Submitted:
03 September 2025
Posted:
08 September 2025
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Abstract
Keywords:
- Ceratophyllum demersum and Lemna minor exhibit a strong capacity for phytoextraction of heavy metals and boron.
- Eichhornia crassipes and Pistia stratiotes facilitate the accelerated biodegradation of petroleum hydrocarbons and demonstrate high efficiency in sorbing surfactants.
- Typha latifolia and Phragmites australis contribute to denitrification, stabilization of bottom sediments, and metal precipitation.
Introduction
Historical Sources of Pollution in the Ilek River Basin
Rationale for the Use of Macrophytes and Bioindication in the Ilek River
Pollution Challenges of the Ilek River and the Relevance of Macrophyte Selection
Mechanisms of Action and Rationale for Macrophyte Selection
Environmental Incidents of 2025 and Associated Bio-Risks
Transboundary Context, Sources, and Hydrochemical Profile
State of Research and Methodological Gaps
Justification for the Selection of Bioindicators and Nature-Based Measures
International Context and Regulatory Frameworks
Scientific Rationale for the Selected Approach
Need for an Integrated Assessment
- Bioindication — calculation of saprobity indices and assessment of macrophyte community status, accounting for their capacities for bioaccumulation and biodegradation;
- Chemical analysis — determination of key contaminants (Cr, B, Pb, Fe, Mn, petroleum hydrocarbons, surfactants) and routine metrics (BOD₅, COD);
- Spatial analysis — identification of priority zones for remediation and the deployment of nature-based solutions (constructed wetlands, buffer strips, phytoremediation).
Research Objectives
- Conduct field surveys to quantify physicochemical and biological parameters of the Ilek River, with emphasis on Cr, Pb, Fe, Mn, B, petroleum hydrocarbons, surfactants, COD, and BOD₅.
- Characterize the species composition and spatial distribution of macrophyte communities within the channel and riparian zones, and compute the saprobity index (S) for indicative diagnosis of pollution.
- Based on the literature, evaluate the effectiveness of key macrophyte taxa (Ceratophyllum demersum, Lemna minor, Eichhornia crassipes, Pistia stratiotes, Typha latifolia, Phragmites australis) in removing priority contaminants.
- Compare, using the obtained data, the performance of individual species and their mixtures in terms of removal rates and overall pollutant reduction.
- Provide practice-oriented recommendations for integrating phytoremediation technologies into monitoring programs and into the management of anthropogenic pressures on the Ilek River.
2. Materials and Methods
2.1. Study Area
- Channel width: 20–70 m (average ~35 m)
- Depth: 0.8–3.5 m
- Flow velocity: 0.2–0.6 m/s
- Biotope types: main channel sections with hard and silty substrates, backwaters, shallow zones with macrophyte vegetation, and low-flow areas
2.2. Field Survey Design

- Reconnaissance survey with recording of biotope types and physico-morphological features.
- Photographic documentation of riparian vegetation and visible signs of pollution.
- Water sampling at sites with the highest saprobity index (S) values or pronounced visual indicators of contamination.
- Laboratory analysis of hydrochemical parameters.
- Calculation of the macrophyte-based saprobity index (S) and preparation of a spatial distribution map.
- Comparative analysis of bioindication results and chemical monitoring data.
2.3. Hydrochemical Parameters of Monitoring
| Parameter | Analytical Method | Units | Standard/Regulatory Document |
| BOD₅ | Titrimetric, manometric | mg O₂/L | GOST 31859–2012 |
| COD | Dichromate method | mg O₂/L | GOST 31861–2012 |
| Oil products | IR spectrophotometry | mg/L | GOST 31957–2012 |
| Cr, Pb, Fe, Mn, B | Atomic absorption spectrometry (AAS) | mg/L |
2.4. Methodology for Calculating the Saprobity Index (S)
- si — the saprobic index value of species i (based on macrophyte saprobity reference manuals);
- hi — the relative coverage of species i at the sampling site (%);
2.5. Water Sampling at the Most Contaminated Sites
- Biochemical Oxygen Demand (BOD₅): Water samples were collected in 250 mL glass bottles with ground-glass stoppers. The analysis was conducted according to RD 52.24.420-2005, by measuring the difference in dissolved oxygen concentrations before and after 5-day incubation at 20 °C in the dark.
