Microbiological water quality of the River Danube (km 2581 - km 15): Longitudinal variation of pollution as determined by standard parameters

Gerhard G. Kavka1, Georg D. Kasimir2, and Andreas H. Farnleitner3

Keywords: Microbiological water quality, Large Rivers, faecal pollution

Introduction The examination of microbiological river water quality according to technical standards is obligatory for use-related aspects such as for drinking water production, irrigation or recreation. Microbiological data for total coliforms, faecal coliforms (representing Escherichia coli as predominant species), intestinal enterococci (faecal streptococci) and heterotrophic plate count (colony count) were collected during the Joint Danube Survey (JDS) 2001, organised by the International Commission for the Protection of the Danube River (ICPDR), along the longitudinal stretch of the River Danube from the upper section (km 2581) to the Black Sea and in major tributaries. Data were used for the assessment of microbial pollution along the course of this large river and to establish a microbiological water quality map of the investigated river basin. In addition observed microbiological data were compared with data from the Danube survey 1988, organised by IAD. Microbial pollution due to anthropogenic impacts, especially from large urban settlements, were analysed within defined sections of the River Danube and furthermore compared to biological and chemical data. Indicator bacteria and analysis methods Faecal indicator bacteria like total coliforms, faecal coliforms (thermotolerant coliforms), E. coli and intestinal enterococci (faecal streptococci) are excreted by humans and warmblooded animals, pass sewage treatment plants to a great amount and survive for a certain time in the aquatic environment (Kavka & Poetsch 2002). E. coli and faecal coliforms are the best indicators for the assessment of recent faecal pollution, mainly caused by raw and treated sewage and diffuse impacts e. g. from farm land and pasture. E. coli and faecal coliforms indicate also the potential presence of pathogenic bacteria, viruses and parasites (Kavka & Poetsch 2002). Detailed knowledge of faecal pollution in aquatic environments is crucial for watershed management activities in order to maintain safe waters for recreational and economic purposes (Farnleitner et al. 2001). The concentrations of heterotrophic bacteria (heterotrophic plate count 22C, colony count 22C) correspond commonly with contamination by organic matter (Kohl 1975). Analysis of variations of indicator bacteria in the longitudinal stretch of the Danube River and major tributaries were carried out by uniform standard methods in a laboratory on-board to receive comparable results according to ISO 9308-1 1990, EN ISO 7899-2 2000, EN ISO 6222 1999, DEV K5 1971, DIN 38411-5 1983 (for more details see Kavka & Poetsch 2002).

Federal Ministry of Agriculture, Forestry, Environment and Water Management, Federal Agency for Water Management, A-3252 Petzenkirchen, Austria
2 3

HYDRA-Institute sterreich, Himmelstrasse 42, A-1190 Vienna

Institute of Chemical Engineering, Department for Applied Biochemistry and Gene Technology, University of Technology, Getreidemarkt 166-9, A-1060 Vienna

415

Variations of microbiological parameters along the longitudinal course of the Danube River and in major tributaries The Joint Danube Survey (JDS) 2001 included 98 sampling sites from Germany (Neu-Ulm, stream-km 2581) to the Black Sea (Sulina arm, stream-km 15). The variation of the microbiological results of all sampling sites along the flow stretch of the River Danube is presented exemplarily by the parameter faecal coliforms (Fig. 1). Tributaries are symbolised as small bars. The target value for the concentrations is marked on the chart.
[CFU/ 100ml] 100 000 000 10 000 000 1 000 000 100 000 10 000 1 000 100 10 1 2600 2400 2200 2000 1800 1600 1400 1200 1000 800 [Danube km]

Faecal Coliforms
Rackeve-S.-Arm Russenski Lom Moson-Arm Arges

Velika Morava

600

400

200

Siret Prut 0

Fig. 1: Variation of faecal coliforms along the longitudinal course of the Danube River; small bars = tributaries; dotted line = target value; CFU=colony forming units Total coliforms varied from 60 to 75 000, faecal coliforms from 20 to 41 000, intestinal enterococci from 5 to 2 200 CFU/ 100ml. The target values amount to 10 000/100ml (= imperative value acc. to EU-Bathing water quality directive 76/160 EEC), 1000/100ml and 400/100ml (= target values acc. to the EU-Bathing water quality directive 06/7/EEC) respectively (see table 2). In about 42% of all Danube sampling sites including arms and in 53% of the tributaries no compliance with the requirements concerning faecal indicator bacteria were found. In the Danube arms Moson Danube arm, Rackeve-Soroksar Danube arm and the tributaries Ipoly, Russenski Lom, Arges, Siret and Prut" a great lot of faecal indicator bacteria were detected. Heterotrophic plate counts varied from 240 to 54 000 per 1ml in the Danube River. The target value is 10 000 colonies per 1ml water in consideration to Kohl (1975). In about 13% of sampling sites of the Danube River more colonies than acceptable were isolated. In the tributaries Drava, Russenski Lom and Arges the highest colony counts (maximum 1 400 000 /ml) occurred. 42% of the tributaries did not comply with the requirements. The concentration ranges of indicator bacteria analysed during JDS 2001 and IAD Survey 1988, arranged by the International Association for Danube Research, are presented in table 1. Both investigations were carried out along the Danube River by uniform methods. JDS took place in August/September 2001, the IAD survey in March 1988. The ranges of results of the two surveys are comparable, although the date sets has been recovered from different seasons.

