u z S S p u 0 6 5 K T u z z S p 9 0 3 T u u z A S p p 5 T T u z l S 0 0 6 2 Ba T u E v S p p 9 0 3 T K a s tS S u u 0 0 3 R o h t A p 0 0 R A h in A S m p A in n S S S i z zS S e e R R

Figure 11: Cd, Sn and Pb concentration in Enteromorpha species.

10 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Table 3: Elemennt concentratioin in Cladophora vagabunda.

11 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Cladophora vag - Fe

7000

6000

5000 Fe 4000 ppm 3000

2000

1000

0 9 2 3 6 8 9 5 8 6 5 5 1 9 9 9 9 9 9 0 9 9 0 0 0 p m p p p p u p p p p u S S S S S S A S rS S S A l z z z z z z N a t n z a u u u u u u s s h i e K T T T T T T a T A S R M

Figure 12: Fe concentration in Cladophora species.

Cladophora vag. - Mn, Cu, Zn

450 400 350 300

250 Cu ppm 200 Zn 150 Mn 100 50 0 9 9 2 3 p 9 9 6 Mn S m p 9 8 9 l S S p 9 9 5 a z z S p p 0 8 K u u z S S u 9 6 Cu T T u z z A p 9 5 T u u z S p 0 5 1 T T u N rS p 0 0 T s a tS p u a s h S A T in z M A S e R

Figure 13: MN, Cu and Cu concentration in Cladophora species.

12 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Cladophora vag - Cd, Sn, Pb

ppm Cd 10 Sn Pb 5

0 9 9 2 3 p 9 9 6 Pb lS m p 9 8 9 a S S p 9 9 5 K z z S p p 0 8 u u z S S u 9 6 Cd T T u z z A p 9 5 T u u z S p 0 5 T u S p 0 1 T sN r S p 0 T a sa t S u T h in zA M A S e R

Figure 14: Cd, Sn and Pb concentration in Cladophora species.

Cladophora vag - Ba, Sr

500 450 400 350 300 250 ppm Ba 200 Sr 150 100 50 0 9 2 9 9 3 p 9 6 lS m p 9 8 a S S p 9 9 5 z z S p 9 0 8 K u u z S p u 9 6 Ba T T u z S p 9 5 T u z zA S p 0 5 T u u S p 0 1 T T sN r S p 0 a sa t S u T h in zA M A S e R

Figure 15: Ba and Sr concentration in Cladophora species.

13 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Cladophora vag - Br, I

400

350

300

250

200 ppm I 150 Br 100

0 9 2 9 9 3 p 9 6 lS m p 9 8 a S S p 9 9 5 z z S p 9 0 8 K u u z S p u 9 6 I T T u z S p 9 5 T u z zA S p 0 5 T u u S p 0 1 T T sN r S p 0 a sa t S u T h in zA M A S e R

Figure 16: Br and I concentration in Cladophora species.