- Chemical Oxygen Demand (COD): Samples were collected in glass bottles, and COD was determined photometrically using the dichromate method in accordance with PND F 14.1:2:4.153-99.
- Petroleum Hydrocarbons: Samples were collected in glass bottles pre-rinsed with hexane. The analysis was carried out using infrared (IR) spectrophotometry according to PND F 14.1:2:4.128-97.
- Surfactants (Synthetic Detergents): Samples were collected in clean glass containers. The determination was performed by the photometric method based on the formation of a complex with methylene blue, in accordance with RD 52.24.417-2005.
2.6. Laboratory Analysis of Heavy Metals
2.7. Experimental Workflow

2.8. Statistical Processing
3. Results and Discussion
| Photo No. | Dominant macrophytes (Latin) | Possible pollution indicator | Likely pollution source |
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Phragmites australis | Dense reed stands, eutrophication | Point: untreated sewage; Diffuse: stormwater runoff |
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Phragmites australis | Littoral expansion, nutrient accumulation | Point: storm drains; Diffuse: lawn runoff |
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Phragmites australis | High biomass, nutrient saturation | Point: sewage; Diffuse: fertilizers |
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Phragmites australis | Early-stage littoral growth, clear water | Point: drainage; Diffuse: erosion |
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Phragmites australis, Salix spp. | Mixed riparian vegetation, good oxygen regime | Point: domestic wastewater; Diffuse: low flow |
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Phragmites australis, Carex spp. | Restricted flow, organic matter accumulation | Point: storm drains, litter; Diffuse: nutrient deposition |
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Phragmites australis | High biomass, eutrophication | Point: discharges; Diffuse: fertilizers |
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Phragmites australis | Domestic pollution | Point: storm drains; Diffuse: lawns |
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Phragmites australis, Salix spp. | Good aeration, potential accumulation | Point: domestic; Diffuse: erosion |
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Phragmites australis | Eutrophication, weak flow | Point: discharges; Diffuse: agricultural runoff |
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Phragmites australis | Organic matter accumulation | Point: litter; Diffuse: fertilizers |
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Phragmites australis | Nutrient accumulation, high saprobity | Point: drainage; Diffuse: agricultural runoff |
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Phragmites australis under Salix spp. canopy | Anaerobic processes in sediments | Point: groundwater seepage; Diffuse: erosion |
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Phragmites australis, Salix spp. | Early-stage eutrophication | Point: sewage; Diffuse: stormwater runoff |
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Phragmites australis | Transition to algal bloom | Point: sewage; Diffuse: croplands |
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Phragmites australis, Salix spp. | Moderate saprobity | Point: drainage; Diffuse: erosion |
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Phragmites australis, Salix spp. | Buffer filtration with excess nutrients | Point: discharges; Diffuse: fertilizers |
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Phragmites australis | Eutrophication indicator | Point: storm drains; Diffuse: precipitation |
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Phragmites australis | High macrophyte productivity | Point: discharges; Diffuse: fertilizer runoff |
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Phragmites australis, Salix spp. | Filtration and sedimentation zone | Point: domestic wastewater; Diffuse: croplands |
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Phragmites australis with understory | Stable littoral zone | Point: drainage; Diffuse: precipitation |
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Phragmites australis | Overenrichment, algal bloom onset | Point: storm drains, sewage; Diffuse: fertilizer runoff, slope erosion |
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— | Organic pollutants, surfactants, oil products | Point: faulty wells; Diffuse: agricultural seepage |
- Minimum S values (2.4–2.6) were recorded at sites dominated by Phragmites australis with the presence of shrubs. These values correspond to the β-mesosaprobic zone, indicating moderately polluted but still relatively clean waters.
