Jelena Stanković, Maja Raković, Momir Paunović, Ana Atanacković, Jelena Tomović, Đurađ Milošević

DOI Number
First page
Last page


The study was conducted on the Danube River, within the project Joint Danube Survey 3 (JDS3). The main aim was to estimate the quantity of microplastics in aquatic ecosystems through passive biological monitoring. Three freshwater species were used for microplastic (MP) isolation from different taxonomic groups of organisms: Mollusca, Oligochaeta, and Chironomidae (Diptera), with the following species: Lithoglyphus naticoides (C. Pfeiffer, 1828), Limnodrilus hoffmeisteri (Claparede, 1862), and Chironomus acutiventris (Wülker, Ryser & Scholl, 1983), respectively. The samples were collected from 6 sites along the Danube River where 540 specimens were examined. The samples were digested by alkaline method (incubation in 10% KOH solution at 60 ⁰C for 24 h) and filtered through a mill silk, 10 µm mesh size. Collected particles were categorized as: fibre, hard plastic, nylon, rubber, or miscellaneous. Categories were divided into subcategories based on the coloration of the particles. Particles ingested by organisms were represented mostly by fibres and fragmented hard plastics, within the size range were from 0.03 to 4.87 mm. A total of 678 MP particles were collected with an average of 4.64 ± 1.59; 1.64 ± 0.46 and 1.24 ± 0.34 items/organism isolated from L. hoffmeisteri, L. naticoides and C. acutiventris, respectively. According to results, L. hoffmeisteri, L. naticoides and C. acutiventris could be used as proper bioaccumulators of MP pollution in the Danube River.


microplasic, plastic litter, freshwater, alkaline method, macrozoobenthos, the Danube River

Full Text:



Plastics Europe. Plastics — the facts 2020. An analysis of European plastics production, demand and waste data. 2020.

Thompson RC, Swan SH, Moore CJ, vom Saal FS. Our plastic age. Philos Trans R Soc Lond B Biol Sci 2009; 364:1973–1976.

Gigault J, ter Halle A, Baudrimont M, et al. Current opinion: What is a nanoplastic? Environ Pollut 2018; 235:1030-1034.

Fendall LS, Sewell MA. Contributing to marine pollution by washing your face: MPs in facial cleansers. Mar Pollut Bull 2009; 58:1225–1228.

Lassen C, Hansen SF, Magnusson K, et al. Microplastics - Occurrence, effects and sources of releases to the environment in Denmark. The Danish Environmental Protection Agency 2015.

Sherrington C, Darrah C, Hann S, Cole G, Corbin M. Study to support the development of measures to combat a range of marine litter sources, Eunomia Research & Consulting Ltd, Bristol, United Kingdom 2016.

Andrady. Microplastics in the marine environment. Mar Pollut Bull 2011; 62:1596–1605.

Endo S, Takizawa R, Okuda K, et al. Concentration of polychlorinated biphenyls (PCBs) in beaches resin pellets: variability among individual particles and regional differences. Mar Pollut Bull 2005; 50:1103–1114.

Ogata Y, Takada H, Mizukawa K, et al. International Pellet Watch: global monitoring of persistent organic pollutants (POPs) in coastal water.1. Initial phase on PCB, DDT and HCHs. Mar Pollut Bull 2009; 58:1437–1446.

Hirai H, Takada H, Ogata Y, et al. Organic micropollutants in marine plastics debris from the open ocean and remote and urban beaches. Mar Pollut Bull 2011; 62:1683–1692.

Oehlmann JR, Schulte-Oehlmann U, Kloas W, et al. A critical analysis of the biological impacts of plasticizers on wildlife. Philos Trans R Soc B 2009; 364:2047−2062.

Barnes DKA, Galgani F, Thompson RC, Barlaz M. Accumulation and fragmentation of plastic debris in global environments. R Soc 2009; 364:1985-1998.

Dris R, Gasperi J, Rocher V, Saad M, Renault N, Tassin B. Microplastic contamination in an urban area: a case study in Greater Paris. Environ Chem 2015; 12:592–599.