The overall Fe mean value obtained for Ulva rigidia 510 – many of the samples are in the range of 5 - 7 µg/g (while the µg/g and the variation range is wide (300 – 1100 µg/g ). The con- mean Zn value is 38 ±3 µg/g) except several samples where the centration of Fe in E. intestinalis is twice higher, compared to Ulva concentrations are three or four times higher. Cu and Zn concen- rigida, the mean value being about 1100 µg/g (except Sizemore trations are almost constant in the uncertainty limit of the method Autumn’96 - 1840 µg/g and Rossenetz Spring’05 - 2885 µg/g). and are also independent of sampling time and location. Two sam- The mean Fe concentration in Cladophora vagabunda is close to pling sites (Bjala and Pomorie) have considerably higher Cu and E. intestinalis with exception of Tzarevo Spring`96 - 6680 µg/g. It Zn concentration. The case with the Rossenetz location (Sping’05 is clear that the two green Cladophora and Enteromorpha species - 111 µg/g) is completely different – there is a copper mine in the accumulate twice higher Fe concentration than Ulva species. vicinity and the concentration of all elements is completely differ- The results for MN in Ulva vary in very narrow limit (the ent than all other sites due to the human mining activity. mean value 70 ± 3µg/g). The same is true for the results MN in E. Copper concentration determined in algae during the intestinalis which are also close (the mean value 70 ± 3µg/g) like present investigation is comparable with those reported by for in Ulva rig. except two samples, (Rezovo Summer’00 – 38 µg/g, algae collected from Crimean coast at the Black Sea, while for Sizemore Spring’03 - 140 µg/g) which means that MN concentra- lead (nonessential element, but it is present in all biomaterials) the tion is almost constant, independent of sampling time and loca- concentrations are lower than those found by Burdin. Pb is found tion. The situation for MN in Cladophora is quite different – in in seawaters mainly in the form of different organic compounds. one third of the samples the concentration is about 50 µg/g, while As it is seen from Pb and Sn data for the three green algae in the in the other samples the concentration is four times higher except highest lead concentration is measured in spring 1996 - 1998, Tuzlata Spring`99 - 371 µg/g and Ahtopol Spring`05 - 430 µg/g. while the concentrations decrease during the next sampling period. The element Cu and Zn belong to the group of biologically important metal ions and their compounds can be found as trace metals The measured concentration for Pb is close to typical Pb obtained in the biosphere. Cu and Zn are essential for organisms’ life and for the terrestrial plants. its toxicity is connected with the concentrations of other essential The concentration of Sn in Ulva are relatively constant elements and some marine algae show an ability to accumulate Cu (mean 11 ± 3 µg/g), except Bjala (28 µg/g). The Sn values for E. and Zn. Trace metals should be monitored because they play an intestinalis vary more and are higher than the other two green al- important role in metabolism and their high or low concentrations gae samples and are close to those measured in marine sediments. can be equally harmful to the living organisms. Cd concentration in Ulva (~4 µg/g) was measured only in four Cu and Zn concentration in Ulva rigida, are presented in samples from Tuzlata (2), St. Constantine & Elena and Ahtopol. The Cu values in Ulva, Enteromorpha and Cladophora are close Cd is not measured in all samples for the other green algae (in