- Elevated S values (S ≥ 3.0) were observed in areas with visible signs of eutrophication, such as the presence of foam and litter. These values correspond to the α-mesosaprobic zone, reflecting significant organic pollution.
- Maximum S values (up to 3.2) were typical of sites near urban effluents and technical wells, indicating strong anthropogenic impact.
Bioindication as an Integrative Approach
Managing Anthropogenic Pressure on the Ilek River
- The overall pollution pattern: how water quality changes from the headwaters to the mouth.
- Local pollution sources: specific sites with maximum contamination.
- Effectiveness of mitigation measures: in the long term, regular monitoring of the profile will allow an assessment of how the river’s condition responds to conservation activities.

Analysis of the Saprobity Profile of the Ilek River
Relationship Between Macrophytes and Hydrology
The Need for Additional Indicators in Polluted River Sections
Rationale for Additional Indicators
Case Study: Localized Pollution Hotspots
Integrated Approach to Assessment
Analytical Summary
Integrated Interpretation
- Sites with S ≥ 3.2 are critical and require urgent intervention.
- The saprobity index shows a strong correlation with BOD₅ and petroleum hydrocarbons (r > 0.85, p < 0.05, n = 7).
- The Pantle–Buck method, adapted for macrophytes, is justified as a reliable tool for the early detection of pollution hotspots and for prioritizing sites for monitoring and remediation.

-
Positive influence of BOD₅ and petroleum products (positive coefficients for BOD₅ and petroleum hydrocarbons) confirms that:
- ○
- the organic load (BOD₅) is directly reflected in the saprobity index;
- ○
- petroleum hydrocarbons intensify eutrophication and biological stress, leading to an increase in S values.
- Hexavalent chromium (Cr) has the largest coefficient (15.536), which highlights its critical role as a toxicant, especially in combination with other pollutants. It affects biota both directly and indirectly, by accelerating the degradation of macrophyte communities and altering saprobic structure.
-
Negative influence of boron and COD (negative coefficients) may be associated with their relative biological inertness or compensatory effects:
- ○
- COD also accounts for hardly oxidizable substances that are not always bioavailable;
- ○
- boron, at low concentrations, may play a role as a micronutrient, while its excess does not necessarily lead to an immediate increase in saprobity.
Ecological and Methodological Implications
- more accurate and ecologically meaningful diagnostics of water body status;
- predictive capacity to assess pollution levels based on readily available chemical-analytical data;
- scientifically grounded recommendations for monitoring systems and environmental management decisions.
- the index increases with higher BOD₅ and petroleum hydrocarbons, which is logical since they directly reflect organic pollution;
- the index decreases with higher chromium and boron, likely due to their toxic effects that suppress biotic development, thereby limiting eutrophication and instead reducing biodiversity.
Practical Application


- in areas under anthropogenic pressure from agricultural and industrial zones,
- where point and diffuse discharges are observed,
- and where constant laboratory monitoring is not feasible—calculation of S as an integral indicator is sufficient.
