Article

Assessment of Water Quality and Ecological Integrity in an

Ecuadorian Andean Watershed
Freddy Armijos-Arcos 1 , Cristian Salazar 1,2, * , Andrés A. Beltrán-Dávalos 1 , Anna I. Kurbatova 2
and Elena V. Savenkova 2

1 Group of Research for Watershed Sustainabilty (GISOCH), Escuela Superior Politécnica de

Chimborazo (ESPOCH), Panamericana sur km 1 ½, Riobamba 060155, Ecuador
2 Department of Environmental Safety and Product Quality Management, Institute of Environmental
Engineering, Peoples’ Friendship University of Russia (RUDN University), 6 Miklukho-Maklaya Street,
117198 Moscow, Russia
* Correspondence: [email protected]

Abstract: This study assessed the water quality and ecological integrity of the Columbe
River micro-watershed in the Ecuadorian Andes through a multidimensional approach
incorporating biotic, physicochemical, and structural indices. Indices such as the Andean
Biotic Index (ABI), Biological Monitoring Working Party index adapted for Colombian
conditions (BMWP-Col), Fluvial Habitat Index (IHF), Riparian Quality Index adapted for
Andean conditions (QBR-And), and Water Quality Index (WQI) characterized environmen-
tal quality gradients and evaluated the impact of human activities across 11 monitoring
sites. Hierarchical cluster analysis classified sampling sites into three groups: less polluted
(LP), moderately polluted (MP), and highly polluted (HP). HP sites showed elevated levels
of biochemical oxygen demand (BOD5 ), chemical oxygen demand (COD), electrical conduc-
tivity (EC), and turbidity, alongside low biotic and structural scores, indicating advanced
ecological degradation. Conversely, LP sites demonstrated greater ecological integrity, de-
spite elevated BOD5 and COD levels across the watershed, suggesting widespread diffuse
contamination. The findings identify anthropogenic activities such as livestock, agricul-
ture, and domestic discharges as major pressures on water quality and macroinvertebrate
biodiversity. Significant correlations between physicochemical parameters—including
BOD5 and EC—and declining biotic indices underscore the link between chemical water
Academic Editor: Hossein Bonakdari
degradation and ecological fragmentation. In this context, this study highlights the critical
Received: 25 February 2025 need for comprehensive management and restoration strategies to combat pollution, safe-
Revised: 7 April 2025
guard relatively pristine areas, and rehabilitate the ecological integrity and connectivity of
Accepted: 16 April 2025
high-altitude Andean aquatic ecosystems under anthropogenic pressure.
Published: 18 April 2025

Citation: Armijos-Arcos, F.; Salazar,

Keywords: Andean watershed management; water quality assessment; macroinvertebrate
C.; Beltrán-Dávalos, A.A.; Kurbatova,
biodiversity; anthropogenic impact
A.I.; Savenkova, E.V. Assessment of
Water Quality and Ecological Integrity
in an Ecuadorian Andean Watershed.
Sustainability 2025, 17, 3684. https://
doi.org/10.3390/su17083684 1. Introduction
Copyright: © 2025 by the authors. The ecological integrity of fluvial ecosystems is essential for sustaining biodiversity,
Licensee MDPI, Basel, Switzerland. maintaining hydrological stability, and supporting water-dependent socio-economic ac-
This article is an open access article tivities. In mountainous regions such as the tropical Andes, these systems are particularly
distributed under the terms and
important due to their elevation-driven hydrological dynamics and ecological richness [1,2].
conditions of the Creative Commons
High-Andean micro-watersheds, in particular, support regional water security, but are
Attribution (CC BY) license
(https://creativecommons.org/
increasingly threatened by expanding agriculture, livestock grazing, deforestation, urban
licenses/by/4.0/). encroachment, and infrastructure development [2,3].

Sustainability 2025, 17, 3684 https://doi.org/10.3390/su17083684

Sustainability 2025, 17, 3684 2 of 32

The Andean landscape forms a complex mosaic shaped by both natural variability
and anthropogenic pressure. A steep topography and sharp altitudinal gradients pro-
duce sensitive hydrological systems, where páramos and riparian corridors help buffer
disturbances and maintain ecological balance [1,4]. However, the upward shift of land-use
activities—often replacing native vegetation with pastures or croplands—contributes to
erosion, nutrient loading, and habitat fragmentation [5,6]. In parallel, the limited reach
of wastewater treatment in rural areas results in untreated discharges, intensifying fe-
cal contamination and organic pollution [7,8]. These transformations reduce ecosystem
functionality, which becomes evident in degraded habitat structure and altered macroin-
vertebrate communities [2,3].
Ecological assessments based solely on physicochemical parameters offer only momen-
tary snapshots of water conditions and often fail to capture cumulative or chronic stressors.
For this reason, biotic indices like the Andean Biotic Index (ABI) and the Biological Mon-
itoring Working Party index adapted to Colombia (BMWP-Col) are widely used [9–12].
These tools evaluate macroinvertebrate community responses to pollution and help classify
ecosystem health based on species sensitivity to environmental stress [13,14]. Structural
indices such as the Fluvial Habitat Index (IHF) and the Riparian Forest Quality Index
adapted for Andean conditions (QBR-And) assess physical heterogeneity and riparian
condition, factors that are central to ecological resilience [12,15,16]. Additionally, the Water
Quality Index (WQI) integrates multiple physicochemical parameters to provide an overall
picture of water pollution, making it useful for characterizing pressure gradients in fluvial
systems [7,17].
Although these indices have proven useful in Andean contexts, many studies apply
them in isolation [2,7,10,11], which limits the understanding of how physical, chemical,
and biological factors interact. In several cases, tolerance values are based on generalized
regional datasets and may not reflect the specific environmental dynamics of local water-
sheds. Moreover, spatial methods that could reveal pollution gradients or priority areas for
conservation remain underused in routine biomonitoring.
A growing number of studies across the Andean region have begun to document
how environmental pressures are reflected in ecological indicators. For example, Hampel
et al. reported a sharp decline in riparian vegetation and fluvial habitat scores (QBR: from
82.4 to 10; IHF: from 60.3 to 39) in the Paute River Basin (Ecuador) due to cattle grazing
and riparian degradation [2]. Similarly, Vargas-Tierras et al. found a decrease in BMWP-
Col scores (from 72 to 47) in the Yanaquincha River linked to wastewater discharges [11].
Studies in Colombia, Peru, and Costa Rica have described comparable degradation patterns
caused by agricultural runoff, infrastructure development, and vegetation loss [12,18,19].
However, many of these works emphasize either biological or chemical metrics, rather than
using integrated frameworks.
This study addresses those methodological limitations by applying a multidimensional
approach to assess ecological integrity in the Columbe River micro-watershed (Ecuadorian
Andes). It combines biotic (ABI, BMWP-Col), structural (IHF, QBR-And), and physicochem-
ical (WQI) indices to produce a comprehensive evaluation of ecosystem health. In addition,
hierarchical clustering is used to classify sampling points spatially, uncovering patterns
of degradation and identifying zones with conservation or restoration potential. This
spatial classification not only supports the ecological diagnosis of the watershed but also
contributes to sustainable planning by identifying priority zones for targeted intervention.
By distinguishing areas requiring conservation from those demanding restoration, this
study provides actionable input for integrated watershed management. This aligns with
sustainability goals by promoting land-use decisions that maintain ecosystem services, re-
duce downstream contamination, and foster long-term resilience in socio-environmentally
Sustainability 2025, 17, 3684 3 of 32

vulnerable Andean regions. The application of this spatially explicit approach also supports
the local implementation of sustainability frameworks, such as payment for ecosystem
services (PES) and nature-based solutions, which have proven effective in reconciling
environmental conservation with rural development objectives [20,21].
The novelty of this research lies in the simultaneous application of five complementary
indices within a single Andean watershed, offering a more nuanced and accurate diagnosis
of ecological conditions. Unlike earlier studies that often rely on a limited set of indicators,
this approach enables site-specific interpretation and strengthens the operational basis for
watershed planning and intervention in data-scarce, environmentally pressured regions.

2. Materials and Methods

2.1. Study Area and Monitoring Samples
The present study was conducted in the Columbe River micro-watershed, which cov-
ers an area of 309.13 km2 and is located in the Columbe parish, Colta canton, Chimborazo
province. It features a characteristic Ecuadorian Andean climate, with an average annual
temperature of 11 ◦ C [22]. Its altitudinal range spans from 3080 to 4320 m above sea level,
contributing to remarkable microclimatic and ecological diversity [23]. Annual precipita-
tion varies significantly, with minimum values of 500 mm, maximum values of 1250 mm,
and an average of 780 mm [23]. The soil taxonomy in the Columbe parish is classified into
Inceptisols (44%), Mollisols (30%), Entisols (25%), and Histosols (1%), reflecting significant
edaphic heterogeneity [22]. Regarding land use and cover, agricultural areas dominate,
occupying 62.23% of the territory, followed by shrub and herbaceous vegetation (34.73%),
desertified areas (2.82%), anthropogenic zones (0.17%), and water bodies (0.04%) [22,23].
The ecosystems include Evergreen Montane Shrubland of the Northern Andes, Evergreen
Shrubland and Grassland of the Páramo, Páramo Grassland, Subnival Evergreen Grassland
and Shrubland of the Páramo, and Intervention Zones, reflecting the climatic and altitudinal
conditions of the micro-watershed [22,23]. The micro-watershed consists of páramo-origin
tributaries, including the Guashi River, Sasapud River, Guahuijón River, Llinllín Creek,
Pulucate Creek, Llinllín River, and several secondary drainages [24].
Sampling locations were strategically selected based on environmental representa-
tiveness. Priority was given to sites with direct effluent discharges, areas affected by
livestock and agricultural activities, and zones near human settlements to assess the impact
of anthropogenic activities. Additionally, locations in areas free from human settlements
were included to establish reference points for comparing impacted and non-impacted
environments. To ensure a representative sample, 11 monitoring points were selected
within the micro-watershed, as shown in Figure 1.
Point 1 was chosen in an area with no visible human disturbances, representing
minimally impacted headwaters. Points 2-to-11 were distributed along the watershed,
covering zones with varying degrees of agricultural use, livestock presence, and proximity
to human settlements.
Three monitoring sessions were conducted at each point to measure physicochemical
parameters, ensuring temporal consistency and data reliability. Macroinvertebrate sampling
and the corresponding components used to calculate ecological indices were assessed
through a single campaign. All field activities were carried out between September 2023
and March 2024 under comparable climatic conditions, ensuring uniform environmental
settings throughout the monitoring period.
Five indices were applied to assess water quality and ecological integrity in the
Columbe River micro-watershed, each addressing different ecological dimensions: aquatic
biodiversity (ABI, BMWP-Col), habitat structure (IHF, QBR-And), and physicochemical
status (WQI-TULSMA).
Sustainability 2025, 17, x FOR PEER REVIEW 4 of 33
Sustainability 2025, 17, 3684 4 of 32

Figure
Figure1.1.Geospatial
Geospatialdistribution
distributionof
ofsampling
samplingpoints
pointsin
inthe
theColumbe
ColumbeRiver
Rivermicro-watershed.
micro-watershed.