14 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

Enteromorpha - 4, and C. vagabunda only two sites). The overall vagabunda, than to the two green Ulva rigida and Enteromorpha� concentration of Pb, Sn and Cd in the sample Tuzlata Spring 98 is vaga. Special attention should be about the green alga Bryopsis three times higher than all other measured algae samples. plumosa. The elements from alkaline earth group -Sr and Ba are accumulated to much higher extent – two to four orders of mag- It should be pointed out that some samples differ substan- nitude (~30000 µg/g for Ba (~50 µg/g for the other green algae); tially from the others, which can be attributed to the various fac- ~4000 µg/g for Sr (~200 for the others). The concentrations of I tors affecting elements accumulation and the specific conditions is ~2000 µg/g (~50 µg/g for the others). We have measured [13] of studied locations especially depending on the sampling time radio metrically the Ra daughter isotope concentrations and also (mainly spring). Another example for great deviation of elements found three orders of magnitude higher concentration compared to values is the data (µg/g) for Ulva rigida in Bjala location Spring all other algae. This comes to confirm the specific accumulation of 2004 and C. vagabunda in Tzarevo Spring`1996. The concentra- alkaline earth elements by the green alga Bryopsis plumosa. tion of halogen elements Cl, Br and I, which are biologically important for the algae and marine biota, were measured also in Ulva Conclusion rigida, E. intestinalis and C. vagabunda. The obtained average value for Br and I are close for both green algae Ulva rigida, E. The concentrations of 32 elements have been determined in intestinalis (360 µg/g - for Br and 47 µg/g for I. while the Br value eight Black Sea green Macroalgae from nine locations along the in Cladophora is twice lower. Lower values were measured for E. Bulgarian Black sea coast during the period 1996-2011. If Fe data intestinalis from Tuzlata Autumn 2005, Sinemorez Autumn 1996, during the monitoring period are compared, no clear seasonal or Sinemorez Autumn 2002, suggesting seasonal dependence. The location dependence can be observed. It has been shown that Ulva measured Cl values in E. intestinalis and C. vagabunda are higher rigida and Bryopsis plumosa species accumulate studied elements (mean value 30800 µg/g) compared to those of Ulva r. (22400 less than the other species and the element concentration in green µg/g). The values in half of the samples in Cladophora are twice macrophytes from Tuzlata, Ravda and Sinemoretz is higher com- higher than the other samples. pared to the other locations. The concentration of alkaline earth elements Ca, Sr and Ba The obtained data show that no serious artificial pollution was obtained for Ulva rigida, E.intestinalis and C. vagabunda The along our shore and element concentrations in algae during the data for Ulva and Enteromorpha showed no specific dependence studied period demonstrate considerably constant accumulation on location or time of sampling while the Cladofora values�� var- pattern. The studied green algae species are suitable for assessment ied to a greater extent. The average value for Sr in Ulva rigida is of element behavior in Black Sea marine ecosystems (especially lower (~150 µg/g) than those for Enteromorpha (~340 µg/g) and Ulva, Enteromorpha and Cladophora) and can be used as bio- Cladophora(~280 µg/g) with several samples with much higher indicators species for the ecological status of the ecosystem, values. The average value for the barium concentration for the together with the abiotic element concentration in the ecosystem three algae is close (~40 µg/g) also with few exceptions. The Ba (water, sediment). This work contributes to the evaluation of value in Tzarevo Sping`96 is exceptionally high 300 µg/g but the for heavy metal concentration in green Macroalgae and their sampling was performed close to the area of the port. We have ecological impact on marine ecosystems along the Bulgarian investigated several other green algae - Ulva lactuca, Bryopsis Black Sea coast. plumosa, Enteromorpha compressa,�� Cladophora coleotrix,�� Cha� etomorpha gracilis. The data obtained for Ulva lactuca species References were from locations Rusalka, Bjala, Tzarevo and Sizemore at the Bulgarian Black Sea coast. The determined element concentrations 1. Babich H, DevarisM, Stotzki G (1985) Mediation of Mutagenicity and Clastogenicity of Heavy Metals by Physiochemical Factors. Environ- are similar to those, obtained for Ulva rigida. The concentrations mental Research 37: 325-331. of the most of elements in Bjala location are higher compared to the other sites (like Ulva rigida) – for Fe – 1125 ± 50 µg/g, Cu – 14 2. Forsberg A, Soderlund S, Frank A, Petersson L, Pedersen M, et al. ± 2 µg/g, Zn – 78 ± 5 µg/g, Br - 807 ± 20 µg/g, Ba - 84 ± 5 µg/gSn (1988) Studies of metal concentration in the brown seaweed, Fucus vesiculosus from the Archipelago of Stockholm. Environmental Pol- - 28 ± 2 µg/g, Rb – 23 ± 2 µg/g. MN concentration is twice higher lution 49: 245-263. in two locations (Sinemorez Spring`98, Tzarevo Autumn`02) than the rest sites. Unlike Ulva rigida species, in Ulva lactuca Cd was 3. Rainbow P (1995) Biomonitoring of heavy metal availability in the marine environment. Marine Pollution Bulletin 31: 183-192. measured only in two locations - Tzarevo Autumn`02 - 4 ± 1 µg/g, Sinemorez Spring`05 - 6 ± 1 µg/g. 4. Rainbow P, Philips D (1993) Cosmopolitian biomonitors of trace metals. Marine Pollution Bulletin. 26 : 593-601. The data for Enteromorpha compressa are similar to those of E. intestinalis. The same is true for both the Cladophora algae 5. Favero N, Cattalini F, Bertaggia D, and Albergoni V (1996) Metal Ac- cumulation in a Biological Indicator (Ulva rigida) from the Lagoon of species (e.g. higher concentration of I). The measured values for Venice (Italy) Arch. Environmental Contamination Toxicology 31: 9 the different elements in Chaetomorphagracilis are closer to C. -18.