| Macrophyte | Share (%) | Justification |
|---|---|---|
| Eichhornia crassipes | 25 | High efficiency in removing oil products and Cr(VI); forms stable microbial communities active in hydrocarbon biodegradation. Dominant under conditions of organic and heavy metal pollution. |
| Pistia stratiotes | 15 | Supports Eichhornia, enhances oil sorption, effective in stagnant zones, improves phytobarrier structure. |
| Ceratophyllum demersum | 20 | Removes BOD and boron, stabilizes bottom zones, participates in nitrification, effective under high organic load. |
| Lemna minor | 10 | Fast-growing primary filter, effective in removing surfactants, boron compounds, and BOD; forms the first protective surface layer. |
| Typha latifolia | 20 | Coastal filter, stabilizes flow, promotes Cr precipitation and COD removal; creates reducing conditions in the rhizosphere. |
| Phragmites australis | 10 | Slow but resilient biofilter; completes purification from Cr and phenols, highly compatible with Typha. |

Conclusions
Key Findings of the Research
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| Saprobity Zone | S Range | Ecological Characterization |
|---|---|---|
| Oligosaprobic | 1.0–2.0 | Clean water |
| β-Mesosaprobic | 2.0–2.5 | Moderate pollution |
| α-Mesosaprobic | 2.5–3.0 | Elevated organic load |
| Polysaprobic | >3.0 | Heavy pollution |
| Site Photo |
Species 1 | Share of species 1 (%) | Index species 1 (s) | Species 2 | Share of species 2 (%) | Index species 2 (s) | Saprobity index (S) |
|---|---|---|---|---|---|---|---|
| Ilek 1 | Phragmites australis | 60 | 2.8 | Litter | 40 | 3.8 | 3.2 |
| Ilek 2 | Phragmites australis | 80 | 2.8 | – | 0 | – | 2.8 |
| Ilek 3 | Phragmites australis | 90 | 2.8 | – | 0 | – | 2.8 |
| Ilek 4 | Phragmites australis | 100 | 2.8 | – | 0 | – | 2.8 |
| Ilek 5 | Phragmites australis | 70 | 2.8 | Shrubs | 30 | 1.8 | 2.5 |
| Ilek 6 | Phragmites australis | 80 | 2.8 | Foam | 20 | 3.8 | 3.0 |
| Ilek 7 | Phragmites australis | 90 | 2.8 | – | 0 | – | 2.8 |
| Ilek 8 | Phragmites australis | 60 | 2.8 | Litter | 40 | 3.8 | 3.2 |
| Ilek 9 | Phragmites australis | 70 | 2.8 | Shrubs | 30 | 1.8 | 2.5 |
| Ilek 10 | Phragmites australis | 80 | 2.8 | – | 0 | – | 2.8 |
| Ilek 11 | Phragmites australis | 90 | 2.8 | – | 0 | – | 2.8 |
| Ilek 12 | Phragmites australis | 80 | 2.8 | Foam | 20 | 3.8 | 3.0 |
| Ilek 13 | Phragmites australis | 60 | 2.8 | Shrubs | 40 | 1.8 | 2.4 |
| Ilek 14 | Phragmites australis | 80 | 2.8 | Shrubs | 20 | 1.8 | 2.6 |
| Ilek 15 | Phragmites australis | 90 | 2.8 | – | 0 | – | 2.8 |
| Ilek 16 | Phragmites australis | 70 | 2.8 | Shrubs | 30 | 1.8 | 2.5 |
| Ilek 17 | Phragmites australis | 85 | 2.8 | – | 0 | – | 2.8 |
| Ilek 18 | Phragmites australis | 75 | 2.8 | Foam | 25 | 3.8 | 3.05 |
| Ilek 19 | Phragmites australis | 90 | 2.8 | – | 0 | – | 2.8 |
| Ilek 20 | Phragmites australis | 70 | 2.8 | Shrubs | 30 | 1.8 | 2.5 |
| Ilek 21 | Phragmites australis | 85 | 2.8 | – | 0 | – | 2.8 |
| Ilek 22 | Phragmites australis | 70 | 2.8 | Foam | 30 | 3.8 | 3.1 |
| Ilek 23 | Phragmites australis | 80 | 2.8 | – | 0 | – | 2.8 |
| Ilek well | Phragmites australis | 60 | 2.8 | Foam | 40 | 3.8 |
