Bull Vet Ist Pulawy, 9-9, 201 DOI: 10.27/bvip-201-0091 Toxic elemets i free-livig freshwater fish, water a sedimets i Pola Józef Szkoda, Ja Żmudzki, Agieszka Nawrocka, Mirosława Kmiecik Departmet of Pharmacology a Toxicology Natioal Veteriary Research Istitute, 2-100 Pulawy, Pola szkoda@piwet.pulawy.pl Received: April 2, 201 Accepted: October, 201 Abstract Samples for aalysis were collected from 10 areas, icludig the major Polish rivers a lakes, with differet sources of evirometal pollutio (iustrial, muicipal, a farmig). The materials was take from the lakes of Mazury, located i a o-iustrialised regio, from the Brda River, a area impacted by pig farms, from the lakes of Lipczyo Wielkie/Pomeraia, from the Wkra River, a area impacted by poultry farms, from the Duajec River at the Rozowski Reservoir, from the Vistula River at Cracow a Warsaw, from the Odra River at Wroclaw a the Warta River estuary, a also from Rybik Power Statio Reservoir. Cocetratios of,,, a were aalysed i 397 fish muscle a 12 sedimet samples usig a atomic absorptio spectrometry techique. The aalytical procedures were covered by a quality assurace programme. It was demostrated that the average cocetratios of lead, cadmium, a arseic i fish were i the low huredths a thousaths of a mg/kg a ever exceeded permitted limits established for food. Higher values of these elemets were fou i fish from bodies of water located i the zoe of ifluece of large urba agglomeratios, especially the Cracow regio. High cocetratios of lead a cadmium were also fou i Vistula River sedimets ear Cracow, where the maximum values were 13.10 mg/kg a 21.2 mg/kg dry weight for lead a cadmium respectively. The average cocetratio of mercury i a predatory fish muscle (0.179 mg/kg) was almost twice as high as i the omivorous fish (0.103 mg/kg). Oly a sigle fish sample exceeded the maximum limit for this metal ( mg/kg) a did ot preset a risk to cosumers health. Keywords: rivers, lakes, fish, sedimets, toxic elemets, Pola. Itroductio The problem of presece of toxic elemets i fish a other aquatic orgaisms is coected with water pollutio as a result of atural processes a the release of hazardous compous to soil a air, as well as broader huma activity. Some elemets such as zic, iro, a copper, play a importat role i the biological system whe preset i sigificat amouts. I cotrast, lead, cadmium, mercury, or arseic ca be toxic, eve i trace amouts. The cocetratios of these elemets i fish a sedimets are the subject of scietific discussio i may publicatios (, 9, 10, 1, 22, 2, 2, 27-29). Lead a cadmium are widespread i the eviromet. For humas they pose a risk through cosumptio of cotamiated food a ihalatio (7, 12, 1, 29, 30). The predomiat form of mercury preset i the atmosphere is metallic mercury, which is almost 9% of the mercury emitted ito the eviromet. Biomethylatio plays a importat role i the movemet of mercury, where the products of this process ca easily trasfer amog differet compartmets of the eviromet. Both metallic mercury a its various compous cotaied i mieral deposits a sedimets are biomethylated (1). Methyl mercury compous are readily soluble i water a become very dagerous for fish a humas (). High bioaccumulative capacity for mercury i the trophic chai causes the cotamiatio of air, soil, water a food. Fish are a major source of mercury for livig orgaisms, especially for people, a they pose a relevat health risk (3, 13). I the case of arseic, iorgaic trivalet arseic compous are the most toxic to livig orgaisms. Uautheticated Dowload Date /17/1 1: PM