15 Volume 2018; Issue 01 Citation: Strezov A, Emilia N (2018) Study of Multielement Concentration in Bulgarian Black Sea Green Macroalgae. Curr Trends Oceanogr Mar Sci: CTOMS-103. DOI: 10.29011/ CTOMS-103.100003

6. Muse J, Stripeikis J, Fernandez F, Dhuicque L, Tudino M, Phillips D, 14. Topcuoglu S, Guven K, Kirbasoglu C, Gündör N, Unlu S, et al. (2001) et al. (1977) The use of biological indicator organisms to monitor trace Heavy metals in marine algae from Sile in the Black Sea, 1994-1997. metal pollution in the marine and estuarine environments - A review. Bulletin of Environmental Contamination Toxicology, 67: 288-294. Environmental Pollution 13: 281-317. 15. Topcuoğlu S, Esen N, Egilli E,Gündör N, Kut D, et al. (1998) Trace ele- 7. Ho Y (1990) Ulva lactuca as bio indicator of metal contamination in- ments and 137-Cs in Macroalgae and Mussels from the Kilyos in Black tertidal waters in Hong Kong. Hydrobiology 203: 73-81. Sea. International Symposium of Marine Pollution, Monaco, IAEA SM 354: 283-284. 8. Haritonidis S, Malea P (1995) Seasonal and local variation of Cr, Ni and Co concentration in Ulva rigida, C. agardh and Enteromorpha 16. Roeva N, Rovinski F, Kononov F(1996) Specific features of heavy linza (Linnaeus) from Thermaikos Gulf, Greece. Environmental Pollu- metal behavior in different nature media. Zhurnal Analytichnoy Khimii tion 89: 319-327. 51: 384 -391.

9. Haritonidis S, Malea P (1999) Bioaccumulation of metals by the green 17. Güven K, Topcuoglu S, Kut D, Esen N, Erentürk N, et al. (1992) Metal alga Ulva rigida from Thermaikos Gulf, Greece. Environmental Pol- uptake by Black Sea Algae. Botanica Marina 35: 337-340. lution 104: 365 - 372. 18. Phillips D (1977) The use of biological indicator organisms to monitor 10. Malea P, Haritonidis S (2000) Use of green alga Ulva rigida as an trace metal pollution in marine and estuarine environments - a review. indicator species to reassess metal pollution in the Thermaikos Gulf, Environmental Pollution, 13: 281-317. Greece, after 13 years. Journal of Applied Phycology 12: 169-176. 19. Burdin K, CruPina M, Saveljev I (1982) Black Sea Macroalgae as the 11. Locatelli C (1999) Sea water quality criteria by bioindicators. Pos- object for biogas-chemical monitoring of heavy metals. Chelovek i Bio- sible classification based on metal concentrations in Ulva rigida and sfera 7: 139 -145. Tapes philippinarum. Toxicological & Environmental Chemistry 71: 435-446. 20. Worldwide Proficiency Test for X-Ray Laboratories PTXRFIAEA/05 (2009) Determination of Minor & Trace Elements in Marine Sediment. 12. Strezov A (2008) Comparison of Some Radionuclides in the Marine IAEA Laboratories Seibersdorf. Coastal Environment of the Black Sea & the Mediterranean. J Eur- asian Analytical Chemistry 3: 170 -182. 21. Worldwide Proficiency Test for X-Ray Laboratories PTXRFIAEA/06 (2009) Determination of minor and trace elements in grass mixture. 13. Strezov A, NonovaTz (2009) Influence of Macroalgal Diversity on Ra- IAEA Laboratories, Seibersdorf. dionuclide and Heavy Metal Accumulation in Bulgarian Black Sea Eco- systems. J Environmental Radioactivity 100: 144 -150.

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