The indices
Five Andeanwere Bioticapplied
Index (ABI)
to assessandwater
the Biological
quality and Monitoring
ecological Working
integrityParty
in the in-
dex adapted
Columbe River for Colombia (BMWP-Col)
micro-watershed, each were chosen over
addressing other ecological
different alternatives,dimensions:
such as the
Neotropical
aquatic Lowland
biodiversity Stream
(ABI, Multimetric
BMWP-Col), Indexstructure
habitat (NLSMI)(IHF, or multimetric
QBR-And),indices like the
and physico-
Index of status
chemical Biotic Integrity (IBI) and the Multimetric Macroinvertebrate Index (MMI). These
(WQI-TULSMA).
alternatives
The Andean typically require
Biotic Indexgenus-
(ABI) or andspecies-level
the Biological taxonomic
Monitoring data,Working
which can be difficult
Party index
to obtainfor
adapted inColombia
regions with a limited were
(BMWP-Col) taxonomic
choseninfrastructure [25–27]. Moreover,
over other alternatives, such as theindices
Neo-
like the IBI and MMI may be less suitable for high-altitude environments,
tropical Lowland Stream Multimetric Index (NLSMI) or multimetric indices like the Index where the taxa
composition
of and environmental
Biotic Integrity dynamics differ
(IBI) and the Multimetric significantly from
Macroinvertebrate Index lowland
(MMI). systems
These[26,27].
alter-
The NLSMI, for example, was found to be less appropriate for
natives typically require genus- or species-level taxonomic data, which can be difficult the Guayas River Basin com-
to
pared to
obtain in the BMWP-Col
regions with a due to itstaxonomic
limited complexityinfrastructure
and the need [25–27].
for a wider range of indices
Moreover, metrics like
[25].
In contrast,
the IBI and MMI the ABImayand BMWP-Col
be less suitable rely on family level
for high-altitude identification
environments, and assign
where the taxaexplicit
com-
sensitivity
position and scores, making them
environmental more practical
dynamics and interpretable
differ significantly for applied
from lowland monitoring
systems [26,27]. in
high-Andean watersheds.
The NLSMI, for example, was found to be less appropriate for the Guayas River Basin
For structural
compared evaluation,
to the BMWP-Col due thetoFluvial Habitat Index
its complexity and the(IHF)need andforthe Riparian
a wider Vegetation
range of met-
Quality Index adapted for Andean conditions (QBR-And) were
rics [25]. In contrast, the ABI and BMWP-Col rely on family level identification and assign specifically chosen over
global tools
explicit like the
sensitivity Rivermaking
scores, Habitatthem Survey (RHS)
more or Rapid
practical andBioassessment
interpretable for Protocols
applied(RBPs).
mon-
These latter tools were primarily
itoring in high-Andean watersheds. developed for lowland temperate systems and are not as
effective in high-altitude
For structural Andean
evaluation, theenvironments
Fluvial Habitat dueIndex
to differences
(IHF) and in geomorphological
the Riparian Vegeta- and
ecological characteristics [28]. The RHS and RBPs are tailored
tion Quality Index adapted for Andean conditions (QBR-And) were specifically chosen to regions with more uniform
topography
over global toolsand less
like pronounced
the River Habitat altitude-driven
Survey (RHS) environmental gradients, making
or Rapid Bioassessment them
Protocols
less suitable for the heterogeneous and ecologically diverse
(RBPs). These latter tools were primarily developed for lowland temperate systems andnature of Andean rivers [28,29].
In contrast,
are although
not as effective inthe IHF was notAndean
high-altitude originally designed fordue
environments Andean systems, itinhas
to differences proven
geomor-
effective in capturing habitat features such as substrate diversity
phological and ecological characteristics [28]. The RHS and RBPs are tailored to regions and flow variability in
mountain
with more streams,
uniform makingtopographyit suitable
and lessfor pronounced
use in high-Andean environments
altitude-driven when adapted
environmental gra-
along with the QBR-And [2,12,19].
dients, making them less suitable for the heterogeneous and ecologically diverse nature
of Andean rivers [28,29]. In contrast, although the IHF was not originally designed for
Sustainability 2025, 17, 3684 5 of 32

The Water Quality Index (WQI) was chosen due to its alignment with national envi-
ronmental standards and its incorporation of region-specific parameters, making it a more
contextually relevant tool for assessing water quality in high-Andean ecosystems. In con-
trast to general models such as the NSF-WQI or CCME-WQI, which may not fully capture
local environmental variations, the WQI provides a tailored evaluation that reflects the
unique ecological and regulatory conditions of the region. This approach is consistent with
the findings of studies like Lukhabi et al. (2023) and Uddin et al. (2018), which emphasized
the importance of using locally adapted indices to enhance the accuracy of water quality
assessments, particularly in areas with distinct ecological characteristics [30,31].

2.2. Water Quality Parameters

Water samples were collected following the Standard Methods for the Examination
of Water and Wastewater (APHA, 2017). In situ physicochemical parameters—including
temperature, pH, electrical conductivity (EC), dissolved oxygen (DO), total dissolved solids
(TDS), and salinity—were measured using a Hanna Instruments HI98194 multiparameter
meter (Hanna Instruments, Woonsocket, RI, USA). This device features GLP (Good Lab-
oratory Practice) functions for calibration traceability and complies with IP67 standards,
ensuring durability under harsh field conditions [32].
For laboratory analyses, water samples were collected in appropriate containers and
preserved following standard protocols. Chemical parameters included sulfates (SO4 2− ),
analyzed by turbidimetry using a Hach TL2300 benchtop turbidimeter (Hach Company,
Loveland, CO, USA) (APHA 4500-SO4 2− E); phosphates (PO4 3− ), nitrates (NO3 − ), nitrites
(NO2 − ), and fluorides (F− ) were quantified using a Hach DR6000 UV–Vis spectrophotome-
ter (Hach Company, Loveland, CO, USA), applying the ammonium molybdate method
(APHA 4500-P E), UV spectrophotometry (APHA 4500-NO3 − B), Griess reagent method
(APHA 4500-NO2 − B), and SPADNS reagent method (APHA 4500-F− D), respectively.
Additionally, organic pollution indicators were assessed, including chemical oxy-
gen demand (COD), determined by the closed reflux method with potassium dichromate
using a Hach DRB200 digestion block (Hach Company, Loveland, CO, USA), followed
by spectrophotometric reading with a Hach DR6000 UV–Vis spectrophotometer (Hach
Company, Loveland, CO, USA) (APHA 5220 D), and biochemical oxygen demand over five
days (BOD5 ), measured by controlled incubation in a Memmert IN30 incubator (Memmert
GmbH, Schwabach, Germany) (APHA 5210 B). For microbiological parameters, fecal col-
iforms were quantified using a Millipore manifold filtration system, followed by incubation
on selective medium (m-FC agar) using the Memmert IN30 incubator (Memmert GmbH,
Schwabach, Germany) (APHA 9222 D).

2.3. Hydrological Characterization

Flow data for each sampling point were obtained from Valladares-Pogo (2024) [33],
using the velocity–area method recommended by the ICC [34]. Measurements involved di-
viding the river cross-section into segments, calculating area based on depth and width, and
recording water velocity using a Global Water FP111 flow meter (Global Water Instrumen-
tation, Gold River, CA, USA). The average velocity was estimated using the 0.2–0.8 depth
method, and a correction factor was applied based on stream characteristics, following
the recommendations outlined in the Hydrometry Manual developed by national author-
ities [35]. These measurements were taken during the same season as the water quality
assessments conducted for this study. The resulting values are shown in Table 1.
Sustainability 2025, 17, 3684 6 of 32

Table 1. Flow values at each sampling point. Reprinted with permission from ref. [33]. Copyright
2024 Escuela Superior Politécnica de Chimborazo.

Monitoring Point Flow (m3 /s)

P1 0.017
P2 0.623
P3 0.845
P4 0.379
P5 0.685
P6 0.419
P7 0.065
P8 0.156
P9 0.217
P10 0.166
P11 0.856

2.4. Sampling of Macroinvertebrates

Benthic macroinvertebrates were collected using a D-frame net and the kick sampling
method, a standardized procedure widely applied in bioassessment studies [36,37]. The net,
featuring a 30 cm-wide steel frame, a “D”-shaped opening, and a 500-micrometer mesh, is
designed to capture macroinvertebrates of various sizes while retaining fine particles. It was
positioned on the riverbed with the opening facing downstream. The substrate in front of
the net was disturbed by controlled foot kicks, dislodging organisms attached to sediments,
rocks, and organic matter. Sampling at each site was conducted for a standardized duration
of 3 min, ensuring thorough coverage of the main substrate types present (gravel, sand,
rocks, and aquatic vegetation).
Collected samples were transferred to pre-labeled containers with 70% alcohol for
preservation and transport to the laboratory. Macroinvertebrates were identified under a
Leica EZ4 stereoscope following the simplified protocol for evaluating the environmental
quality of Andean rivers (CERA-S) [16].

2.5. Determination of Biotic and Abiotic Indices

2.5.1. Aquatic Biotic Biodiversity
The ABI is based on the original BMWP and adapted for rivers located 2000 m above
sea level [13]. It considers the reduced number and distribution of macroinvertebrate
families in high-elevation ecosystems, where altitude limits both diversity and sensitivity
to disturbance [9,13]. Families are assigned tolerance scores from 1 (high tolerance) to 10
(high sensitivity), and the index value is calculated as the sum of scores for all families
present at each site. Sites are classified into five ecological quality levels: >96 (very good),
59–96 (good), 35–58 (moderate), 14–34 (poor), and <14 (very poor) [9].
The BMWP-Col is an adaptation of the original BMWP to Colombian high-mountain
rivers and is widely used in Andean ecosystems [9,14]. Its development was based on an
exhaustive review of water quality studies using macroinvertebrates, allowing for contex-
tualization and adaptation to the specific taxonomic and environmental conditions of the
Andean region [14]. The index considers only the presence or absence of macroinvertebrate
families, each assigned a tolerance score ranging from 1 (tolerant) to 10 (sensitive). The
final score for each site corresponds to the sum of the values of all families present. The
classification categories are as follows: >150 (very-clean-to-clean water), 61–100 (slightly
polluted), 36–60 (moderately polluted), 16–35 (heavily polluted), and <15 (very heavily
polluted) [14].
While both indices are based on macroinvertebrate sensitivity to pollution, the ABI is
specifically tailored to high-altitude Andean rivers with reduced faunal richness, whereas
Sustainability 2025, 17, 3684 7 of 32

the BMWP-Col encompasses a broader range of families, representative of the ecological

variability across Andean mountain systems.

2.5.2. Fluvial Habitat Structure

The Fluvial Habitat Index (IHF) evaluates the physical quality of riverine environments
based on seven specific components: substrate inclusion, frequency of riffles, substrate
composition, flow velocity and depth regimes, riverbed shading, structural heterogeneity,
and aquatic vegetation cover [15]. Each component is scored independently, contributing
to a maximum possible score of 100. Higher values indicate more diverse and ecologically
functional river habitats. The ecological condition is categorized as follows: ≥90 (very high
diversity), 71–80 (high), 50–70 (moderate), 31–49 (low), and 0–30 (very low) [15]. Table 2
summarizes the components, scoring rationale, and ecological interpretation.

Table 2. Description of the components included in the Fluvial Habitat Index (IHF). Adapted from
Pardo et al. (2002) [15].