416

Table 1: JDS 2001 and IAD-Survey 1988; concentration ranges of bacteriological parameters in the longitudinal course of the Danube River; JDS = Joint Danube Survey ICPDR; IAD-Survey = Danube survey of the International Association for Danube Research Danube Survey No of measured sites Total coliforms Faecal coliforms Intestinal enterococci Plate count 22C 100ml 100ml 100ml 1ml
2 2

JDS 2001 79

IAD-Survey 1988 25

Concentration ranges 4 2 4 6.0x10 7.5x10 * 4.0x10 5.3x10 ** 2.0x10 4.1x10 * 5.0 - 2.2x103* 2.4x10 5.4x10 *
2 4 4

3.0x10 2.3x10 ** 1.0x10 - 2.7x10 *** 1.1x10 1.2x10 **

3 5 1 3

*Kavka & Poetsch (2002); **Albinger (1988); ***Trzilova & Miklosovicova (1988) Microbiological Water Quality Map Microbiological water quality of River Danube and its tributaries has been assessed by the concentrations of standard microbiological parameters, classified by 5 quality classes (table 2) and represented in a microbiological map (fig. 2). Table 2: Class limit values for microbial pollutions of rivers assessed by bacteriological standard parameters according to Kohl (1975, modified), Kavka & Poetsch (2002, modified) and in consideration of EU-Bathing water quality directive 2006/7/EEC (target values for E. coli and enterococci concerning a good microbiological bathing water quality = target values in this table) and directive 76/160 EEC (imperative value for total coliforms concerning bathing water quality = target value in this table); TV=Target Value; FS= faecal streptococci a) faecal pollution, b) pollution by easily degradable organic matter a) Classification of faecal pollution Parameter Escherichia coli (Faecal Coliforms*) Intestinal Enterococci (FS) Total Coliforms Faecal pollution in 100ml water in 100ml water in 100ml water Class I little < 100 < 40 < 500 II (TV) moderately > 100 - 1 000 > 40 - 400 > 500 -10 000 III critical > 1 000 - 10 000 > 400 - 4 000 > 10 000 - 100 000 IV strongly > 10 000 - 100 000 > 4 000 - 40 000 > 100 000 - 1000 000 V excessively > 100 000 > 40 000 >1000 000

* Faecal coliforms are mainly represented by Escherichia coli as predominant species

417

b) Classification of organic pollution Parameter Heterotrophic Plate Count 22C Organic pollution in 1 ml water CLASS I little < 500 II moderately > 500 - 10 000 III critical > 10 000 - 100 000 IV strongly > 100 000 -750 000 V excessively > 750 000

Figure 2 presents a map of faecal pollution of the Danube River at 98 measured sites during JDS 2001. 5 Classes of faecal pollution were assessed by the concentrations of faecal coliforms and additionally by the concentrations of intestinal enterococci (faecal streptococci) (acc. to table 2). Because one sample per site was taken, direct assessment of the bathing water quality at the sampling locations cannot be given. The EU- bathing water quality directive requires replications and their respective percentiles. The microbiological samples were taken from the middle of the River Danube. Additional investigations from both river banks would allow a better detecting of the impact of tributaries, raw sewage or waste water treatment plants. In the German and Austrian section of the Danube River (km 2581 1880) faecal indicator bacteria indicated a little to moderate faecal contamination and organic pollution in the Danube and the tributary Inn (classes I and II) (figure 2). Only at one station in Hainburg, Danube km 1881, concentrations of faecal indicator bacteria were relative high (critical faecal pollution, class III). Influences from local contamination by the settlement Hainburg, from the tributary Schwechat River, receiving treated waste water, and probably from the sewage treatment plant of Vienna, are supposed to be responsible. The tributaries Schwechat and Morava River were critical polluted by indicator bacteria. Results were corresponding with data received during the Danube survey From River Rhine to the Hungarian stretch of River Danube (1998). Sampling sites along the Danube course and in side-arms in Slovakia, Hungary, Croatia, Serbia, and Romania upstream Iron Gate I (stream km 1880 956) were moderately to strongly faecal polluted (classes II, III, IV) (figure 2). Plate counts corresponded with moderate to critical organic pollutions (classes II, III). An influence of the city Bratislava seemed probably. The impact of Budapest was significant from the bacteriological point of view. A marked increase of bacterial pollution was observed downstream from Budapest. The City Belgrade probably caused a marked increase of pollution indicated by standard bacteriological parameters. An influence of the Morava River seemed probably. The tributary Vah contained bacterial pollution indicators only in small amounts. In the tributary Hron the bacteria were enumerated in higher amounts corresponding with a critical pollution (class III). The Ipoly River was classified as critical organic (class III) and strongly faecal polluted (class IV). The influence of the bacterial contaminated river Drava seemed remarkable (class III). Downstream Novi Sad critical bacterial concentrations occurred (class III). In the rivers Sio and Tisa a good bacteriological water quality was observed during investigation time. The tributary Sava was moderately contaminated by indicator bacteria (class II). The River Velika Morava contained increased concentrations of faecal coliforms. The Danube reach in Romania from stream-km 956 (downstream Iron Gate I) 537 was the section with the best bacteriological water quality during the sampling period. Plate counts

