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Supplement 1 Ecological Engineering and Environment Protection, IX, 2017, p.32-38

ECOLOGICAL STATUS OF UPPER PART OF RIVER CATCHMENT ACCORDING REGULATION N4 BASED ON MACROZOOBENTHOS AND FISH FAUNA Lyubomir Kenderov, Dimitriy Dashinov, Emil Kanev, Lyubomira Lyubomirova, Eliza Uzunova

Abstract: Recent information about the ecological status of upper part of Iskar River and some tributaries was given. Ecological assessment according WFD and national water legalisation (Regulation N4) was applied. For estimation of fish based indices (BRI and TRI) the following ichthyologic parameters were used: total abundance and biomass of ichthyofauna, abundance and biomass of Salmo trutta, occurrence of introduced species, sensitive and migrants. For calculation of BI and TTN indices were used taxa composition of the macroinvertebrate fauna. Ecological situation of Iskar River and tributaries was assessed as “high” according macroinvertebrate based indices and “moderate” to “bad” according fish based indices. Deterioration of ecological quality of lower river sites was recorded. Keywords: ecological status, fishes, macrozoobenthos, Iskar River

INTRODUCTION Reservoir) and on the other hand, they are valuable places for preserving biodiversity. In recent years Iskar River is the longest Bulgarian river (368 there has been a growing tourist activity and km) and flows through highly urbanized regions construction of tourist complexes and local ( City) and agriculture territories ( household pollution. Valley, , Western Valley). The The aim of this study is to give recent ecological status and water quality of Iskar River in information about ecological situation of mountain its middle and lower reaches during the 70s to 90s of part of Iskar River catchment according different the 20th century has been very poor [6, 16,17, 19]. biotic indices using fish and macrozoobenthos This is caused by the household water loading, communities. industrial pollution (including heavy metals) and loading of nitrates and phosphates from agriculture MATERIAL AND METHODS [12]. The latest studies (2004 - 2006) show a significant improvement in the ecological situation Sampling sites [8]. The worst pollution (bad ecological status or For the purposes of this study we selected 17 polysaprobity) is limited only in the area sampling stations along the upper Iskar River and its downstream Sofia region. Along the lower part of tributaries (Fig.1). Sampling sites were selected the river, significant self-purification processes are according potential anthropogenic impact from realized (up to β-mesosaprobity). On the contrary, in villages and hydroengendering structures. The site the mountainous part of the Iskar River ( codes and their geographical coordinates are Mountain), the ecological situation has always been presented in Table 1. acceptable [9, 16]. The main tributaries that form the Iskar River are the Beli Iskar, Cherni Iskar, Levi Fish sampling Iskar, Maliovishka, Preka, Lopushka, Pravi Iskar, Fish sampling was conducted during the low Lakatitsa, Musalenska Bistritza, as well as numerous water level period (July-August-September, 2015- smaller streams. The upper reaches of these rivers 2016). For assessment of fish abundance run through the territory of “Rila” National Park. electrofishing has been applied following the CEN The waters of some of them are included in a standard [1]. A single electrofishing transect drinking water supply system. In the lower reaches without block nets was applied in every testing of these rivers are situated small villages, several site. Sampling transects were between 50 to 165 m HPPs and numerous water abstractions. long, depending on river width (which varied from Consequently, the main negative factors that are 5 to 30 m). The total fishing area per site varied expected to influence the ecological status in these between 380 to 1100 m2. The electrofishing was mountain rivers are fragmentation and organic water conducted from one bank to the other over the pollution. The control of the ecological status of entire river stretch. The collected fish were these river stretches is important because they have counted and body mass was measured (total the meaning of drinking water (flow into the Iskar

32 Supplement 1 Ecological Engineering and Environment Protection, IX, 2017, p.32-38 weight, W in 0.1 g) in situ and then returned to the samples were collected. Sampling was performed locations of their capture. during June 2015 – August 2016. The taxonomic status of the species was Data analyses determined in situ according to [10]. The conservation status was determined according to the The fish abundance was expressed as number of Biological Diversity Act (OJ 77/2002), EU fishes caught per 1 ha and the biomass in kg per 1 legislation and international conventions. ha. For sites located in river type R4 index BRI [11] was used. For sites located in river types R1 and R2 Macrozoobenthos sampling Trout River Index, TRI [15] was used. Macroinvertebrates were sampled using hand net Ecological assessment according macrozoobenthic or Surber bottom sampler [4]. Up to 10 sub-samples community was done by metrics “Biotic index” (BI) and in one site using the “multihabitat approach” [5] were “Total taxa number” (TTN), Regulation N4 (OJ collected. Sampling of the predominant substrate - 22/2013). Metric ranges for river types “R2” (mountain stones and gravel from up to 100m river sector in rivers) “R4” (semi-mountain rivers) after national river each site was preferred. Totally 19 macrozoobenthic typology [2] were used.