IHF Component Definition Ecological Relevance Maximum Score

Refers to the presence of Less embeddedness supports
Substrate inclusion and
compacted sand between more habitat availability for 10
limitation
coarse substrates. macroinvertebrates.
Assesses the number of Riffles create habitat diversity
Frequency of riffles 10
riffles in the reach. and enhance oxygenation.
Evaluates the diversity of Diverse substrates support
Substrate composition 20
substrate types present. varied benthic communities.
Assesses the presence of A wider range of flow
Speed/depth regimes combinations of flow conditions promotes 10
velocity and depth. species richness.
Estimates shading from Shading regulates temperature
Shade on the riverbed 10
riparian vegetation. and supports aquatic life.
Considers the presence of
Structural complexity improves
Riverbed heterogeneity woody debris, roots, and 10
refuge and habitat quality.
natural barriers.
Quantifies aquatic Vegetation supports food webs
Aquatic vegetation cover 30
vegetation types. and habitats for colonization.

The QBR-And, proposed by Acosta et al. (2009) [16], is an adaptation of the Riparian
Vegetation Quality Index (QBR) developed by Munné et al. (2003) [38]. It is specifically
designed to evaluate the unique characteristics of Andean vegetation formations and
associated riparian typologies, such as rocky banks, páramos, and punas. The QBR-
And focuses on four components: vegetation cover, structural complexity, vegetation
composition (native vs. introduced), and the degree of channel naturalness. Each category
is scored on a tiered scale (0, 5, 10, or 25), and adjustment factors (ranging from −10 to
+10) may be applied according to the criteria in the CERA protocol, depending on aspects
such as forest connectivity or distribution pattern. The total score ranges from 0 to 100,
with quality classifications defined as ≥96 (very good), 76–95 (slightly altered), 51–75
(moderately altered), 26–50 (strongly altered), and ≤25 (extremely degraded) [16]. Table 3
outlines the component definitions and their ecological significance.
Sustainability 2025, 17, 3684 8 of 32

Table 3. Description of the components included in the adapted Riparian Vegetation Quality Index
for the Andes (QBR-And). Adapted from Acosta et al. (2009) [16].

QBR-and Component Definition Ecological Relevance Maximum Score

Greater cover implies better
Evaluates vegetation cover
Riparian zone coverage erosion control and 25
on riverbanks.
habitat quality.
Structural complexity is
Assesses vertical stratification
Vegetation structure associated with mature 25
and species diversity.
riparian systems.
Evaluates the presence of Native vegetation and minimal
Riparian vegetation quality native vs. exotic species and disturbance increase 25
anthropogenic impact. ecological value.
Natural channels offer better
Considers the extent of
Degree of naturalness habitat continuity and 25
channel modification.
ecological function.

2.5.3. Physicochemical and Microbiological Water Quality

Anthropogenic stress was assessed through the calculation of a Water Quality Index
(WQI), integrating physicochemical and microbiological parameters, with reference to
the maximum permissible limits to preserve aquatic life specified in Annex 1 of Book
VI of the Unified Text of Secondary Legislation of the Ministry of the Environment of
Ecuador (TULSMA, for its acronym in Spanish). The parameters considered for the WQI
calculation in this study are those that have established limits in the TULSMA regulations,
including true color, biochemical oxygen demand (BOD5 ), chemical oxygen demand,
fluorides, nitrates, nitrites, dissolved oxygen, pH, total dissolved solids, sulfates, turbidity,
and fecal coliforms.
The WQI calculation involved three steps [17]. The first step determined weights for
the analyzed parameters, ranging from 1 to 5. These weights reflect the relative importance
of each parameter in maintaining water quality for aquatic life. Using these weights, the
relative weight of each parameter was computed with Equation (1):

wi
Wi = (1)
∑in = 1 wi

where Wi is the relative weight, wi is the weight of each parameter, and n is the total number
of parameters.
The second step involved determining the quality rating for each parameter by divid-
ing its concentration in the water sample by the standard established in the Ecuadorian
TULSMA regulations and multiplying the result by 100 (Equation (2)).

Ci
qi = × 100 (2)
Si

where qi represents the quality rating, Ci is the parameter concentration, and Si is the
standard value from TULSMA.
Finally, the WQI evaluation determined a sub-index for each parameter and then
summed all sub-indices for each sample (Equations (3) and (4)):

SIi = Wi × qi (3)

n
WQI = ∑ SIi (4)
i=1
Sustainability 2025, 17, 3684 9 of 32

where SIi is the sub-index for the i-th parameter.

Based on the final WQI value, water quality is classified into five categories: 0–25 (very
clean), 26–50 (clean), 51–75 (moderately polluted), 76–100 (polluted), and >100 (dirty) [17].
A unified scale was used to compare methods for parametrizing ecological integrity
and water quality in the Andean micro-watershed. The threshold values for the ranges
of the ABI, BMWP-Col, IHF, QBR-And, and WQI indices were reviewed, resulting in the
identification of five classes, as shown in Table 4.

Table 4. Classifications and scores of used indices.

Classification
Excellent Good Fair Poor Very Poor
Index
Biotic indices
ABI >96 59–96 35–58 14–34 <14
BMWP-Col ≥150 61–100 36–60 16–35 <15
Abiotic indices
IHF ≥90 71–80 50–70 31–49 0–30
QBR-And ≥96 76–95 51–75 26–50 ≤25
Physicochemical index
WQI ≤25 26–50 51–75 76–100 >100

2.6. Data Processing

The drainage basin of the Columbe River was delineated using a 10 m-resolution
Digital Elevation Model (DEM) and processed in QGIS 3.16. Hydrological tools, including
fill sinks, flow direction, and flow accumulation, were applied to generate the watershed
boundary and stream network. This geospatial delineation was verified against official
topographic maps provided by the Military Geographic Institute of Ecuador (IGM, for its
acronym in Spanish) [39], ensuring consistency with national cartographic standards.
The Pearson correlation coefficient was calculated to identify significant relationships
(p < 0.01) between the water quality parameters and the ecological indices.
To explore spatial patterns, a hierarchical agglomerative cluster analysis (CA) was
performed using the Ward linkage method and Euclidean distance applied to standardized
values (Z-scores) of 22 variables: 17 physicochemical parameters and 5 ecological indices.
Standardization was necessary to homogenize variable scales and ensure comparability
across different units of measurement. This clustering method was selected due to its
demonstrated ability to produce compact and interpretable groupings, which is particularly
advantageous in environmental assessments [40,41]. To validate the robustness of the
clustering solution, a sensitivity analysis was carried out by comparing results across
alternative linkage methods (single, complete, average) and distance metrics (cityblock
and correlation). Furthermore, the agglomeration coefficient schedule was reviewed to
determine the optimal number of clusters. A marked increase in the coefficient value
between successive fusion stages was used as the criterion for selecting the cut-off point,
thereby avoiding reliance solely on visual inspection of the dendrogram.
To assess whether the clusters significantly differed in terms of environmental con-
ditions, a one-way ANOVA was applied to each of the 22 variables. When statistically
significant differences were found (p < 0.05), Tukey’s Honestly Significant Difference (HSD)
post hoc test was used to determine which specific groups differed from each other.
All statistical analyses were conducted using RStudio (version 2023.12.0).
Sustainability 2025, 17, 3684 10 of 32

3. Results
3.1. Aquatic Biotic Biodiversity
A total of 19 benthic macroinvertebrate families were identified across the 11 mon-
itoring points (Table 5). Points 3 and 8 registered the highest richness, with 11 families
each, including sensitive taxa such as Baetidae, Elmidae, and Perlidae. In contrast, points 6
and 5 showed the lowest richness, with four and five families, respectively, where tolerant
taxa such as Elmidae and Oligochaeta predominated. Other points exhibited intermediate
richness (six-to-nine families), with a mix of tolerant and sensitive groups.

Table 5. Summary of benthic macroinvertebrate families in the sampling sites.

Families P1 P2 P3 P4 P5 P6 P7 P8 P9 P10 P11

Baetidae x x x x x x
Blephariceridae
Chironomidae x x x x
Elmidae x x x x x x x x x
Glossiphoniidae x x
Hyalellidae x x x x x x
Hydracarina x
Hydrobiosidae x x x x x
Leptoceridae x x
Limnephilidae x x
Lymnaeidae
Odontoceridae x x
Oligochaeta x x x x x x x x x x
Perlidae x x x x x x x
Scirtidae
Simuliidae x x x x x x x x
Sphaeriidae x x x
Tabanidae x
Turbellaria x x x x

The identified families were used to compute the BMWP-Col and ABI indices.
Tables 6 and 7 present the family level scores used for each index, applied at every sam-
pling point. Both indices assign scores based on pollution tolerance: lower values denote
tolerance, while higher values reflect sensitivity to environmental disturbance.

Table 6. BMWP-Col scores assigned to macroinvertebrate families at each sampling point.

BMWP-Col Scores by Sampling Point

Family
P1 P2 P3 P4 P5 P6 P7 P8 P9 P10 P11
Baetidae 7 - 7 7 - - 7 7 - - 7
Blephariceridae 10 10 10 10
Chironomidae - 2 2 2 - - - 2 - - -
Elmidae 6 6 6 - 6 6 - 6 6 6 6
Glossiphoniidae 7 - 7 - - - - - - - -
Hyalellidae 7 7 - - - - - 7 7 7 7
Hydrobiosidae - - 9 - - 9 9 - 9 - 9
Leptoceridae - - - - 8 - 8 - - - -
Limnephilidae - - - 7 - - - - - - 7
Lymnaeidae - - - 4 5 - - 4 4 4 4
Odontoceridae 10 10 - - - - - - - - -
Perlidae 10 10 10 10 - 10 - 10 - 10 -
Scirtidae 7 - - - - - - 7 - - -
Simuliidae - 8 8 8 8 8 8 8 - 8 -
Sphaeriidae - - - 4 - - - - - 4 4
Tabanidae 5
Turbellaria 7 - 7 - - - - - - 7 7
Sustainability 2025, 17, 3684 11 of 32

Table 7. ABI scores assigned to macroinvertebrate families at each sampling point.