418

JDS 22 JDS 21 Sap Outlet

svnyrr 1812 1812

UA

JDS 02 JDS 12
Klosterneuburg 1942 Bratislava 1869

JDS 04 JDS 23
Medvedov 1806 Moson arm end 1794

JDS 17

D
JDS 16 JDS 14
Wildungsmauer 1895 tr. Morava 1880

Kelheim 2412

Kachlet 2233

JDS 01 JDS 08 JDS 10

Morava

JDS 06

Neu Ulm 2581

Altm

hl

Jochenstein 2200

SK
1708

Le c h

Isa r

JDS 03

Asten 2120

Ybbs 2061

JDS 29 Hron JDS 30 JDS 26 tr. 1716 tr. Ipoly

JDS 251766

tr. Vah

Geisling 2358

Inn

Bratislava Vienna JDS 11

Vah
Szob 1707 1692

Komarno 1768

JDS 32 us. Szetendre Island JDS 31

MD
Kishinev

s Ille

JDS 33

Ris

JDS 05
Enns
Greifenstein 1950

Munich
JDS 09 JDS 13
Raba

tr. Inn 2221 Wallsee 2095 tr. Schwechat 1913

JDS 07 JDS 27 JDS 28

Szony 1761 Esztergom 1719

Szetendre arm start JDS 34 1692 us. Budapest 1659

Aschach 2165

Budapest
JDS 36
Soroksar arm start 1642

A
us. Morava 1881 Gabcikovo res. entr. 1856 Dunafldvar 1560

JDS 15 JDS 20
Gabcikovo res. 2 1846

JDS 37
ds.Budapest 1632

JDS 35
Szetendre arm end 1658

JDS 18 JDS 39 JDS 41

Paks 1533 Soroksar arm end 1586

JDS 40

JDS 38

JDS 94
tr. Prut 135

JDS 96 JDS 95
t Pru

JDS 19
Gabcikovo res. 1 1852 tr. Sio 1497

H
JDS 43 JDS 64 JDS 63 JDS 52
ds. Novi Sad 1252 Starapalanka - Ram 1077 Banatska Palanka / Bazias 1071 Baja 1481

Tass 1586

t Sire

JDS 42 JDS 44
Hercegszanto 1434

JDS 93

Reni 130 tr. Siret 154

Ismail

Chilia arm (Vilkova) 56

BLACK SEA
JDS 97 JDS 92 JDS 82
ds. Jantra 532 Braila 167 Sulina arm 12

JDS 45 JDS 65
Irongate reservoir (Golubac / Koronin) 1040

JDS 46
us. Drava 1384

RO

Zagreb

Batina 1429

HR
JDS 47
Tisa

Dra va

JDS 48
ds. Drava 1367 tr. Tisa 1215

JDS 54 JDS 66 JDS 55

Irongate reservoir 954 ds. Tisa 1202

JDS 98 JDS 89 JDS 80

Chiciu / Silistra a 375 mit Bucurest Ialo

tr. Drava 1379

Tamis

JDS 91 JDS 75

St. Gheorghe arm 64 Giurgeni 235

JDS 49
Dalj 1355

Danube
JDS 53 JDS 50
Sa va

JDS 90
ds. Iskar 630 Svishtov 550

JDS 57

Grocka 1132

Morava

ra

Isk ar

Vit

Jant

Lo m

419
Belgrade
JDS 56
tr. Sava 1170 us. Tisa 1216

JDS 67
Vrbica / Simijan 924

0 JDS 62 JDS 59 JDS 61 JDS 69

tr. Timok 845 tr. Morava 1103 1097 Ilok / Backa Palanka JDS 51 1300 us. Novi Sad 1262