Fig. 1. Site map of upper part of Iskar River catchment. (Site codes are explained in Table 1)

Table 1. Sampling sites of Iskar River catchment during 2015 - 2016 Site Altitude Sites (river, place) Geographic coordinates codes (m) N42.289183° Iskar River upstream Samokov Town IR.S 1035 E23.535658° N42.363972° Iskar River near Dragushinovo Village IR.D 895 E23.555447° N42.393161° Iskar River near Zlokuchene Village * IR.Z 865 E23.552722° N42.213806° Beli Iskar River upstream Beli Iskar Village BI.B 1420 E23.548083° N42.254814° Cherni Iskar River upstream Village CI.G 1180 E23.451667° N42.265933° Lakatitsa River upstream Govedartsi Village LA.G 1140 E23.469900° N42.243531° Levi Iskar River upstream Mala Tsarkva Village LI.U 1295 E23.516719° N42.270510° Levi Iskar River near Mala Tsarkva Village LI.M 1130 E23.508550°

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N42.225389° Lopushka River 3 км upstream Govedartsi LO.G 1580 E23.433419° River downstream Shiroki Dol N42.398111° PA.S 855 Village* E23.526722° Pravi Iskar River 4km upstream inflow to Cherni N42.215050° PI.C 1850 Iskar E23.341789° Preka River 1km upstream inflow to Cherni Iskar N42.229950° PR.C 1385 River E23.406261° N42.186850° Urdina River downstream Urdin Circus UR.U 1955 E23.345219° Beli Iskar River downstream Beli Iskar Village N42.285611° BI.Q 1058 ** E23.534992° Levi Iskar River downstream Mala Tsarkva N42.270958° LI.K 1125 Village ** E23.507312° N42.232912° Maliovishka River ** MA.L 1374 E23.392160° N42.319980° Musalenska Bistritza upstream Samokov town ** MU.B 979 E23.560094° * Only macroinvertebrates was investigated ** Only fish fauna was investigated

RESULTS AND DISCUSSION (Fig. 2). The main reason for the observed scores is the low carrying capacity of the high-mountain Fish study rivers in relation to the fish fauna. The numerous A total of 9 fish species belonging to 5 river fragmentations along studied rivers have families were caught in the investigated zone. additional negative impact on the size Three of the species are listed in Annex II of the composition of the ichthyofauna [7, 18]. In the European Commission Council Directive areas around villages, fishing (legal and illegal) 92/43/EEC on the conservation of natural habitats can also affect the fish abundance and biomass. and wild fauna and flora (Habitats Directive): However, due to the scarce scientific information Barbus petenyi, Sabanejewia balcanica and about the fish fauna in the past, this cannot be Cottus gobio. Some of the species are European claimed with certainty [13, 14]. General lower endemits [10] (Table 2). The brown trout Salmo values of fish indices observed in the upper Iskar trutta was caught at all investigated for fish sites, River are probably associated with the specifics in except one (IR.D). According to the ecological the hydrological regime of rivers in the region. It characteristics, the fish communities were is characterised by periods of extremely high dominated by rheophilic species (82 %). The water levels and velocities during the snow melt environmental status of the surveyed sites and rain in the spring, followed by extremely low according to the used fish indices belongs to the water levels in some rivers during the summer- categories “moderate”, “good”, “bad” and “very autumn period. Unsustainable hydrological bad” (Table 3). The stations located above conditions over the years require conducting settlements were assessed mainly in category more frequent monitoring studies [3]. “bad” and stations below - in category “moderate”

Table. 2. Fish species presented in the sampling sites in upper reaches of the Iskar River during the period 2015 – 2016

Fish species Sampling sites IR.D IR.S CI.G BI.Q BI.B LI.K LI.M LA.G LO.G MA.L PI.C PR.C UR.U MU.B Barbus petenyi + + + + Salmo trutta + + + + + + + + + + + + + + Gobio gobio + + + + Phoxinus phoxinus + + + + + + + Sabanejewia + + + balcanica