ABI Scores by Sampling Point

Family
P1 P2 P3 P4 P5 P6 P7 P8 P9 P10 P11
Baetidae 4 - 4 4 - - 4 4 - - 4
Chironomidae - 2 2 2 - - - 2 - - -
SustainabilityElmidae
2025, 17, x FOR PEER REVIEW
5 5 5 - 5 5 - 5 5 5 11 of 533
Glossiphoniidae 6 - 6 - - - - - - - -
Hyalellidae 6 6 - - - - - 6 6 6 6
Hydracarina - - - - - - - 4 - - -
Hydracarina - - - - - - - 4 - - -
Hydrobiosidae - - 8 - - 8 8 - 8 - 8
Hydrobiosidae
Leptoceridae - - - - 8 - - - - 8 8 - 8 8 - - 8 - - - 8 -
Leptoceridae
Limnephilidae - - - - - - - 7 8 - - - 8 - - - - - - - - 7
Limnephilidae
Odontoceridae - 10 - 10 - - 7 - - - - - - - - - - - - - 7 -
Odontoceridae
Oligochaeta 10 1 10 1 - 1 - 1 - 1 - - - 1 - 1 - 1 - 1 - 1
Oligochaeta
Perlidae 1 10 1 10 1 10 1 10 1 - - 10 1 - 1 10 1 - 1 10 1 -
Simuliidae
Perlidae 10 - 10 5 10 5 10 5 - 5 10 5 - 5 10 5 - - 10 5 - -
Sphaeriidae
Simuliidae - - 5 - 5 - 5 3 5 - 5 - 5 - 5 - - - 5 3 - 3
Tabanidae
Sphaeriidae - - - 4 - - 3 - - - - - - - - - - - 3 - 3 -
Turbellaria
Tabanidae - 5 4 - - 5 - - - - - - - - - - - - - 5 - 5
Turbellaria 5 - 5 - - - - - - 5 5

According to Table 6 (BMWP-Col index), Perlidae and Odontoceridae consistently

According to Table 6 (BMWP-Col index), Perlidae and Odontoceridae consistently
received the highest scores (10), reflecting their sensitivity to pollution and association
received the highest scores (10), reflecting their sensitivity to pollution and association
with cleaner conditions. Elmidae and Simuliidae were also frequently recorded across
with cleaner conditions. Elmidae and Simuliidae were also frequently recorded across
sites, often receiving moderate scores, indicating that they can tolerate a broader range of
sites, often receiving moderate scores, indicating that they can tolerate a broader range of
environmental conditions [14]. Chironomidae, a tolerant group, received low scores (2),
environmental conditions [14]. Chironomidae, a tolerant group, received low scores (2),
suggesting their presence in more disturbed environments [14].
suggesting their presence in more disturbed environments [14].
In Table
In Table 77 (ABI),
(ABI),similar
similartrends
trendswere
wereobserved.
observed.Families
Families such
such as as Oligochaeta
Oligochaeta andand Chi-
Chi-
ronomidae received low scores (1–2), consistent with sites that exhibited poorer
ronomidae received low scores (1–2), consistent with sites that exhibited poorer ecological ecological
conditions [13]. On
conditions [13]. Onthe
theother
otherhand,
hand,Perlidae,
Perlidae,Odontoceridae,
Odontoceridae, and
and Hydrobiosidae
Hydrobiosidae received
received
higher scores (8–10), indicating that they are found in environments with better
higher scores (8–10), indicating that they are found in environments with better ecological ecological
quality and lower
quality and lower pollution
pollutionlevels
levels[13].
[13].
Figure 2 shows the total index
Figure 2 shows the total index valuesvaluescalculated
calculatedfor
foreach
eachsampling
sampling point
point using
using thethe
scores from Tables
scores from Tables 66 and
and 7.
7.

Figure 2. Calculated
Calculatedbiotic
bioticindices
indicesvalues
values(BMWP
(BMWPand
andABI).
ABI).

The BMWP-Col recorded high values, indicating fair-to-good water quality at points
1, 2, 3, 4, 7, 8, 10, and 11, with point 3 showing the highest value (71). Low values, reflecting
poor water quality, were observed at points 5, 6, and 9, with point 9 being the lowest (26).
The ABI showed a similar trend, with high values (fair water quality) at points 1, 2, 3, 8,
Sustainability 2025, 17, 3684 12 of 32

The BMWP-Col recorded high values, indicating fair-to-good water quality at points 1,
2, 3, 4, 7, 8, 10, and 11, with point 3 showing the highest value (71). Low values, reflecting
poor water quality, were observed at points 5, 6, and 9, with point 9 being the lowest (26).
The ABI showed a similar trend, with high values (fair water quality) at points 1, 2, 3, 8,
10, and 11. Point 3 again recorded the highest value (50). Poor water quality was evident
Sustainability 2025, 17, x FOR PEER REVIEW 12 ofby
at points 4, 5, 6, 7, and 9, with point 5 showing the lowest value (19), closely followed 33

point 9 (20).

3.2.
3.2. Fluvial
Fluvial Habitat
Habitat Structure
Structure
Based on the component
Based on the component scores detailed
scores in Tables
detailed 2 and23,and
in Tables the 3,
final
thevalues of the Fluvial
final values of the
Habitat Index (IHF) and the Riparian Forest Quality Index adapted for Andean
Fluvial Habitat Index (IHF) and the Riparian Forest Quality Index adapted for Andean conditions
(QBR-And) are presented
conditions (QBR-And) areinpresented
Figure 3. in Figure 3.

Figure 3. Values
Figure3. Valuesof
ofhabitat
habitatstructure
structureindices
indices(IHF
(IHFand
andQBR-AND).
QBR-AND).

The
TheFluvial
FluvialHabitat
HabitatIndex
Index(IHF)
(IHF)classified
classifiedhabitat
habitatquality
qualityasas “good”
“good” at at
points
points1, 1,
2, 3,
2, 5,
3,
and 6, with
5, and the the
6, with highest values
highest recorded
values at points
recorded 2, 3, and
at points 2, 3,6and
(73).6 Habitat qualityquality
(73). Habitat was “fair”
was
at points
“fair” at 4, 7, 8, 9,
points 4, 10, and
7, 8, 9, 11,
10, with thewith
and 11, lowestthevalues
lowestobserved at points at
values observed 9 and 10 (58).
points 9 and 10
(58).The Riparian Vegetation Quality Index adapted for Andean conditions (QBR-And)
classified habitat quality
The Riparian as “good”
Vegetation at points
Quality Index1,adapted
2, and 6,forwith point conditions
Andean 2 showing the highest
(QBR-And)
value (90). Habitat quality was “fair” at points 3, 5, 7, 8, 9, and 11 and “poor”
classified habitat quality as “good” at points 1, 2, and 6, with point 2 showing the highest at points 4
and
value10,(90).
withHabitat
point 4quality
recordingwasthe lowest
“fair” value (40).
at points 3, 5, 7, 8, 9, and 11 and “poor” at points 4
and 10, with point 4 recording the lowest value (40).
3.3. Physicochemical and Microbiological Water Quality
The averages ofand
3.3. Physicochemical theMicrobiological
values obtained from
Water the sampling conducted at the 11 study
Quality
points are presented in Table 8.
The averages of the values obtained from the sampling conducted at the 11 study
Apparent color values ranged from 35 CU at P3 to 230 CU at P10. Although TULSMA
points are presented in Table 8.
does not specify a standard for apparent color, elevated levels at P9 (192.5 CU), P10 (230
CU), and P11 (210 CU) suggest a high concentration of suspended and dissolved substances.
Table 8. Mean value of physical, chemical, and biological parameter analysis.
True color values, on the other hand, remained below the TULSMA threshold of 100 PCU,
with the TULSMA
Parameter (Measure Unit) P1 P2 highest
P3 value
P4 recorded
P5 P6at P7 (80
P7 PCU).P8Electrical
P9 conductivity
P10 (EC)
P11 exhibited the
Standard
highest readings at P10 (473.8 µS/cm) and P9 (423.5 µS/cm), indicating elevated ionic
Apparent color (CU) 40 52.50 35 65 83.75 47.19 63.98 97.50 192.50 230 210 -
content. Salinity levels remained low, ranging from 0.05 ppt at P2 to 0.33 ppt at P4, with no
True color (PCU) 30 45 55 55 65 65 80 65 20 20 30 100
EC (uS/cm) 128.6 98.4 125.6 123.3 119.4 118.9 110.5 166.7 423.5 473.8 387.8 -
BOD5 (mg/L) 27.8 49.8 28.3 33.5 27.3 23.6 40.3 30.6 30 30.5 31.8 20
COD (mg/L) 55.6 99.6 56.6 67 54.6 47.3 80.6 61.3 60 61 63.6 40
Fluorides (mg/L) 0.29 0.22 0.28 0.53 0.40 0.40 0.39 0.23 1.07 0.93 0.89 1.60
Phosphates (mg/L) 2.56 0.67 0.85 1.68 0.82 0.93 2.69 1.07 3.12 3.03 2.46 -
Sustainability 2025, 17, 3684 13 of 32

TULSMA standard for direct comparison. Total dissolved solids (TDS) were also within
permissible limits, peaking at 237.3 mg/L at P10, well below the 500 mg/L threshold.
Temperature readings ranged from 10.5 ◦ C at P2 to 16.4 ◦ C at P10. Turbidity presented
notable exceedances at P9 (18.7 NTU), P10 (20.3 NTU), and P11 (17 NTU), surpassing the
10 NTU limit set by TULSMA.

Table 8. Mean value of physical, chemical, and biological parameter analysis.

TULSMA
Parameter (Measure Unit) P1 P2 P3 P4 P5 P6 P7 P8 P9 P10 P11
Standard
Apparent color (CU) 40 52.50 35 65 83.75 47.19 63.98 97.50 192.50 230 210 -
True color (PCU) 30 45 55 55 65 65 80 65 20 20 30 100
EC (uS/cm) 128.6 98.4 125.6 123.3 119.4 118.9 110.5 166.7 423.5 473.8 387.8 -
BOD5 (mg/L) 27.8 49.8 28.3 33.5 27.3 23.6 40.3 30.6 30 30.5 31.8 20
COD (mg/L) 55.6 99.6 56.6 67 54.6 47.3 80.6 61.3 60 61 63.6 40
Fluorides (mg/L) 0.29 0.22 0.28 0.53 0.40 0.40 0.39 0.23 1.07 0.93 0.89 1.60
Phosphates (mg/L) 2.56 0.67 0.85 1.68 0.82 0.93 2.69 1.07 3.12 3.03 2.46 -
Nitrates (mg/L) 0.25 0.30 0.40 0.40 0.35 0.26 0.27 1.05 0.55 4.65 0.50 5
Nitrites (mg/L) 0.006 0.006 0.005 0.005 0.005 0.005 0.005 0.004 0.009 0.011 0.006 0.05
Dissolved oxygen (%sat) 59.4 58.9 59.4 56.8 57.7 58.4 57.7 62.2 58.2 60.3 60.3 80
pH 7.63 7.38 7.51 7.49 7.47 7.49 7.50 7.62 7.63 7.67 7.72 6–9
Salinity (ppt) 0.06 0.05 0.06 0.33 0.15 0.17 0.21 0.08 0.22 0.23 0.19 -
TDS (mg/L) 64.4 49.3 62.9 61.8 60 60 55.5 82.8 218.3 237.3 197 500
Sulphates (mg/L) 0 0 0 0 0 0 0 0 10.5 10 7.5 250
Temperature (◦ C) 10.7 10.5 12.4 11.8 12.2 11.9 11.1 13.8 15.6 16.4 16.2 -
Turbidity (NTU) 2.3 2.9 2.6 3.8 9.7 3.1 3.1 5.6 18.7 20.3 17 10
Fecal coliforms
0 0 0 10 0 0 10 20 140 20 30 200
(CFU/100 mL)