90 J. Szkoda et al./bull Vet Ist Pulawy/ (201) 9-9 Iorgaic arseic ca be methylated after risig up the food chai to humas or aimals, thus less toxic orgaic forms are created. The studies of biological samples idetified the occurrece of several tes of orgaic forms of arseic. Orgaic arseic compous such as arseobetaie (B) or arseocholie (C) are the domiat forms existig i aquatic orgaisms (algae, fish, crustaceas, a molluscs). May authors revealed that the orgaic arseic i fish, primarily i the form of arseobetaie, costitutes 7%-9% of the total arseic cotet (, 1, 17). The aim of the study was to evaluate the cocetratios of lead, cadmium, mercury, a arseic i selected species of free-livig freshwater fish ihabitig differet eviromets (rivers a lakes), a i water a sedimets from muicipal, iustrial, a farmig regios. Material a Methods Betwee 2011 a 2013, a aalysis was coucted of toxic elemets i differet species of freshwater fish, sedimets, a water obtaied from differet bodies of water. The samples were collected from 10 areas with differet sources of evirometal pollutio (iustrial, muicipal, a farmig). The material cosisted of omivorous a predatory fish (Table 1), which were take from locatios icludig Pola's major rivers a lakes (the lakes of Mazury, located i a o-iustrialised regio, the Brda River, a area impacted by pig farms, the Wkra River, a area impacted by poultry farms, the lakes of Lipczyo Wielkie/Pomeraia, the Duajec River at the Rozowski Reservoir, the Vistula River at Cracow a Warsaw, the Odra River at Wroclaw, the Warta River estuary, a Rybik Power Statio Reservoir). Table 1. List of examied fish species from rivers, lakes, a reservoirs 1 Samplig site Lake Łańskie Lake Maróz/Mazury 2 Wkra River/Żuromi 3 Brda River/Czluchów Lakes of Lipczyo Wielkie/Pomeraia Rozowski Reservoir Rybik Reservoir 7 Vistula/Cracow Vistula/Warsaw 9 Odra/Wroclaw 10 Odra/Warta estuary Omivorous fish roach (Rutilus rutilus) - *, bream (Abramis brama) - roach (Rutilus rutilus) 3, tech (Tica tica) - 1 roach (Rutilus rutilus) -, bream (Abramis brama) - 13, tech (Tica tica) - 9, ablet (Alburus alburus) - 2 roach (Rutilus rutilus) -, bream (Abramis brama) - roach (Rutilus rutilus) -, bream (Abramis brama) - roach (Rutilus rutilus) - 2, bream (Abramis brama) -, white bream (Blicca bjoerka) - roach (Rutilus rutilus) - 1, bream (Abramis brama) -, white bream (Abramis brama) - 9, barbell (Barbus barbus - 1 roach (Rutilus rutilus) -, bream (Abramis brama) - 1, tech (Tica tica) - 1 roach (Rutilus rutilus) - 9, bream (Abramis brama) -, white bream (Blicca bjoerka) - 1 roach (Rutilus rutilus) - 1, bream (Abramis brama) - 1, tech (Tica tica) - 2, white bream (Blicca bjoerka) - 2 Number of samples Predatory fish Number of samples 21 pike (Esox lucius) - 21* 21 37 29 12 pike (Esox lucius) -, perch (Perca fluviatilis) -, ide (Leuciscus idus) - 1 pike (Esox lucius) - 7, zaer (Saer lucioperca) - 10, perch (Perca fluviatilis) - 9, catfish (Silurus glais) - 3 pike (Esox lucius) -, zaer (Saer lucioperca) - 1 ot determied 0 2 3 23 zaer (Saer lucioperca) - 10, catfish (Silurus glais) -, perch (Perca fluviatilis) - 2 perch (Perca fluviatilis) -, chub (Squalius cephalus) -, catfish (Silurus glais) - 3 pike (Esox lucius) -, zaer (Saer lucioperca) -, asp (Leuciscus aspius) - 2, ide (Leuciscus idus) - 1, crucia carp (Carassius carassius) - 2, perch (Perca fluviatilis) - 2, catfish (Silurus glais) - 1 burbot (Lota lota) -, perch (Perca fluviatilis) - 1, catfish (Silurus glais) - 1, sumik (Ameiurus ebulosus) -, ide (Leuciscus idus) - 2, catfish (Silurus glais) - 1, asp (Leuciscus aspius) -1 pike (Esox lucius) -, perch (Perca fluviatilis) - 2, asp (Leuciscus aspius) - 2, ide (Leuciscus idus) - 1 Total 23 9 21 29 13 17 11 20 1 9 *- umber of fish aalysed Uautheticated Dowload Date /17/1 1: PM