Peters CA, Bratton SP. Urbanization is a major influence on microplastic ingestion by sunfish in the Brazos River Basin, Central Texas, USA. Environ Pollut 2016; 210:380–387.

Lechner A, Keckeis H, Lumesberger-Loisl F, et al. The Danube so colourful: a potpourri of plastic litter outnumbers fish larvae in Europe’s second largest river. Environ Pollut 2014; 188:177–181.

Morritt D, Stefanoudis PV, Pearce D, Crimmen A, Clark P.F. Plastic in the Thames: a river runs through it. Mar Pollut Bull 2014; 78:196–200.

Su L, Xue Y, Li L, et al. Microplastics in Taihu Lake, China. Environ Pollut 2016; 216:711–719. 10.1016/j.envpol.2016.06.036

Eriksen M, Mason S, Wilson S, et al. 2013. Microplastic pollution in the surface waters of the Laurentian Great Lakes. Mar Pollut Bull 2013; 77:177–182.

Free CM, Jensen OP, Mason SA. High levels of microplastic pollution in a large, remote, mountain lake. Mar Pollut Bull 2014; 85:156–163.

Corcoran PL, Norris T, Ceccanese T,Walzak MJ, Helm PA, Marvin CH. Hidden plastics of Lake Ontario, Canada and their potential preservation in the sediment record. Environ Pollut 2015; 204:17–25.

Biginagwa F, Mayoma B, Shashoua Y, Syberg K, Khan F. First evidence of microplastics in the African Great Lakes: recovery from Lake Victoria Nile perch and Nile tilapia. J Great Lakes Res 2016; 42:1146–1149.

Imhof HK, Ivleva NP, Schmid J, Niessner R, Laforsch C. Contamination of beach sediments of a subalpine lake with microplastic particles. Curr Biol 2013; 23:867–868.

Klein S, Worch E, Knepper TP. (2015). Occurrence and spatial distribution of MPs in river shore sediments of the Rhine-Main area in Germany. Environ Sci Technol 2015; 49:6070–6076. est.5b00492

Zbyszewski M, Corcoran PL. Distribution and Degradation of Fresh Water Plastic Particles Along the Beaches of Lake Huron, Canada. Water Air Soil Pollut 2011; 220:365–372.

Firdaus M, Trihadiningrum Y, Lestari P. Microplastic pollution in the sediment of Jagir Estuary, Surabaya City, Indonesia. Mar Pollut Bull 2019; 150:110790.

Rech S, Macaya-Caquilpán V, Pantoja JF, Rivadeneira MM, Madariaga DJ, Thiel M. Rivers as a source of marine litter – A study from the SE Pacific. Mar Pollut Bull 2014; 82: 66–75.

Yonkos LT, Friedel EA, Perez-Reyes AC, Ghosal S, Arthur CD, 2014. Microplastics in four estuarine rivers in the Chesapeake bay U.S.A. Env Sci Tech 2014; 48:14195–14202.

Zhao S, Zhu L, Wang T, Li D. Suspended microplastics in the surface water of the Yangtze Estuary System, China: First observation on occurrence, distribution. Mar Pollut Bull 2014; 86:562–568.

Jovanović B, 2017. Ingestion of microplastics by fish and its potential consequences from a physical perspective. Integrated Environ Assess Manag 2017; 13: 510–515.

Gregory MR. Environmental implications of plastic debris in marine settings—entanglement, ingestion, smothering, hangers-on, hitch-hiking and alien invasions. Philos Trans R Soc B 2009; 364:2013–2025.

Zettler ER, Mincer TJ, Amaral-Zettler LA. Life in the “Plastisphere”: Microbial Communities on Plastic Marine Debris. Env Sci Tech 2013; 47:7137–7146.

Teuten EL, Saquing JM, Knappe DRU, et al. Transport and release of chemicals from plastics to the environment and to wildlife. Philos Trans R Soc Lond B Biol Sci 2009; 364: 2027–2045.