Biochemical oxygen demand (BOD5) levels exceeded the TULSMA limit of 20 mg/L at
all sampling points, with the highest concentration at P2 (49.8 mg/L), reflecting significant
organic pollution across the watershed. Similarly, chemical oxygen demand (COD) levels
were elevated, ranging from 47.3 mg/L at P6 to 99.6 mg/L at P2, surpassing the maximum
permissible value of 40 mg/L, further indicating substantial organic and inorganic pollutant
loads. Fluoride concentrations ranged from 0.22 mg/L at P2 to 1.07 mg/L at P9, remaining
within the TULSMA limit of 1.60 mg/L. Phosphate concentrations peaked at P9 (3.12 mg/L)
and P10 (3.03 mg/L), though no TULSMA standard is specified for this parameter. Nitrate
concentrations were consistently below the 5 mg/L limit, with a maximum value of 4.65
mg/L at P10. Nitrite levels were low across all sites, with a maximum of 0.011 mg/L at
P10, well within the 0.05 mg/L threshold. Dissolved oxygen (DO) levels were critically low,
ranging from 56.8% saturation at P4 to 62.2% at P8, failing to meet the TULSMA standard
of 80% saturation. Such hypoxic conditions pose a risk to aquatic biota and reflect elevated
organic matter decomposition. pH values remained stable, between 7.38 at P2 and 7.72 at
P11, complying with the acceptable range of 6–9 established by TULSMA. Sulphates were
detected only at P9 (10.5 mg/L), P10 (10 mg/L), and P11 (7.5 mg/L), significantly lower
than the 250 mg/L limit.
Fecal coliform concentrations remained below the TULSMA limit of 200 CFU/100 mL
at all sampling points. The highest concentrations were observed at P9 (140 CFU/100 mL),
P10 (20 CFU/100 mL), and P11 (30 CFU/100 mL). Notably, no fecal coliforms were detected
at P1, P2, P3, P5, and P6.
Figure 4 presents the WQI values obtained for the 11 monitoring points along the
Columbe River micro-watershed.
Sustainability 2025, 17, 3684
Sustainability 2025, 17, x FOR PEER REVIEW 1414of
of 33
32

Sustainability 2025, 17, x FOR PEER REVIEW 14 of 33

4. Values of WQI TULSMA.

Figure 4.
Figure

AccordingAnalysis
3.4. Correlation to the Water Quality
and Spatial Index values,
Classification water qualityQuality
of Environmental was classified as “good”
at points 1, 3, and 6, with values ranging from 46 to 49. In contrast, water quality was
3.4.1. Correlation Patterns Among Physicochemical and Biological Variables
classified as “fair” at points 2, 4, 5, 7, 8, 9, and 11, with values ranging from 54 to 73, while
Figure
point Figure
10 5 presents
4. Values
showed ofaWQI a heatmap
TULSMA.
value of Pearson correlation
of 79, corresponding coefficients,
to “poor” water with statistically sig-
quality.
nificant correlations (p < 0.01) highlighted by circles.
3.4. Correlation Analysis and Spatial Classification of Environmental Quality
3.4.1. Correlation
3.4.1. Correlation Patterns
Patterns Among
Among Physicochemical
Physicochemical and and Biological
Biological Variables
Variables
Figure 55 presents
Figure presentsaaheatmap
heatmapofof Pearson
Pearson correlation
correlation coefficients,
coefficients, withwith statistically
statistically sig-
significant correlations (p < 0.01) highlighted by circles.
nificant correlations (p < 0.01) highlighted by circles.

Figure 5. Heatmap of Pearson correlation coefficients. The values highlighted with circles repre-
sent statistically significant correlations (p < 0.01).

Apparent color showed significant positive correlations with electrical conductivity,

fluorides, TDS, sulfates, and turbidity. Electrical conductivity was significantly correlated
with TDS, sulfates, and turbidity. BODcoefficients.
Figure 5. Heatmap of Pearson correlation coefficients. repre-
The values highlighted with circles represent
5 exhibited a significant correlation with COD. Flu-
statistically
sent significant
statistically correlations
significant (p < 0.01).
correlations (p < 0.01).
orides were positively correlated with sulfates, while TDS showed significant relation-
ships with sulfates and turbidity. The Fluvial Habitat Index (IHF) had statistically signif-
Apparent color showed significant positive correlations with electrical conductivity,
icant negative correlations of moderate-to-strong intensity (r between −0.81 and −0.88)
fluorides, TDS, sulfates, and turbidity. Electrical conductivity was significantly correlated
with TDS, sulfates, and turbidity. BOD5 exhibited a significant correlation with COD. Flu-
orides were positively correlated with sulfates, while TDS showed significant relation-
Sustainability 2025, 17, 3684 15 of 32

Apparent color showed significant positive correlations with electrical conductivity,

fluorides, TDS, sulfates, and turbidity. Electrical conductivity was significantly correlated
with TDS, sulfates, and turbidity. BOD5 exhibited a significant correlation with COD. Fluo-
rides were positively correlated with sulfates, while TDS showed significant relationships
Sustainability 2025, 17, x FOR PEER REVIEW 15 of 33
with sulfates and turbidity. The Fluvial Habitat Index (IHF) had statistically significant
negative correlations of moderate-to-strong intensity (r between −0.81 and −0.88) with
apparent
with color,color,
apparent electrical conductivity,
electrical fluorides,
conductivity, phosphates,
fluorides, phosphates,TDS, sulfates,
TDS, temperature,
sulfates, temper-
and turbidity. The Andean Biotic Index (ABI) demonstrated a strong, statistically
ature, and turbidity. The Andean Biotic Index (ABI) demonstrated a strong, statistically signifi-
cant positive
significant correlation
positive with the
correlation Biological
with Monitoring
the Biological WorkingWorking
Monitoring Party index adapted
Party indexfor
adapted for Colombia (BMWP-Col). The Water Quality Index (WQI) showed a moderatecor-
Colombia (BMWP-Col). The Water Quality Index (WQI) showed a moderate negative
negative = −0.77) with
relation (rcorrelation (𝑟 = the Fluvial
−0.77) withHabitat Index
the Fluvial (IHF) and
Habitat moderate
Index (IHF) andpositive correlations
moderate posi-
(r between 0.75 and 0.80) with apparent color, electrical conductivity, nitrites,
tive correlations (𝑟 between 0.75 and 0.80) with apparent color, electrical conductivity, TDS, sulfates,
and turbidity.
nitrites, TDS, sulfates, and turbidity.

3.4.2. Cluster
3.4.2. ClusterAnalysis
Analysisof ofPollution
PollutionGradients
Gradients
Sampling sites
Sampling sites were
were classified
classified based
based onon their
theirpollution
pollutionlevels
levelsusing
usinghierarchical
hierarchicalclus-
cluster
analysis that incorporated all evaluated physicochemical and microbiological
ter analysis that incorporated all evaluated physicochemical and microbiological param- parameters,
along along
eters, with the
withcalculated indices.
the calculated indices.
To assess
assess the
the consistency
consistencyofofthe theclustering
clusteringoutcome,
outcome, a sensitivity
a sensitivity analysis
analysis was was
con- con-
ducted by
ducted by reapplying
reapplyingthe thehierarchical
hierarchicalcluster
clusteranalysis
analysisusing
using alternative
alternative linkage
linkage methods
methods
(single, complete,
complete,and andaverage)
average)and anddistance
distancemetrics
metrics (cityblock
(cityblock andand correlation).
correlation). TheThe
com- com-
parison revealed that,
parison revealed that,although
althoughminor minorvariations
variationsoccurred
occurred inin the
the arrangement
arrangement of of specific
specific
sites, the
sites, theoverall
overallstructure
structure remained
remained stable
stable across
across methods.
methods. This This consistency
consistency reinforces
reinforces the
methodological
the methodological reliability of the
reliability chosen
of the chosenconfiguration—Ward’s
configuration—Ward’s linkage
linkageandandEuclidean
Euclidean
distance—for
distance—for classifying
classifyingecological
ecologicalconditions
conditionsininthe thestudy
studyarea.
area.
Additionally,
Additionally,the theagglomeration
agglomeration coefficient
coefficientcurve waswas
curve analyzed to determine
analyzed the op-the
to determine
timal
optimal number
numberof clusters. A marked
of clusters. A marked increase in theincoefficient
increase was observed
the coefficient was observedbetween fu-
between
sion stages 8 and 9 (from 5.91 to 10.00), indicating that merging beyond
fusion stages 8 and 9 (from 5.91 to 10.00), indicating that merging beyond this point wouldthis point would
combine
combine ecologically
ecologicallydistinct
distinctgroups.
groups.This Thisinflection
inflectionpoint
pointsupported
supported thetheselection of of
selection a a
three-cluster solution as the most representative configuration of the dataset
three-cluster solution as the most representative configuration of the dataset (Figure 6). (Figure 6).

Figure 6.
Figure Dendrogramillustrating
6. Dendrogram illustratingspatial
spatialclustering
clustering
ofof sampling
sampling sites
sites based
based onon water
water quality
quality andand
ecological indices.
ecological indices.LP:
LP:less
lesspolluted,
polluted,MP:
MP:moderately
moderatelypolluted,
polluted, HP:
HP: highly
highly polluted.
polluted.

Based on the dendrogram (Figure 6), three distinct clusters were identified, each rep-
resenting a different level of environmental quality. Cluster I (less polluted—LP) includes
points 1, 3, and 8, which exhibited favorable physicochemical conditions such as low-to-
moderate levels of electrical conductivity, BOD5, COD, and turbidity. These sites also
Sustainability 2025, 17, 3684 16 of 32

Based on the dendrogram (Figure 6), three distinct clusters were identified, each
representing a different level of environmental quality. Cluster I (less polluted—LP) in-
cludes points 1, 3, and 8, which exhibited favorable physicochemical conditions such as
low-to-moderate levels of electrical conductivity, BOD5 , COD, and turbidity. These sites
also recorded high scores in ecological indices (ABI, BMWP-Col, IHF, QBR-And), reflecting
better habitat structure and biological integrity. Cluster II (moderately polluted—MP)
comprises points 2, 4, 5, 6, and 7, which showed intermediate conditions. While some sites
within this group exhibited relatively high values in individual structural indices—such as
a high IHF value at point 6 and high QBR-And value at point 2—moderate concentrations of
BOD5 , COD, and phosphates were also recorded, indicating mixed influence from natural
and anthropogenic sources. Cluster III (highly polluted—HP) includes points 9, 10, and 11,
which presented high values of electrical conductivity, BOD5 , COD, turbidity, TDS, and
fecal coliforms. These sites also had low scores in both biotic and structural indices.