J. Szkoda et al./bull Vet Ist Pulawy/ (201) 9-9 91 Cocetratios of lead, cadmium, mercury, a arseic were aalysed i 397 fish muscle samples (23 omivorous a 9 predatory) a 12 sedimet samples. Fish a sedimet samples were collected by the Veteriary Ispectorate a prepared i accordace with the istructios. After delivery to the laboratory, the fish were weighed, measured, a filleted. The, the samples of muscles were homogeised a stored i plastic bags at -20 C util aalysis. Sedimet a water samples were stored at C util aalysis. Determiatio of lead a cadmium i fish was performed by atomic absorptio spectroscopy with graphite furace atomisatio (PiAAcle 900T, Perki Elmer, USA), after dry digestio i a electric furace at 0 C. Arseic was determied by atomic absorptio spectrometry usig hydride geeratio techiques after digestio with magesium itrate i a electric furace at 0 C. Mercury aalysis was performed by atomic absorptio spectrometry usig a AMA 2 mercury aalyser (Altec, Czech Republic). For the digestio of sedimets pressure microwave digestio was used. The procedures for determiatio of lead, cadmium, arseic, a mercury i biological materials were previously described i details (2). Statistical calculatios were performed usig Statistica 10 PL (StatSoft, Pola). A importat part of the study was a special quality assurace programme for the aalytical methods used. The Natioal Referece Laboratory of the Departmet of Pharmacology a Toxicology participates i proficiecy tests (PT) orgaised by the Europea Uio Referece Laboratory (EURL- CEFAO, ISS, Rome, Italy) a also orgaises PT for regioal laboratories dealig with the aalysis of metals (20). The methods are also tested i the laboratory by aalysig certified referece materials (CRM) for trace elemets (Table 2). Results The results of aalysis of lead, cadmium, mercury, a arseic i the fish muscles are show i Table 3 a i Figs. 1 a 2. Lead. The average cotet of lead i the muscles of omivorous fish was 2 mg/kg. Twofold higher values were fou i fish take from the Rozowski Reservoir, the Vistula at Cracow a surrouig area, a the cofluece of the Odra a Warta Rivers (Fig. 1). The highest cocetratios of this elemet i predatory fish were fou i fish caught i the Odra River at the mouth of the Warta (Fig. 2). The average cocetratios of lead i sedimets raged from 2.0 to mg/kg dry weight. The cotet of lead i sedimet was high, with the maximum value of 13 mg/kg dry weight. It was fou i the sedimet from the Vistula River ear Warsaw (Table 3). Cadmium. The average cocetratio of cadmium (Figs. 1 a 2) i omivorous a predatory fish was low. It was at the level of mg/kg for the most regios. Oly i samples collected from the Vistula River ear Cracow the cadmium cocetratio was threefold higher at 09 mg/kg, with the maximum value beig for the roach at mg/kg (Table 3). I the sedimets, cocetratios of cadmium i secoary sludge raged from to.11 mg/kg. The highest cocetratio of this elemet, as it also was i the case of lead, was fou i the sedimet from the Vistula ear Cracow, where the maximum value reached 21.2 mg/kg dry weight (Table 3). Mercury. The