Masó M, Garcés E, Pagès F, Camp J. Drifting plastic debris as a potential vector for dispersing Harmful Algal Bloom (HAB) species. SCI MAR 2003; 67:107–111.

McCormick A, Hoellein T, Mason SA, Schluep J, Kelly JJ. Microplastic is an abundant and distinct microbial habitat in an urban river. Env Sci Tech 2014; 48:11863–11871.

EU (2000) Directive 2000/60/EC of the European Parliament and of the Council of 23 October 2000 establishing a framework for Community action in the field of water policy. Water Framework Directive (WFD).

European Commission (2014) COM/2014/0397. Proposal for a Directive of the European Parliament and of the Council amending Directives 2008/98/EC on waste, 94/62/EC on packaging and packaging waste, 1999/31/EC on the landfill of waste, 2000/53/EC on endof- life vehicles, 2006/66/EC on batteries and accumulators and waste batteries and accumulators, and 2012/19/EU on waste electrical and electronic equipment.

European Commission (2013) GREEN PAPER: On a European Strategy on Plastic Waste in the Environment.

European Commission (2015) Closing the loop: Commission adopts ambitious new Circular Economy Package to boost competitiveness, create jobs and generate sustainable growth. Press release, Brussels, 2 December 2015, IP/15/6203.

EU (2006) Regulation (EC) No 1907/2006 of the European Parliament and of the Council of 18 December 2006 concerning the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), establishing a European Chemicals Agency, amending Directive 1999/ 45/EC and repealing Council Regulation (EEC) No 793/93 and Commission Regulation (EC) No 1488/94 as well as Council Directive 76/769/EEC and Commission Directives 91/155/EEC, 93/67/EEC, 93/105/EC and 2000/21/EC.

Carpenter EJ, Smith KL. Plastics on the Sargasso Sea Surface. Science 1972; 175:1240–1241.

Black Sea Commision. Strategic Action Plan for the Environmental Protection and Rehabilitation of the Black Sea. 2009.

Sommerwerk N, Schneider-Jajoby M, Baumgartner C, et al. The Danube River Basin. In: Tockner K, Robinson CT, Uehlinger U (eds) Rivers of Europe. Elsevier Ltd., London, UK, 2009. 59-113.

ICPDR - International Commission for the Protection of the Danube River. Joint Danube Survey 3- Full report on: Macroinvertebrates. 2014. (

Atanacković AD, Šporka F, Csányi B, Vasiljević BM, Tomović JM, Paunović MM. Oligochaeta of the Danube River – a faunistical review. Biol 2013; 68:269–277.

Paunović MM, Csányi B, Simonović P, Zorić K, 2015. Invasive Alien Species in the Danube. In: Liska I (eds) The Danube River Basin. The Handbook of Environmental Chemistry, vol 39. Springer, Berlin, Heidelberg 2015.

Pinder L. Biology of freshwater chironomidae. Annu Rev Entomol 1986; 31:1-23.

Scherer C, Brennholt N, Reifferscheid G, Wagner M. Feeding type and development drive the ingestion of microplastics by freshwater invertebrates. Sci Rep 2017; 7:17006.

AQEM. Manual for the application of the AQEM system. A comprehensive method to assess European streams using benthic macroinvertebrates, developed for the purpose of the Water Framework Directive 2002; Contract No:EVK1-CT1999-00027).

Liška I, Wagner F, Sengl M, Deutsch K, Slobodník J. Joint Danube Survey 3: A Comprehensive Analysis of Danube Water Quality. International Commission for the Protection of the Danube River and European Commission, Vienna 2015.

Moller Pillot H. De larven der Nederlandse Chironomiae (Diptera). 1A: Inleiding, Tanypodinae en Chironomini. St E I S Nederland, Leiden (German) 1984a.

Moller Pillot, H. De larven der Nederlandse Chironomiae (Diptera). 1B: Orthocladiinae sensu lato. St E I S Nederland, Leiden (German) 1984b.