3.4.3. Spatial Variation in Environmental Quality by Cluster Group

Using the defined groups, boxplots were generated to visualize the results, revealing
significant variations in the environmental quality of the sampling sites. The variations in
each parameter are reflected in Figure 7, highlighting the differences among the groups (LP,
MP, HP).
Apparent color values showed statistically significant differences among clusters
(p < 0.05). The highest concentrations were recorded in the highly polluted (HP) group,
with median values around 215 CU, while both the less polluted (LP) and moderately
polluted (MP) clusters exhibited markedly lower and comparable values, with medians
near 45 and 60 CU, respectively.
For true color, significant differences were also observed (p < 0.05). The HP cluster
presented the lowest median values (≈25 CU), contrasting with the LP cluster (≈55 CU),
which had higher concentrations. The MP cluster displayed intermediate values (≈65 CU)
that did not differ significantly from either the LP or HP clusters.
Electrical conductivity varied significantly across clusters (p < 0.05), with the HP
cluster exhibiting the highest median value (≈430 µS/cm). In contrast, the LP and MP
clusters showed lower and statistically similar values (≈140 µS/cm and ≈120 µS/cm,
respectively).
Biochemical oxygen demand (BOD5 ) did not differ significantly among the clusters
(p > 0.05). Median concentrations remained elevated in all three groups, with values around
29 mg/L (LP), 35 mg/L (MP), and 31 mg/L (HP), consistently exceeding the national
regulatory standard of 20 mg/L.
Similarly, chemical oxygen demand (COD) showed no statistically significant differ-
ences (p > 0.05), with median values of 57 mg/L (LP), 68 mg/L (MP), and 61 mg/L (HP).
These results also surpassed the TULSMA threshold of 40 mg/L.
In the case of fluorides, the HP group exhibited significantly higher concentrations
(p < 0.05), with a median close to 0.95 mg/L. The LP and MP clusters reported lower and
comparable values (≈0.28 mg/L and ≈0.40 mg/L, respectively), all within the permissible
limit of 1.60 mg/L.
Phosphate levels did not differ significantly across clusters (p > 0.05), with median
values of approximately 1.1 mg/L (LP), 1.0 mg/L (MP), and 3.0 mg/L (HP). Although
higher in the HP cluster, the differences were not statistically significant.
Sustainability 2025, 17, 3684
Sustainability 2025, 17, x FOR PEER REVIEW 17 of 17
34 of 32

Figure 7. Cont.
Sustainability 2025, 17, 3684
Sustainability 2025, 17, x FOR PEER REVIEW 18 of1834of 32

Figure7.7. Boxplots
Figure physicochemicaland
Boxplots of physicochemical andmicrobiological
microbiological parameters
parameters by by cluster
cluster classification.
classification.
Results
Resultsofofthe
theTukey
Tukeypost
post hoc
hoc test (p
(p <<0.05)
0.05)are
areshown.
shown.Clusters
Clusters sharing
sharing thethe same
same letter
letter do not
do not differ
differ
significantly.
significantly.Symbols
Symbols “o”
“o” represent mild outliers,
represent mild outliers,and
andsymbols
symbols“*”
“*”represent
represent extreme
extreme outliers.
outliers.

Nitrate
Apparentconcentrations
color values remained below the significant
showed statistically regulatorydifferences
threshold of 5 mg/L
among at all (p
clusters sites,
with no significant differences observed among clusters (p > 0.05). Median values
< 0.05). The highest concentrations were recorded in the highly polluted (HP) group, with were
approximately 0.6 mg/L
median values around 215(LP), 0.4 mg/L
CU, while (MP),
both the less and 2.5 mg/L
polluted (HP).
(LP) and In contrast,
moderately nitrite
polluted
concentrations differed significantly (p < 0.05), with the highest median value recorded in
Sustainability 2025, 17, 3684 19 of 32

the HP group (≈0.009 mg/L). The LP and MP clusters exhibited lower and statistically
similar values around 0.005 mg/L, all within the acceptable limit of 0.050 mg/L.
Dissolved oxygen (% saturation) also showed statistically significant variation
(p < 0.05). The MP group recorded the lowest values (median ≈ 58%), while the LP
cluster had the highest (≈61%). The HP cluster exhibited intermediate values (≈60%), not
differing significantly from either group.
Regarding pH, significant differences were identified (p < 0.05). The LP cluster showed
slightly higher median values (≈7.65) compared to the MP (≈7.55) and HP (≈7.70) clusters,
both of which remained within the acceptable range of 6-to-9.
Salinity did not present statistically significant differences across clusters (p > 0.05).
Values remained relatively low, with medians around 0.07 ppt (LP), 0.18 ppt (MP), and
0.22 ppt (HP).
Total dissolved solids (TDS) differed significantly between clusters (p < 0.05). The HP
group displayed the highest median value (≈220 mg/L), while the LP and MP clusters
recorded lower and comparable concentrations (≈70 mg/L and ≈60 mg/L, respectively),
all below the regulatory limit of 500 mg/L.
Sulfates were significantly higher in the HP cluster (p < 0.05), with median values
around 10 mg/L. In contrast, the LP and MP clusters registered negligible concentrations,
with no statistical difference between them.
Water temperature also differed significantly (p < 0.05). The HP cluster presented the
highest median temperature (≈16 ◦ C), whereas the LP and MP clusters recorded cooler
conditions (≈12.5 ◦ C and ≈12 ◦ C, respectively), which did not differ significantly from
each other.
Turbidity showed statistically significant differences among clusters (p < 0.05). The
HP cluster had the highest levels (median ≈ 19 NTU), exceeding the regulatory standard of
10 NTU. The LP and MP clusters reported lower and statistically similar values (≈4 NTU).
Fecal coliform counts did not differ significantly among clusters (p > 0.05). All groups
remained below the national threshold of 200 CFU/100 mL, with median values around
5–10 CFU/100 mL for the LP and MP clusters, and approximately 30 CFU/100 mL for the
HP cluster.
A similar analysis was conducted using the five indices, comparing their results with
the qualitative scale presented in Table 4. These findings are illustrated in Figure 8.
According to the Fluvial Habitat Index (IHF), the LP cluster presented values ranging
from 67 to 72, with a median of 70. MP sites had a slightly wider range (66-to-74), also
with a median of 70. No significant differences were found between these two clusters. In
contrast, the HP cluster recorded lower values, between 58 and 61, with a median of 60,
significantly differing from LP and MP groups (p < 0.05), and indicating a shift from “good”
to “fair” habitat quality.
The QBR-And showed relatively high values for LP and MP clusters. LP sites ranged
from 66 to 83, with a median of 77, while MP sites ranged from 76 to 90, with a median of
80. The HP cluster, however, presented notably lower values (median: 66; range: 63–68).
Despite this trend, statistical analysis did not detect significant differences among the three
groups (p ≥ 0.05), suggesting similar riparian vegetation quality across clusters, although
with observable ecological degradation in HP sites.
In the ABI, LP sites recorded the highest scores, ranging from 43 to 50 (median: 47.5),
followed by HP (median: 35; range: 29–38) and MP (median: 30; range: 20–38) sites. All
three groups were categorized as having “fair” biological quality. Statistically, no significant
differences were found among the clusters (p ≥ 0.05), despite the lower values observed in
MP and HP sites.
Sustainability 2025, 17, 3684 three groups (p ≥ 0.05), suggesting similar riparian vegetation quality across clusters,
20 alt-
of 32
hough with observable ecological degradation in HP sites.

Figure 8. Boxplot of ecological indices categorized by cluster classification. Results of the Tukey post
Figure 8. Boxplot of ecological indices categorized by cluster classification. Results of the Tukey post
hoc test (p < 0.05) are shown. Clusters sharing the same letter do not differ significantly. Symbol “o”
hoc test (p < 0.05) are shown. Clusters sharing the same letter do not differ significantly. Symbol ʺoʺ
represent mild outliers.
represent mild outliers.
The BMWP-Col exhibited clearer contrasts. LP sites showed values between 61 and
In the ABI,
70 (median: 65),LPplacing
sites recorded
them inthethehighest
“good”scores,
water ranging from 43 toMP
quality category. 50 (median:
sites had 47.5),
lower
followed by HP (median: 35; range: 29–38) and MP (median: 30; range:
scores, ranging from 30 to 50 (median: 38), corresponding to “fair” quality, and 20–38) sites.were
All
three groups were
significantly categorized
different from LP assites
having “fair”
(p < 0.05).biological
HP sitesquality.
rangedStatistically,
from 35 to no50,signifi-
with a
cant differences were found among the clusters (p ≥ 0.05), despite the lower
median of 45. These values were intermediate and did not differ significantly from either values ob-
served
LP or MPin MP andsuggesting
sites, HP sites. partial overlap in macroinvertebrate composition and water
quality classification.exhibited clearer contrasts. LP sites showed values between 61 and
The BMWP-Col
70 (median:
Finally,65),
the placing them inrevealed
WQI-TULSMA the “good” water quality
statistically category.
significant MP sites
differences hadclusters.
among lower
scores, ranging from 30 to 50 (median: 38), corresponding to “fair” quality,
LP sites had values between 50 and 57 (median: 53), and MP site values ranged from and were sig-
nificantly different 61),
49 to 68 (median: fromboth
LP sites (p into
falling < 0.05).
theHP sites
“fair” ranged
water fromcategory.
quality 35 to 50, with
The HPa median
cluster
of 45. These values were intermediate and did not differ significantly
showed higher WQI scores, from 75 to 79, with a median of 77, crossing the threshold from either LPinto
or
the “poor” quality classification. These values were significantly different from those in
Sustainability 2025, 17, 3684 21 of 32

LP and MP sites (p < 0.05), indicating a pronounced degradation in physicochemical and

microbiological parameters in HP sites.

4. Discussion
4.1. Physicochemical Parameters
The physicochemical results from the Columbe River micro-watershed revealed ev-
ident spatial patterns of degradation across the fluvial system. The highest BOD5 and
COD concentrations were recorded at P2 (BOD5 = 49.8 mg/L, COD = 99.6 mg/L) and P10
(BOD5 = 30.5 mg/L, COD = 61.0 mg/L), clearly exceeding the thresholds established by the
Ecuadorian TULSMA regulation for aquatic life preservation (20 mg/L and 40 mg/L,
respectively). These elevated values reflect chronic organic pollution, comparable to
those reported in the Chumbao River (Peru) (BOD5 = 105.38 mg/L) [42], and signifi-
cantly higher than in relatively undisturbed Andean rivers, such as the Volcán River (Costa
Rica) (3.93 mg/L) [19] and Yanaquincha River (Ecuador) (0.8 mg/L) [11]. Interestingly, P6
showed moderate BOD5 levels (23.6 mg/L) but was classified as “good” in terms of habitat
quality (IHF = 70), reinforcing the need for integrated, multimetric assessments.
Dissolved oxygen (DO) saturation ranged from 56.8% to 62.2%, well below the 80%
threshold required by TULSMA. These hypoxic conditions likely result from high organic
matter inputs, biological oxygen demand, and low turbulence in altered segments, trends
consistent with observations in other tropical mountain systems [43,44].
The pH values were stable and neutral across all sampling points, ranging from
7.38 to 7.72. This pattern is consistent with values documented in the Orienco and Ichu
streams [44,45], and reflects the buffering effect of bicarbonate weathering in high-altitude
Andean systems [46]. Despite its limited sensitivity to pollution, pH provides insight into
the geochemical stability and resilience of the system.
Electrical conductivity (EC) varied from 98.4 µS/cm at P2 to 473.8 µS/cm at P10, indi-
cating substantial ion enrichment in more impacted sites. These values surpass those found
in minimally disturbed rivers located in Cajas National Park (Ecuador) (70.5 µS/cm) [2]
and approach levels reported in urban-impacted basins like the Tarqui River (Ecuador)
(131.4 µS/cm) [2]. Elevated EC suggests the presence of untreated wastewater, agrochemi-
cal runoff, and leached solutes from deforested soils.
Turbidity peaked at P9 and P10 (18.7 and 20.3 NTU, respectively), exceeding the
10 NTU threshold set by TULSMA for aquatic life protection. These elevated values suggest
substantial sediment input, likely driven by riparian vegetation loss, surface erosion, and
unregulated runoff. Similar turbidity increases have been reported in deforested and
livestock-impacted systems: for example, the Arroyo Grande River in Argentina exhibited
turbidity levels of 59.7 NTU under similar land-use pressures [47]. In the Colombian Andes,
deforestation for cattle grazing has been shown to cause soil compaction, reduce infiltration,
and increase surface runoff, thereby elevating sediment loads in nearby water bodies [48].
Nitrate and nitrite concentrations generally remained within legal limits, with slightly
elevated values at P10 (NO3 − = 4.65 mg/L; NO2 − = 0.011 mg/L). This pattern is consistent
with denitrification processes in oxygen-depleted sediments and plant uptake in riparian
zones [49,50]. Nonetheless, their presence in impacted areas suggests continuous input
from diffuse sources such as agriculture, livestock, and domestic discharges [51].
Elevated phosphate concentrations at sampling points P9 (3.12 mg/L) and P10
(3.03 mg/L), coinciding with increased turbidity and organic matter levels, suggest signifi-
cant phosphorus loading. This phenomenon is commonly observed in tropical watersheds,
where phosphorus attaches to suspended particles, exacerbating eutrophication and dis-
rupting biogeochemical cycles. For instance, studies have shown that phosphorus concen-
trations in water bodies rarely exceed 0.1 mg/L for inorganic phosphorus and 0.5 mg/L
Sustainability 2025, 17, 3684 22 of 32