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Barbatula + + barbatula Cobitis strumicae + Cottus gobio + + + + +

Tab. 3. Ecological assessment of the sampling sites in upper reaches of the Iskar River, based on fish fauna indices (BRI and TRI), 2015-2016 Sampling sites Ecological status IR.D IR.S CI.G BI.Q LI.K LI.M LA.G LO.G MA.L PR.C PR.C UR.U MU.B very according good good good bad bad moderate bad good moderate bad moderate moderate* bad fish fauna * * ** ** ** ** ** ** ** ** ** ** * calculated according BRI ** calculating according TRI

Fig. 2. Distribution of environmental assessments by fish indices (BRI, TRI) in the river areas situated downstream the villages (left) and upstream the villages (right) in upper River Iskar and tributaries, 2015-2016

Macrozoobenthos study communities: dominant taxa were relatively tolerant to organic pollution (Erpobdella octoculata Linnaeus, Ecological status according the biological 1758 – Hirudinea; Tubificidae g.sp. – Oligochaeta; quality element “macrozoobenthos” was “high” in Hydropsyche incognita Pitsch, 1993 – Trichoptera; most of the cases (Table 4). Biotic index has maximal Chironomidae g.sp. – Diptera;). Taxa with declining score values (BI = 4/5 or 5) with the exception of abundance were xenosaprobic or oligosaprobic PA.S and IR.Z sites. In the last cases both rivers have indicators (Gammarus balcanicus Schaferna, 1922 – “good” ecological status according the BI (BI=4) due Amphipoda; Epeorus sp., Ecdyonurus sp., to deterioration of ecological quality. Such situation Rithrogena sp. – Ephemeroptera; Protonemoura sp., demonstrated TTN index– most sites have relatively Perla marginata Panzer, 1799, Perlodes intricatus big number of taxa, except lower parts of the Pictet, 1841 – Plecoptera, Blepharicera fasciata, catchment (IR.D, IR.Z and PA.S). In these sites we Liponeura sp. – Blephariceridae, Diptera). found some changes of macrozoobenthos

Table 4. Ecological quality (EQ) of upper part of Iskar River catchment after BI and TTN indices (Regulation N4) during 2015 – 2016

Site codes Date BI EQ TTN Site codes Date BI EQ TTN IR.S 2.6.2015 5 high 33 LA.G 19.8.2015 5 high 27 IR.S 11.8.2015 5 high 20 LI.U 21.8.2015 5 high 20 IR.D 2.6.2015 5 high 29 LI.M 12.8.2016 5 high 30 IR.D 11.8.2015 4,5 high 21 LO.G 30.7.2016 5 high 23 IR.Z 18.8.2015 4 good 21 PA.S 19.8.2015 4 good 20 BI.B 2.6.2015 5 high 27 PA.S 11.7.2016 4 good 19 BI.B 20.8.2015 5 high 23 PI.C 4.8.2016 5 high 19 CI.G 2.6.2015 5 high 36 PR.C 11.8.2016 5 high 26 CI.G 7.8.2015 4,5 high 23 UR.U 10.8.2016 5 high 25 CI.G 9.6.2015 5 high 21