average cocetratio of i the omivorous fish muscles was 0.103 mg/kg, while the highest value was fou i samples take from the Rozowski Reservoir, where the average cocetratio of this elemet reached 0.21 mg/kg (Fig. 1). I fish examied from other bodies of water, the average mercury cocetratio i muscle was at the low-teths a huredths of a mg/kg level (Figs. 1 a 2). The average cocetratio of mercury i the muscle of predatory fish was almost twofold higher tha that fou i omivorous fish a was 0.179 mg/kg. The lowest level of this elemet was fou i predatory fish caught i the Rybik Reservoir (Fig. 2). Table 3 summarises the results of the determiatio of mercury i the muscles of omivorous a predatory fish a i bottom sedimets. Noteworthy is the high cocetratio of mercury fou i the muscles of omivorous fish collected from the Vistula ear Warsaw a the Odra ear Wroclaw, where the average values of were 0.1 a 0.121 mg/kg respectively for roach a 0.11 a 0.11 mg/kg for bream, a were sigificatly higher tha those fou i fish of this species from the other examied bodies of water. I the aalysed muscles of predatory fish (pike, perch, a zaer), the highest average cocetratio of this elemet was fou i pike (0.292 mg/kg) a the maximum value i the sample from the Vistula River from Warsaw area was early double at 0.00 mg/kg (Table 3). Arseic. The average a maximum values of arseic (Fig. 1, Table 3) iicate that a high cocetratio of this elemet was i the muscles of omivorous fish. The highest cocetratio of this elemet was fou i fish caught i the Odra River ear Wroclaw. Arseic cocetratios fou i fish from this regio were much higher compared with regios outside large urba agglomeratios. I sedimet the mea cocetratios of arseic raged from 0.299 to.09 mg/kg dry weight (Table 3), while i water the cocetratio of the elemets was low a below the detectio limit of the method used ( mg/kg, mg/kg, mg/kg, a mg/kg). Uautheticated Dowload Date /17/1 1: PM
92 J. Szkoda et al./bull Vet Ist Pulawy/ (201) 9-9 Table 2. Cocetratios of elemets fou i proficiecy tests a certified referece materials Material Froze fish 1 th PT, EURL siged values (mg/kg) Z-scores 0.370 0.3 10-0.1 7 2 109 0. 0.222 0.211 9-0. 0.92 0.922 99. Froze meat 1 th PT, EURL siged values (mg/kg) Z-scores 1 9 9-0.2 100 097 100 103 0.3 - - - Froze fish PIWet-PT2012/ZFT/32 siged values Z-scores 09 0 9-0.37 100 0.11 0.11 100 102 7 DORM-3 Certified value (mg/kg) 0.39 0.37 93 0.290 0.27 9 0.32 0.3 9..29 91 mg/kg 0 0,0 0,1 0, 0,011 0,01 0,01 0,01 0,00 0,0 0,0 0,03 0,017 0,0 Lakes of Mazury Rybik Reservoir Brda River Lakes of Lipczyo W. Wkra River Rozowski Reservoir Vistula/Krakow Vistula/Warsaw Odra/Wroclaw Odra/estuary Warta 0,00 0,00 0,0 0,079 0,0 0,09 0,21 0,012 0,0 0,137 0,11 0,1 0,0 0,0 0,0 0,07 0,09 0,02 0,072 0,09 0,10 0,07 Fig. 1. Mea cocetratios of toxic elemets i muscles of omivorous fish mg/kg 0 0,0 0,1 0, 0,009 0,01 0,0 0,01 0,009 0,0 0,02 0,011 0,07 Lakes of Mazury Rybik Reservoir Brda River Lakes of Lipczyo W. Wkra River Rozowski Reservoir Vistula/Krakow Vistula/Warsaw Odra/Wroclaw Odra/estuary Warta 0,009 0,00 0,00 0,21 0,037 0,227 0,2 0, 0,092 0,27 0,103 0,29 0,029 0,0 0,037 0,03 0,03 0,0 0,032 0,11 0,031 Fig. 2. Mea cocetratios of toxic elemets i muscles of predatory fish Uautheticated Dowload Date /17/1 1: PM