for total phosphorus [52]. However, during storm events, total particulate phosphorus
(TPP) concentrations can increase significantly, with measurements averaging 118.4 µg P/L,
which is 1.5-times higher than pre-storm riverine concentrations [53]. These elevated phos-
phorus levels, particularly when associated with suspended sediments, can lead to harmful
algal blooms and oxygen depletion, as excessive phosphorus promotes the overgrowth of
algae, subsequently affecting aquatic life and water quality [54].
Fecal coliform concentrations peaked at point P9, reaching 140 CFU/100 mL, sub-
stantially higher than at the other sampling sites. This elevated value suggests localized
contamination, likely from untreated domestic wastewater or nearby livestock activity.
In comparable rural settings, studies have shown that cattle access to stream banks can
nearly double coliform levels in rivers [55]. In similar rural Andean settings, such as the
Retamales River in Latacunga, reported fecal coliform levels were 66 and 41 CFU/100 mL,
respectively, which are noticeably lower than those recorded at P9 in this study [17].
Hydrological conditions played a relevant role in the spatial distribution of pollutants.
Sites with higher discharge, such as P3 (0.845 m3 /s) and P11 (0.856 m3 /s), did not neces-
sarily present better physicochemical conditions. For instance, P11 exhibited a high EC
and organic load despite its flow rate, illustrating that increased discharge is not sufficient
to offset accumulated pollution [56]. Conversely, low-flow sites such as P1 (0.017 m3 /s)
and P7 (0.065 m3 /s) were more prone to localized pollutant accumulation due to limited
dilution capacity, a common pattern in low-flow systems with restricted turbulence. Dun-
can et al. (2024) examined the effects of droughts and heatwaves on freshwater quality
globally, emphasizing that reduced discharge limits a river’s dilution capacity and can lead
to increased concentrations of pollutants, particularly during extreme climatic events [57].
Similarly, Pakoksung et al. (2025) assessed seasonal water quality dynamics in the Chi-Mun
River Basin, Thailand [58]. They found that contaminant levels were consistently higher
during the dry season, a pattern linked to lower streamflows and reduced dilution [58].

4.2. Aquatic Diversity (ABI and BMWP-Col Indices)

Aquatic macroinvertebrate diversity in the Columbe River micro-watershed, assessed
using the Andean Biotic Index (ABI) and BMWP-Col, revealed a marked gradient of
ecological quality along the longitudinal profile of the watershed. The highest biotic
integrity was observed at sites P3 and P1, with ABI scores of 50 and 48, and BMWP-Col
values of 71 and 65, respectively. These values reflect the presence of pollution-sensitive
taxa such as Perlidae, Baetidae, and Odontoceridae, which are indicators of good water
quality and complex habitat structure [13,59]. Similar macroinvertebrate assemblages have
been reported in reference headwater streams of the Guatapurí River in Colombia (BMWP-
Col = 148) and the Yanaquincha River in the Ecuadorian Amazon (BMWP-Col = 72) [11,12].
In contrast, the most degraded sites—P9, P10, and P11—exhibited reduced biotic
scores, with ABI values below 35 and BMWP-Col values below 50. These stations were
dominated by tolerant taxa such as Chironomidae and Oligochaeta, consistent with high
organic loads and habitat simplification. This assemblage structure mirrors conditions
in heavily impacted systems like the downstream Guatapurí River (Colombia) (BMWP-
Col = 130) [12], Antisana River Basin (ABI = 41) [10], and the lower Paute River, where
sewage inputs and loss of structural complexity contribute to biological degradation [2].
Interestingly, inconsistencies in site classification emerged between the two biotic
indices. For example, P4 and P7 were rated as “good” by the BMWP-Col (scores above
55), but received “poor” ratings under the ABI (scores below 30). This divergence is
attributable to the differing design and sensitivity of the indices: the ABI prioritizes taxa
endemic to high-elevation Andean systems, assigning higher weight to sensitive-but-less-
diverse communities, while the BMWP-Col emphasizes overall family richness. Such
Sustainability 2025, 17, 3684 23 of 32

discrepancies have been documented in other Andean catchments, including the Antisana
and Cunas rivers, where index misalignment was linked to localized taxonomic dominance
by moderately tolerant families [10,18].
Point P10 further exemplifies this pattern, as it was classified as “poor” by the WQI
(score = 79) but recorded a moderate BMWP-Col score (50). This suggests either a lag in
biological response or temporary recovery mechanisms within the macroinvertebrate com-
munity [60]. Additionally, hydrological conditions such as localized turbulence or substrate
heterogeneity may support sensitive taxa even under moderate pollution levels [61].
Conversely, P8, a site with low pollution input and intermediate flow (0.156 m3 /s),
exhibited a lower ABI score (41) than expected for its chemical and structural conditions.
This may be due to natural limitations in taxonomic richness associated with high-elevation
streams, where faunal turnover and reduced colonization opportunities constrain biotic
scores despite relatively intact habitats [1,13].
Overall, the biotic indices provided consistent evidence of ecological degradation in
downstream segments and highlighted the role of anthropogenic stressors such as organic
pollution, erosion, and riparian disturbance in shaping macroinvertebrate assemblages.
However, the influence of natural variability—particularly hydrological dynamics—must
be considered in interpreting index performance. Sites with moderate flow (e.g., P3)
and minimal disturbance supported diverse and sensitive communities, reinforcing the
importance of hydromorphological stability in maintaining ecological integrity. These
findings are consistent with broader observations in the tropical Andes, where stream
velocity, turbulence, and substrate complexity are critical in sustaining benthic diversity. In
particular, Vázquez et al. (2020) [62] found that macroinvertebrate distribution in Andean
rivers is closely linked to hydraulic factors such as flow velocity, turbulence, substrate
complexity, and algal coverage. Their habitat suitability models, developed for key taxa in
the Yanuncay River (Ecuador), revealed that sensitive groups like Baetodes, Simulium, and
Anacroneuria thrive in fast, complex microhabitats. These findings highlight how changes
in flow regimes or habitat simplification can significantly impact ecological integrity, even
in chemically pristine streams.

4.3. Fluvial Habitat Structure (IHF and QBR Indices)

The fluvial habitat conditions in the Columbe River micro-watershed varied consid-
erably across sampling points, with clear signs of structural degradation associated with
human pressures. The Fluvial Habitat Index (IHF) ranged from 58 to 73, with the lowest
scores recorded at P9 and P10. These values reflect habitat simplification due to sedimenta-
tion, riparian clearance, and flow homogenization. Similar IHF scores have been reported
in degraded downstream segments of the Nonguén River in Chile (IHF = 45) [63] and the
Paute River in Ecuador (IHF = 39) [2], where channelization and vegetation loss contribute
to reduced physical complexity.
In contrast, the highest IHF values were observed at points P2, P3, and P6 (≥70), indi-
cating more heterogeneous habitats with varied substrates, riffle frequency, and channel
morphology. These sites featured better-developed instream habitats, including gravel bars,
root systems, and variable depths, characteristics known to favor sensitive macroinverte-
brate taxa [19,64]. The findings mirror upstream reaches of the Guatape (IHF = 79.7) and
Volcán (IHF = 52) rivers, where similarly high IHF values have been linked to the presence
of intact channel structures and well-preserved riparian buffers [19,64].
Riparian quality, assessed through the QBR-And, ranged from “poor” (score = 40) at
P4 and P10 to “good” (score = 90) at P3. Lower values were associated with sites exhibiting
extensive bank modification and vegetation replacement by crops or pasture. These patterns
align with findings in the Guatapurí and Cunas rivers, where degraded riparian zones were
Sustainability 2025, 17, 3684 24 of 32

linked to higher turbidity, elevated temperatures, and loss of habitat continuity [12,18].
Conversely, sites with dense canopy cover and native vegetation showed better biotic scores,
confirming the critical role of riparian corridors in maintaining microclimatic stability and
buffering surface runoff. This is consistent with broader ecological principles and recent
studies demonstrating that riparian buffer width and vegetation structure significantly
influence ecosystem attributes such as shading, organic matter input, sediment control, and
biodiversity [65]. For instance, in Swedish boreal and temperate forests streams bordered by
wider riparian buffers exhibited higher canopy cover and improved proxies for ecological
functions like food provision and temperature regulation, especially in comparison with
harvested sites lacking protective vegetation [66].
Despite some structural quality in specific points (e.g., P6, QBR-And = 78; IHF = 70), in-
consistencies with chemical degradation indicators (e.g., high BOD5 and turbidity) suggest
that habitat condition alone does not guarantee ecological integrity when pollutant loads
are excessive. This decoupling effect has also been observed in the Talala River (Ecuador),
where persistent organic pollution masked the ecological benefits of partial vegetation
recovery [8].
Hydrologically, sites with moderate-to-low discharge (e.g., P6 = 0.419 m3/s; P3 = 0.845 m3/s)
supported more complex instream structures, possibly due to reduced erosive forces and a
greater balance between sediment deposition and habitat formation. In contrast, high-flow
points such as P11 (0.856 m3 /s) showed low IHF scores (60), suggesting that increased
discharge—when coupled with upstream disturbance—can lead to substrate instability
and channel erosion. This pattern is consistent with findings by K˛edzior et al. (2022) [67],
who demonstrated that river incision significantly reduces the area of optimal habitat
for macroinvertebrates. In such cases, even high environmental flow values may be
insufficient to sustain biological integrity, as intense morphological degradation disrupts
habitat complexity and limits faunal colonization.