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CONCLUSION and devices for benthic macroinvertebrates in fresh waters. In summary, it can be concluded that the 5. EN ISO 16150:2012 Water quality. environmental situation after criteria of Regulation Guidance on pro-rata Multi-Habitat sampling of N4 in the upper reaches of the River Iskar is benthic macro-invertebrates from wadeable rivers. acceptable. Despite this, deterioration of ecological 6. Gardanov T., K. Ivanov, B. Russev, D. quality in lower part of catchment was reported. This Pechinov, P. Petrov, V. Semov, E. Kabakchieva, is probably due to settlements and agriculture Study on self-purification processes in Iskar and activities and river fragmentation (for fishes). The river valleys, Trudove NIVKST, Vol. VII (2), growing tourist activity in the region may increase 1971, 17-37. (In Bulgarian). the anthropogenic pressure on these vulnerable 7. Kanev E., M. Mitankina, K. Ignatov, E. mountain ecosystems, not only by organically Uzunova, Longitudinal connectivity for fish loaded water inflow but also by fishing activities. migration in Upper Reaches of the Iskar River and The latest may have great impact in small mountain prioritisation of the restoration measures, Proc. of rivers. A more precise analysis of the dynamic the 41st International Association for Danube changes in the ecological quality according to the Research (IAD) Conference - 60 years of IAD criteria laid down in Regulation N4 is not possible. existence, Tributaries as key elements in sustainable This is due to the limitations in the accepted management of the Danube River Basin,Sofia, methodologies used in characterization of surface , 2016, 13-16. waters. For instance, BI used scores (1 to 5) not 8. Kenderov L., I. Yaneva, Ecological numbers or percentages (EQR), and TTN is characteristics of the Iskar River catchment, Biotechnol. dependent on the natural dynamics of communities. & Biotechnol. Eq, 23, 2009, 276 – 280. In order to adequately assess of the ecological 9. Kenderov L., I. Yaneva, M. Pavlova, situation on the basis of the fish fauna, it is Ecological Assessment of the Upper Stretch of the necessary to increase the number of investigated Iskar River Based on Selected Biological Parameters sites and the frequency of the monitoring events. in Conformity with the Wаter Frame Directive Acknowledgements: This study is supported by 2000/60/EU, Acta zoologica bulgarica, Suppl. 2, Sofiyska Voda AD, a partner in the project “Live 2008, 255-268. water”. 10. Kottelat M., J. Freyhof, Handbook of European Freshwater Fishes, Edit. Publications REFERENCES Kottelat, Cornol, Switzerland, 2007, 646 p. 1. CEN document, Water quality – Sampling of 11. Mihov S., Development of fish based index fish with electricity, CEN/TC 230, Ref. No. EN 14011: for assessing ecological status of Bulgarian rivers 2003, E. 16 p. (BRI), Biotechnol. & Biotechnol. Equipment, 24 2. Cheshmedjiev S., P. Ivanov, R. Mladenov, G. (online edition), 2010, 248-256. Gecheva, S. Mihov, D. Todorov, Development of a 12. Parvanov D., Y. Topalova, I. Janeva, classification system for assessment the ecological status Bacterial and Macroinvertebrate Community and ecological potential of the identified types of surface Structure Analyses in Relation to Chemical waters (rivers and lakes) on the territory of the Republic Variables of Water and Sediments in the Central of Bulgaria (according the typology “B”), Final Report Part of Iskar River Basin, Ecological Engineering of the project No. 6/27.03.2009b with Basin and Environmental Protection, 6 (1), 2007, 21-30. Directorates: West Aegean Sea Region, Danube Region, 13. Paspalev G., Ts. Peshev, A contribution to Region and East Aegean Sea Region. the knowledge of the ichthyofauna of Iskar River, Consortium Biomonitoring, Sofia, 2010, 120 p. (In Annulaire de l’Universite de Sofia, Faculte de Bulgarian). Biologie, Geologie et Geographie, 48 (1), 1955, 1 – 3. Cowx I., J. Harvey, R. Noble, D. Nunn, 39. (In Bulgarian). Establishing survey and monitoring protocols for the 14. Pavlova M., L. Pehlivanov, Еcological assessment of conservation status of fish populations in assessment of Palakariya River (South-West river Special Areas of Conservation in the UK, Aquatic Bulgaria, Danube River basin) based on fish and Conserv. Mar. Freshw. Ecosyst. 19, 2009, 96-103. macrozoobenthic communities. – In: Proceedings of 4. EN ISO 10870:2012 Water quality - Workshop on Ecology – 2009 (Chankova S., S. Guidelines for the selection of sampling methods Gateva, G. Yovchev, N. Chipev, Eds.), 2009, 74-81.

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15. Pehlivanov, L., M. Pavlova, M. Vassilev, A. 18. Uzunova E., E. Kanev, T. Stefanov, Spatial Apostolou, B. Velkov, Ecological quality Variation in the Abundance and Population assessment of salmonid rivers in Bulgaria using Structure of Bullhead Cottus gobio (Pisces: ichthyologic parameters, Acta zoologica bulgarica, Cottidae) from the Iskar River Basin (Danube River Suppl. 4, 2012, 97-203. Drainage, Bulgaria): Implications for Monitoring 16. Russev B., Influence of industrial and Conservation, Acta Zool. Bulg, 2017, (in press). wastewaters on life in Arda River and Iskar River, 19. Yaneva I., B. Rusev, Saprobiological state Priroda, Vol.4, 1959, 51-55. (In Bulgarian). of Iskar River after beginning of exploitation of 17. Russev B., Saprobiological assessment of some Sofia wastewater treatment plant, Hydrobiology- Bulgarian rivers, Bulletin of Zoological Institute with BAS, Vol.34, 1989, 3-17. Museum, BAS, 27, 1968, 141-156. (In Bulgarian).