50

100

150

250

250 km

JDS 87 JDS 73 JDS 77

us. Iskar 640 tr. Olt 605 tr. Arges 432

Arge s

us. Cernavoda 295

JDS 68
us. Timok 849
Jiu

JDS 88
ds. Arges

JDS 86 429
Olt

BIH
JDS 60
us. Morava 1107

us. Pancevo 1161

us. Arges Ru 434 s. Lo m

am Os ta os Og

JDS 85 JDS 70
Novo Selo 834

t Sk

JDS sampling point Tributary

JDS 58

Sarajevo
1151

JDS 74
tr. Iskar 637

JDS 83 JDS 79
Nikopol 579 us. Ruse 500

ds. Ruse 488

JDS 84

Tr. = tributary

FRY

JDS 71
Calafat 795

JDS 72
ds. Kozloduy 685 us. Olt 606

JDS 76 ds. Olt

JDS 78
602

JDS 81
tr. Jantra 537

tr. Russenski Lom 498

us. = upstream

ds. = downstream

Sofia

BG

JDS 01 Neu Ulm 2581

JDS point number Location Danube km

available within

Figure 2: Microbiological map of faecal pollution; Classes of faecal pollution assessed by the concentrations of faecal coliforms and additionally concentrations of intestinal enterococci (faecal streptococci); classification system see table 2; blue coloured symbols: class 1 - little faecal polluted; green: class II- moderately; yellow: class III-critical; orange: class IV-strongly; red: class V-excessively faecal polluted; Base map acc. to Vogel & Pall (2002).

and faecal bacteria indicated little to moderate pollution by organic matter and faeces respectively (classes I and II). The tributaries Timok, Jantra and Olt were low contaminated. Sampling sites at the River Danube in Romania/ Bulgaria and in the Danube Delta (km 537 15) were classified as moderate to strongly faecal (classes III IV) and moderate to critical organic polluted (classes II, III). Russenski Lom and River Arges were the most contaminated tributaries, which has been investigated during JDS and classified as excessively faecal and organic polluted (class V). The rivers Siret (class III) and Prut (class IV) were significant pollution factors for the River Danube. Data of selected microbiological parameters (heterotrophic plate count, total and faecal coliforms, enterococci), physico-chemical parameters (dissolved oxygen, organic nitrogen, ammoniacal- and nitrate-nitrogen, ortho-phosphate-phosphor) and biological parameters (Saprobic index - Macrozoobenthos) were comparatively classified along the Danube River. All data were made kindly available by partners of JDS 2001 project. In more than 15% of all sampling locations (river sites, arms, tributaries) microbiological parameters alone indicated remarkable anthropogenic impacts characterised by high bacterial concentrations with non compliance to the target values. This fact supports and vindicates the application of microbiological parameters in monitoring programmes. It is well-known, that - although biological and chemical water quality is acceptable, bacteriological parameter can be detected in critical concentrations (Baumann & Popp 1991). Summary Bacteria are ideal sensors for the indication of microbial pollution of surface water bodies because of their fast response to changing environmental conditions. Faecal coliforms E. coli and intestinal enterococci (faecal streptococci) are good indicators for the assessment of faecal pollution mainly caused by raw and treated sewage and diffuse impacts from farmland and pastures, also indicating the potential presence of pathogenic bacteria, viruses and parasites. The concentrations of heterotrophic bacteria (plate count) correlate commonly to organic pollution. For monitoring the quality of river water intended for the abstraction of drinking water, irrigation and bathing, the examination of these microbiological standard parameters is made obligatory by legislation. Since Danube countries normally use different methods of microbiological analysis, JDS offered a unique opportunity to obtain comparable results from Neu Ulm (Germany) to the Black Sea. In JDS 2001 standard microbiological parameters were analysed on board by using uniform methodology for all sampling sites. The evaluation of microbiological results located the highest pollution in the tributaries (Russenski Lom, Arges, Siret and Prut in particular) and in the side arms (Moson arm, RackeveSoroksar arm). Lower bacterial pollution could be observed in the Upper Danube reach as well as in and downstream of the Iron Gate reservoir. Higher levels of faecal pollution were found in the middle part of the Danube, particularly downstream of major cities (Budapest, Beograd) as far as 1.100 river km and again in the Lower Danube from stream-km 500 to the Danube Delta. The main sources affecting the microbiological water quality were raw sewage, discharges from wastewater treatment plants, impaired tributaries and impact by diffuse sources. The fact that in a remarkable number of sampling sites microbiological parameters alone indicated anthropogenic impacts supports and vindicates the application of microbiological parameters in monitoring programmes. Knowledge on microbial pollution in lotic aquatic environments appears essential for decision makers in order to take appropriate measures which result in acceptable river water quality and compliance with national and international quality standards and directives.

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