Schmid P. A Key to the Larval Chironomidae and their instars from Aus-trian Danube Region Streams and Rivers: Part 1. Diamesinae, Prodiamesinae and Orthocladiinae. Federal Institute for Water Quality of the Ministry of Agricultureand Forestry, Wien 1993.

Vallenduuk HJ, Moller Pillot H. Chironomidae Larvae of the Netherlands and Adjacent Lowlands: General Ecology and Tanypodinae. KNNV Publishing, Zeist 2007.

Pfleger V. A field guide in colour to Molluscs. Silverdale Books, Prague, Czech 2000:216.

Timm T. A quide to the freshwater Oligochaeta and Polychaeta of Northern and Central Europe. Lauterbornia 2009; 66:1–235.

Claessens M, Van Cauwenberghe L, Vandegehuchte MB, Janssen CR. New techniques for the detection of microplastics in sediments and field collected organisms. Mar Pollut Bull 2013; 70:227–233.

De Witte B, Devriese L, Bekaert K, et al. Quality assessment of the blue mussel (Mytilus edulis): Comparison between commercial and wild types. Mar Pollut Bull 2014; 85, 146–155.

Cole M, Webb H, Lindeque PK, Fileman ES, Halsband C, Galloway TS. Isolation of microplastics in biota-rich seawater samples and marine organisms. Sci Rep 2014; 4:4528.

Foekema EM, De Gruijter C, Mergia MT, van Franeker JA, Murk AJ, Koelmans AA. Plastic in north sea fish. Env Sci Technol 2013; 47:8818–8824.

Nuelle MT, Dekiff JH, Remy D, Fries E. A new analytical approach for monitoring microplastics in marine sediments. Environ. Pollut 2014; 184:161–169.

Avio CG, Gorbi S, Milan M, et al. Pollutants bioavailability and toxicological risk from microplastics to marine mussels. Environ Pollut 2015; 198:211–222.

Dehaut A, Cassone AL, Frère L, et al. Microplastics in seafood: Benchmark protocol for their extraction and characterization. Environ. Pollut 2016; 215:223–233.

Ferreira T, Rasband W. ImageJ User Guide. 2012.


Woodall LC, Sanches-Vidal A, Canals M, et al. The deep sea is a major sink for microplastic debris. R Soc Open Sci 2014; 1: 140317.

Mani T, Hauk A, Walter U, Burkhardt-Holm P. Microplastics profile along the Rhine River. Sci Rep 2015; 5:1–7.

Horton AA, Jürgens MD, Lahive E, van Bodegom PM, Vijver MG. The influence of exposure and physiology on microplastic ingestion by the freshwater fish Rutilus rutilus (roach) in the River Thames, UK. Environ Pollut 2018; 236:188–194.

Xu Q, Deng T, LeBlanc GA, An L. An effective method for evaluation of microplastic contaminant in gastropod from Taihu Lake, China. Environ Sci Pollut Res 2020; 27:22878–22887.

Akindele EO, Ehlers SM, Koop JHE. First empirical study of freshwater microplastics in West Africa using gastropods from Nigeria as bioindicators. Limnologica 2019; 78:125708.

Hurley RR, Woodward JC, Rothwell JJ. Ingestion of Microplastics by Freshwater Tubifex Worms. Environ Sci Technol 2017; 51:12844–12851.

Lin CT, Chiu MC, Kuo MH. Effects of anthropogenic activities on microplastics in deposit-feeders (Diptera: Chironomidae) in an urban river of Taiwan. Sci Rep 2021; 11:400.


Hohenblum P, Liebmann B, Liedermann M. Plastic and Microplastic in the environment. The Environment Agency Austria, Vienna 2015.

Stanković J, Milošević Dj, Paunović M, et al. Microplastic ingestion by freshwater Asian clams Corbicula fluminea (Müller, 1774) in the Danube River and main tributaries. Manuscript in



  • There are currently no refbacks.

© University of Niš, Serbia
Creative Commons licence CC BY-NC-ND
ISSN 0354-4699 (Print)
ISSN 2406-050X (Online)