J. Szkoda et al./bull Vet Ist Pulawy/ (201) 9-9 93 Table 3. The cocetratios of toxic elemets i the muscles of selected species of freshwater fish (mg/kg wet weight) a sedimets (mg/kg dry weight) Samplig site L. Łańskie L. Maróz/ Mazury Wkra River Żuromi Brda River Czluchów L. Lipczyo Wielkie Rozowski Reservoir Rybik Reservoir Vistula/ Cracow Vistula/ Warsaw Odra/ Wroclaw Odra/ Warta estuary - ot determied Elemets roach bream pike perch zaer sedimets mea max mea max mea max mea max mea max mea max 21 10 2 1 20 09 9 1.31 2.11 0 0.392 0.721 1 0.120 3 0.120 0.227 0.30 37 2 9 0.100 22 29 29 2 3.77.13 3 9 9 13 7 3 92 27 1 9 11 7 0.10 3 0.1 0.13 9 10 0.121 0.117 11 0.22 0.111 1 0.11 73 13 0.11 0.122 0.101 0.32 0 0.239 0.222 0.22 0.20 17 27 1 0 1 0 0 1 3 0.11 0.11 1 0 0.133 0 0.391 9 0.,002 32 7 7 7 7 72 10 0.102 1 09 0.22 0.111 0.29 0.129 0.1 0.309 0.29 10 0. 13 0.27 21 13 0.27 21 32 0.30 0.1 0.3 0.3 0 0.17 20 7 0.13 2 0.27 0.291 33 0 0.292 0.00 2 3 9 3 0.1 29 0.233 32 1 1 0.20 0.27 10 7 1 37 2 0. 17 07 0.137 0.19 29 07 0. 29 0.322 22 22 0.207 2 11 0.21 0.30 10 2.09 0.9 0.3 2.2 0.299 10 1.33 1 0.10. 0.33.09 2. 0.139 0.1 3.99.11 0.9.01 21 2. 0.312 2.03 20 7.3 0.32 3. 2 7.9 0. 33 1.0 2.910 0.1 1..2 2 0.00 2.020 0.27 1. 0. 1.9.21 0.23 09 0.73 13.1 21.237 2.779 1. 13.1 0.921 0.1 10.2 20.3 1.02 0.12 1.27 0.709 3 1. Discussio Acceptable levels of lead, cadmium, a mercury i freshwater fish are set out i Europea Commissio Regulatio (EC) No. 11/200, a are as follows: 0.30 mg/kg, mg/kg, a mg/kg (). For arseic however, at Europea Uio maximum permissible level has ot bee established. It is still uer discussio a it is expected that the eed will emerge for the dual determiatio of the presece of this elemet, both i total a separately i orgaic a iorgaic forms. I view of the high toxicity of arseic, may EU Member States have set atioal actio levels for this elemet i fish. I Pola, there are actio levels set out i the "Natioal Residue Cotrol Pla for the iterpretatio of test results of arseic i foods of aimal origi, a the Pla is aually approved by the Chief Veteriary Officer a Europea Commissio (). A maximum permissible total arseic cotet i the muscles of fish of mg/kg has bee adopted i Pola. The preset study demostrated that the average cocetratios of lead, cadmium, a arseic i fish were i the low huredths a thousaths of a mg/kg a did ot exceed permitted limits i ay samples (3, ). Similar values of lead a cadmium i muscles of Uautheticated Dowload Date /17/1 1: PM
9 J. Szkoda et al./bull Vet Ist Pulawy/ (201) 9-9 river freshwater fish were detected by other authors (7, 12, 1, 23). By cotrast, higher cocetratios of lead, ofte exceedig 1 mg/kg, were fou i lake fish (, 1). I this study, higher values of these elemets were fou i fish from bodies of water located i the zoe of ifluece of large urba agglomeratios. Cocetratios of these elemets detected i fish from the Vistula ear Cracow, where the observed values were higher compared to other regios, are particularly importat. The average cocetratio of mercury i the muscles of predatory fish (0.179 mg/kg) was sigificatly higher tha that fou i omivorous fish (0.103 mg/kg). The lowest level of this elemet was fou i predatory fish caught i Rybik Reservoir (Fig. 2). I iividual fish samples (asp 0.33 mg/kg, a ide 0. mg/kg), cocetratios exceeded the acceptable level, i.e. 0. mg/kg. I fish breedig, mercury was fou at the level of mg/kg (2, 2, 27). I studies of differet species of fish caught i the river Vistula, mercury was fou i similar cocetratios (2). I the Czech Republic, this elemet was preset i fish i cocetratios ragig from to 0.32 mg/kg (10). The arseic cocetratio i the muscles of