| Monitoring site | Saprobity index (S) | Visual signs | BOD₅ (mg/L) | COD (mg/L) | Petroleum hydrocarbons (mg/L) | Chromium Cr (mg/L) | Boron B (mg/L) | Other potential pollutants | Probable industrial source |
|---|---|---|---|---|---|---|---|---|---|
| Ilek 1 | 3.2 | litter, surface film | 6.2 | 34 | 0.32 | 0.018 | 0.28 | lead, nitrates | domestic sewage and illegal dumps |
| Ilek 6 | 3.0 | foam | 5.8 | 29 | 0.25 | 0.015 | 0.25 | copper, surfactants (SAS) | stormwater runoff |
| Ilek 8 | 3.2 | litter, surface film | 6.5 | 38 | 0.34 | 0.020 | 0.31 | phenols, ammonium | residential areas and car washes |
| Ilek 12 | 3.0 | foam | 5.7 | 32 | 0.28 | 0.017 | 0.26 | nitrites, zinc | livestock farms and agro-industrial enterprises |
| Ilek 18 | 3.05 | foam | 6.1 | 36 | 0.31 | 0.019 | 0.29 | ammonium, chlorides | agricultural fields and stormwater inflows |
| Ilek 22 | 3.1 | foam, greenish water | 6.4 | 39 | 0.35 | 0.021 | 0.33 | sulfates, phosphates | garden plots and suburban settlements |
| Ilek well | 3.2 | foam, surface film | 6.8 | 41 | 0.40 | 0.023 |
| Site № | BOD₅ (mg/L) | COD (mg/L) | Oil products (mg/L) | Cr (mg/L) | B (mg/L) | S (calculated) | Ecological assessment |
|---|---|---|---|---|---|---|---|
| 1 | 6.2 | 34 | 0.32 | 0.018 | 0.28 | 3.109 | Degradation |
| 6 | 5.8 | 29 | 0.25 | 0.015 | 0.25 | 3.00 | Relatively stable |
| 8 | 6.5 | 38 | 0.34 | 0.020 | 0.31 | 3.16 | Degradation |
| 12 | 5.7 | 32 | 0.28 | 0.017 | 0.26 | 3.02 | Degradation |
| 18 | 6.1 | 36 | 0.31 | 0.019 | 0.29 | 3.09 | Degradation |
| 22 | 6.4 | 29 | 0.35 | 0.021 | 0.33 | 3.16 | Degradation |
| 23 (well) | 6.8 | 41 | 0.40 | 0.023 | 0.35 | 3.26 | Degradation |
| Pollutant | Macrophyte(s) | Removal mechanism | Conditions of efficiency |
|---|---|---|---|
| Boron (B) | Lemna minor, Ceratophyllum demersum | Biosorption, complexation with polysaccharides | pH 6–8, high macrophyte density |
| Cr(VI) | Eichhornia crassipes, Typha latifolia | Reduction Cr(VI) → Cr(III), followed by sorption | Reducing environment, high biomass |
| Cr(VI) | Phragmites australis | Absorption, precipitation of Cr(III) in the rhizosphere | Presence of organics and reducing conditions |
| Boron (B) | Hydrilla verticillata | Involvement in plant metabolism, limited accumulation | Moderate concentrations (<1 mg/L) |
| Oil products | Eichhornia crassipes, Pistia stratiotes | Uptake, tissue accumulation, biodegradation | Temperature >20°C, absence of strong current |
| Surfactants | Lemna minor, Spirodela polyrhiza | Surface adsorption, biodegradation | Neutral pH, stable temperature |
| Ammonium (NH₄⁺) | Ceratophyllum demersum, Myriophyllum sp. | Uptake through plant surface, nitrification in rhizosphere | Low water flow, pH 6.5–7.5 |
| Nitrites/nitrates | Typha latifolia, Scirpus validus | Absorption and participation in denitrification | Moderate aeration, moderate flow |
| Phenols | Phragmites australis | Sorption, enzymatic degradation in rhizosphere | Presence of microaerobic conditions |
| Zinc (Zn), Copper | Eichhornia crassipes, Lemna gibba | Ion exchange, accumulation in leaves and roots | Concentrations up to 1 mg/L, pH 6–8 |
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