ЕКОЛОГИЧНО СЪСТОЯНИЕ НА ГОРНОТО ТЕЧЕНИЕ НА РЕКА ИСКЪР И НЯКОИ ПРИТОЦИ СЪГЛАСНО НАРЕДБА Н4, СПОРЕД МАКРОЗООБЕНТОСА И РИБНАТА ФАУНА

Любомир Кендеров, Димитрий Дашинов, Емил Кънев, Любомира Любомирова, Елиза Узунова

Резюме: Представена е актуална информация за екологичното състояние на горното поречие на р. Искър, включително някои нейни притоци. Извършена е екологична оценка съгласно РДВ и националното законодателство (Наредба Н4). За изчисляване на базираните на риби индекси (BRI и TRI) са използвани следните ихтиологични параметри: обща численост и биомаса, численост и биомаса на Salmo trutta, срещаемост на интродуцирани видове, чувствителни и мигранти. За определяне на индексите BI и TTN е използван таксономичния състав на макрозообентоса. Екологичната ситуация на река Искър и притоците е определена като „отлична“ според индексите на макрозообентоса и „умерена“ до „лоша“ според базираните на риби индекси. Наблюдава се известно влошаване в най-долните речни участъци. Ключови думи: екологично състояние, ихтиофауна, макрозообентос, река Искър

Assistant Professor Lyubomir Kenderov, PhD гл. ас. д-р Любомир Кендеров, Sofia University „St. Kl. Ohridski”, Faculty of Софийски университет „Св. Климент Охридски”, Biology, Department of General and Applied Биологически факултет, Катедра „Обща и Hydrobiology приложна хидробиология” Address: 1164 Sofia, 8, Dragan Tzankov Blvd., room адрес: гр. София (п. код 1164), бул. „Д. Цанков” 8, 168 каб. 168 tel.: 02/8167343 тел.: 02/8167343 e-mail: [email protected] ел. поща: [email protected]

Dimitriy Dashinov, PhD student докторант Димитрий Дашинов, Sofia University „St. Kl. Ohridski”, Faculty of Софийски университет „Св. Климент Охридски”, Biology, Department of General and Applied Биологически факултет, Катедра „Обща и Hydrobiology приложна хидробиология” address: 1164 Sofia, 8, Dragan Tzankov Blvd., room адрес: гр. София (п. код 1164), бул. „Д. Цанков” 8, 168 каб. 168 tel.: 0879206804 тел.: 0879206804 e-mail: [email protected] ел. поща: [email protected]

Emil Kanev, PhD student докторант Емил Кънев, Sofia University „St. Kl. Ohridski”, Faculty of Софийски университет „Св. Климент Охридски”, Biology, Department of General and Applied Биологически факултет, Катедра „Обща и Hydrobiology приложна хидробиология” Address: 1164 Sofia, 8, Dragan Tzankov Blvd. адрес: гр. София (п. код 1164), бул. „Д. Цанков” 8 e-mail: [email protected] ел. поща: [email protected]

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Lyubomira Lyubomirova, PhD student докторант Любомира Любомирова, Sofia University „St. Kl. Ohridski”, Faculty of Biology Софийски университет „Св. Климент Охридски”, Department of Zoology and Anthropology Биологически факултет, Катедра „Зоология и Address: 1164 Sofia, 8, Dragan Tzankov Blvd., антропология” e-mail: [email protected] адрес: гр. София (п. код 1164), бул. „Д. Цанков” 8 ел. поща: [email protected]

Associated Professor Eliza Uzunova, PhD доц. д-р Елиза Узунова, Sofia University „St. Kl. Ohridski”, Faculty of Софийски университет „Св. Климент Охридски”, Biology, Department of General and Applied Биологически факултет, Катедра „Обща и Hydrobiology приложна хидробиология” Address: 1164 Sofia, 8, Dragan Tzankov Blvd., room адрес: гр. София (п. код 1164), бул. „Д. Цанков” 8, 162B каб. 162Б Tel.: 02/8167315 тел.: 02/8167315 e-mail: [email protected] ел. поща: [email protected]

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