freshwater fish determied i this study seems ot to be a major problem, regardless of its place of origi. Both the average a maximum arseic cotets i fish i all the regios studied were low a did ot exceed the approved limit, i.e. mg/kg (). Higher levels of this elemet were fou i omivorous fish, particularly i roach, where the average for this elemet ( mg/kg) was threefold higher tha that fou i perch (2 mg/kg). I the study coucted i Frace, arseic i freshwater fish was fou i cocetratios ragig from 2 to 0.7 mg/kg (), a i studies coucted i Italy, the results raged from 0 to 10 mg/kg (23). Higher values of the aalysed elemets were fou i the sedimets, which were collected from the surface layer to a depth of 20 cm. Evaluated agaist the criteria for bottom sedimets, the level of lead i several samples exceeded the geochemical backgrou level, which is mg/kg dry weight. For other elemets studied, i.e. cadmium, mercury, a arseic, geochemical backgrou values are as follows: 0. mg/kg dry weight, < mg/kg dry weight, a < mg/kg dry weight. (2, 11). The highest cocetratios of lead, cadmium, a mercury were fou i sedimets collected from the Vistula ear Cracow. I all tested samples from this regio the cocetratios of these elemets exceeded the geochemical backgrou value. I additio, a large umber of samples i this regio exceeded the probable effects levels (PEL), which specify the cocetratio of cotamiatig elemets i sedimet, over which harmful effects o aquatic orgaisms are ofte observed (2, ). The observed cocetratios of mercury i sedimets i this regio (mea 0.9 a maximum 2.779 mg/kg dry matter) had o great effect o the bioavailability of mercury i fish, because its cocetratio i the examied fish (Table 3) was sigificatly lower compared with other regios a lower tha the allowable limit (). I the sedimets from other rivers a lakes i Pola geochemical backgrou values were exceeded oly i a small percetage of samples. The PEL was exceeded oly for lead (91 mg/kg dry matter) i three sedimet samples collected from the Vistula River i Warsaw. The studies coucted by other authors i Pola (9, 21, 2), obtaied values exceedig the geochemical backgrou a PEL. I river sedimets, the mea values of arseic were lower tha the permissible backgrou ( mg/kg), but the maximum value of arseic (1. mg/kg) i the sedimet of the Vistula ear Cracow iicates a higher arseic cotamiatio i compariso with other regios. Other authors fou that arseic cocetratio i sedimets of the Odra raged from.39 to 2 mg/kg (9). The study iicated that the cocetratios of toxic elemets i free-livig freshwater fish are rather low a safe. The mea values of lead a cadmium i omivorous a predatory fish did ot differ sigificatly amog them. However, i the case of mercury, the average cocetratio of this elemet i predatory fish muscles was almost twofold higher i compariso with omivorous fish. The studies have also show that oly a sigle fish sample exceeded the maximum limit for this metal ( mg/kg) (). I studies o the arseic cocetratio, higher values of this elemet were fou i the muscles of omivorous fish. The highest value of this elemet was fou i fish caught i the Odra a Warta River estuary; however, they do ot pose a risk to cosumers health. High cocetratios of lead, cadmium, mercury, a arseic were fou i sedimet samples collected from the Vistula ear Cracow, where a large umber of samples exceeded the value of geochemical backgrou a PEL for these elemets. 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