4.4. Correlation Patterns Between Environmental and Biological Metrics

The correlation analysis (Figure 5) revealed statistically significant relationships
(p < 0.01) between various physicochemical, structural, and biotic variables, offering
insights into shared pollution sources and ecological responses in the Columbe River
micro-watershed.
Apparent color showed strong positive correlations with electrical conductivity (EC;
r = 0.97), fluorides (r = 0.91), total dissolved solids (TDS; r = 0.97), sulfates (r = 0.95), and
turbidity (r = 0.97). These results indicate that apparent color in the Columbe River micro-
watershed is closely linked to the presence of dissolved and suspended pollutants, many of
which likely originate from domestic wastewater, erosion processes, and agricultural runoff.
Similar associations have been observed in rivers of the Ecuadorian Amazon, where color
metrics increased in parallel with ionic content and particulate load. Likewise, in the Talala
River (Ecuador), Spearman correlation analysis from the period 2018–2019 revealed that
apparent color was moderately associated with turbidity (r = 0.6), electrical conductivity
(r = 0.4), iron (r = 0.4), and total coliforms (r = 0.6) [68], which also reflect pollutant inputs
from erosion and untreated wastewater discharges.
The strong correlations between electrical conductivity (EC) and TDS (r = 0.99), sulfates
(r = 0.93), and turbidity (r = 0.96) observed in the Columbe River align with findings from
other studies. Thirumalini and Joseph (2008) [69] reported that EC explained up to 96%
of TDS variability in natural waters, underscoring its usefulness as a proxy for ionic
concentration. Similarly, Scott and Haggard (2019) [70] found elevated levels of TDS,
turbidity, and sulfates in the downstream segments of the West Fork White River (USA),
linked to anthropogenic land use.
Sustainability 2025, 17, 3684 25 of 32

The strong correlations between fluoride and sulfate (r = 0.95), and between fluoride
and turbidity (r = 0.92), observed in the Columbe River suggest co-occurrence linked to
shared sources such as leachates, erosion, and altered volcanic substrates. In the Cauca
region (Colombia), fluoride concentrations reached up to 0.83 mg/L in the Cauca River,
attributed to volcanic geology and surface runoff [71]. Similarly, in the Elqui River (Chile),
sulfate-rich waters in the Andean zone reflected the oxidation of sulfide-bearing rocks and
hydrothermal alteration, intensified by mining and weathering [72].
TDS showed strong positive correlations with sulfates (r = 0.99), temperature (r = 0.93),
and turbidity (r = 0.96) in the Columbe River, highlighting the combined influence of dis-
solved and suspended matter in degraded reaches. While similar associations between TDS
and electrical conductivity were observed in the Canuto (r = 0.95) and Carrizal (r = 0.99)
rivers in coastal Ecuador, the relationship between TDS and temperature differed across
sites [73]. In Canuto, temperature showed a weak positive correlation with TDS (r = 0.30),
whereas in Carrizal the correlation was moderate and negative (r = −0.36). These con-
trasting patterns may reflect site-specific hydrological or thermal conditions influencing
solute concentration dynamics, such as differential groundwater inputs, shading, or flow
regulation.
Biochemical oxygen demand (BOD5 ) and chemical oxygen demand (COD) showed a
very strong correlation (r = 0.99) in the Columbe River, confirming their joint reliability as
indicators of organic pollution. BOD5 reflects microbial respiration driven by biodegradable
organic matter, while COD includes both biodegradable and non-biodegradable oxidizable
substances. This high association aligns with findings from wastewater treatment plants in
Egypt, where the highest reported BOD5 /COD biodegradability index reached 0.88 in El
Beheira governorate, illustrating a close relationship between both parameters under high
organic load conditions [74].
The Fluvial Habitat Index (IHF) showed statistically significant negative correlations
of moderate-to-strong intensity with several physicochemical variables: apparent color
(r = −0.88), EC (r = −0.87), fluorides (r = −0.86), phosphates (r = −0.82), TDS (r = −0.87),
sulfates (r = −0.86), temperature (r = −0.81), and turbidity (r = −0.82). These results
underscore the inverse relationship between structural habitat quality and physicochemical
degradation. Similar patterns have been reported by Echeverría-Sáenz et al. (2022) [19] and
Munné et al. (2003) [38], who highlighted that increased sedimentation, thermal load, and
ionic concentration contribute to habitat homogenization and degradation.
Importantly, biotic indices demonstrated internal consistency. The Andean Biotic Index
(ABI) showed a strong positive correlation with the BMWP-Col (r = 0.91), confirming a high
degree of congruence in their classification of ecological status based on macroinvertebrate
assemblages. This supports earlier findings by Ríos-Touma et al. (2014) [13], who noted
that while each index has its strengths and taxonomic nuances, both tend to converge in
detecting pollution gradients when family richness is adequate.
The Water Quality Index (WQI) showed a moderate negative correlation with IHF
(r = −0.77), supporting the notion that declines in structural complexity coincide with
worsening chemical quality. This pattern is consistent with findings from the Maranhão
Amazon, where Braga et al. (2022) [75] reported that sites with elevated turbidity, nutrient
loads, and microbiological contamination—particularly during the rainy season—were
associated with reduced WQI scores, as revealed through multivariate statistical analysis.
Although habitat variables were not directly measured, the clustering of degraded chemical
conditions in specific sites suggests underlying structural deterioration. Similarly, in the
Paute River Basin (Ecuador), Sotomayor et al. (2016) [76] found that riparian vegetation and
streambed heterogeneity were the strongest predictors of better macroinvertebrate-based
Sustainability 2025, 17, 3684 26 of 32

water quality classes, while poor habitat structure coincided with higher levels of pollutants
such as fecal coliforms and thermal stress.

4.5. Spatial Distribution of Physicochemical and Biological Indices

The hierarchical cluster analysis clearly distinguished three ecological conditions
across the Columbe River micro-watershed: less polluted (LP: P1, P3, P8), moderately
polluted (MP: P2, P4, P5, P6, P7), and highly polluted (HP: P9, P10, P11). This classification
was statistically supported and allowed the spatial differentiation of the system based on
pollution intensity, habitat integrity, and biological responses.
LP sites exhibited the best ecological conditions. They were characterized by high ABI
scores (median = 47.5), elevated BMWP-Col values (median = 65), low turbidity (<5 NTU),
and good fluvial habitat structure (IHF median = 70). For example, P3, with a flow of
0.845 m3 /s, registered some of the highest ABI (50) and IHF (73) values, suggesting that a
combination of moderate hydrological stability, minimal pressure, and habitat complexity
can support sensitive macroinvertebrate assemblages even under relatively low base flows.
These conditions resemble those described in upstream segments of the Guatapurí River
(BMWP-Col = 148, QBR = 80) [12] and the Yanaquincha River (BMWP-Col = 72) [11],
confirming their status as local reference conditions.
MP sites showed intermediate values and mixed signals of degradation. For example,
P6 had an IHF of 70 and QBR-And of 78, suggesting relatively good habitat structure, but
simultaneously presented BOD5 = 23.6 mg/L and moderate turbidity. Similarly, P2 had the
highest COD (99.6 mg/L) and BOD5 (49.8 mg/L), despite moderate IHF (68), underscoring
the complex dynamics between structural features and pollution inputs. These findings
reflect partial degradation, often due to diffuse pollution and localized habitat disturbance.
Comparable transitional conditions have been reported in the mid-reaches of the Guatape
and Cunas rivers, where moderate flows and agricultural runoff produce heterogeneity in
ecological indicators [18,64].
HP sites were consistently associated with high pollutant concentrations and biological
impoverishment. For example, P10 and P11 exhibited elevated BOD5 (30.5 and 26.1 mg/L),
COD (61.0 and 54.0 mg/L), turbidity (>18 NTU), and conductivity (>400 µS/cm), along with
low IHF scores (58 and 60). Biologically, these sites were dominated by tolerant taxa such as
Chironomidae and Oligochaeta, and had ABI scores below 35, indicating a severely altered
ecological status. Similar conditions were reported in the Paute River Basin (Ecuador),
where the most degraded sites (C3 class) were associated with low habitat quality and
a simplified benthic community dominated by pollution-tolerant groups, underpinned
largely by the loss of riparian vegetation and instream heterogeneity [76]. In the Portoviejo
River (Ecuador), Nguyen et al. (2015) [77] found that macroinvertebrate community com-
position shifted markedly in response to rising conductivity, with tolerant taxa becoming
dominant at sites exceeding 1430 µS/cm, supporting the link between ionic concentration
and biological simplification. Likewise, Banda et al. (2022) [78] observed that high con-
ductivity and organic enrichment in a tropical wetland led to reductions in community
diversity and the prevalence of tolerant macroinvertebrate assemblages. These consistent
findings across different systems reinforce the interpretation that elevated BOD5 , COD,
turbidity, and conductivity—coupled with low structural integrity—underpin ecological
degradation and favor dominance by tolerant taxa, as observed in the Columbe River.
Interestingly, P11—despite its relatively high discharge (0.856 m3 /s)—remained in
the high-pollution group, underscoring that increased flow alone does not mitigate the
impacts of pollution when structural degradation and external inputs are severe. This
finding aligns with the work of Simon and Rinaldi (2006) [79], who demonstrated that
in incised or hydromorphologically unstable channels, increases in stream power due to
Sustainability 2025, 17, 3684 27 of 32

higher flows can actually exacerbate erosion and habitat loss when boundary resistance is
low. Similarly, Santikari and Murdoch (2020) [80] observed that in urbanizing watersheds,
elevated stormflows—often several times greater than in undisturbed systems—did not
prevent sediment surges and water quality deterioration, especially when vegetation and
soil stability were compromised. Moreover, Cashman et al. (2023) [81] emphasized that
degraded physical habitat is not merely a sediment issue, but is tightly linked to altered
flow regimes. Their findings indicate that even in systems with high discharge, the absence
of hydromorphic heterogeneity and riparian stability leads to declines in habitat quality.
Taken together, these studies support the notion that flow magnitude, in isolation, is
insufficient to buffer the ecological impacts of structural and chemical degradation, an
interpretation confirmed by the impaired status of P11 in the Columbe River.
In contrast, P1, with the lowest flow (0.017 m3 /s), maintained high biological and
structural index values, although it showed elevated BOD5 (27.8 mg/L). This suggests that
headwater segments, despite their lower dilution capacity, can maintain ecological integrity
when pressure levels are minimal and riparian buffers are preserved. Similar dynamics
were observed in the upstream Volcán and Guatapurí rivers, where intact vegetation and
minimal land use resulted in higher ecological scores despite natural limitations in flow
and substrate availability [12,19].

4.6. Implications for Sustainability and Integrated Watershed Management

The findings from the Columbe River micro-watershed highlight the urgent need for
integrated watershed management in high-Andean regions undergoing rapid land-use
transformation. The observed decline in water quality, habitat structure, and macroinver-
tebrate diversity reflects broader sustainability challenges common in mountain systems
where environmental regulation is often limited or poorly enforced. Similar conclusions
have been drawn in other highland regions. For example, Haq et al. (2021) [82] demon-
strated that riparian zones in the Kashmir Himalaya sustain high floristic and functional
diversity, which is essential for ecological resilience. Likewise, Mosquera et al. (2023) [83]
emphasized that while páramo ecosystems are critical for hydrological regulation, they
lack integrated research on vegetation–water interactions, highlighting the need for inter-
disciplinary management strategies.
The relationships observed in the Columbe River micro-watershed between land use
and aquatic ecosystem health are consistent with broader empirical evidence. Cortes et al.
(2013) [84] showed that land-use patterns—particularly urbanization at local scales—are
strong predictors of ecological quality in rivers, as reflected by macroinvertebrate-based
bioindicators. Their study highlighted the importance of integrating biotic responses with
spatially explicit land-use data to guide effective river management. Similarly, Brumberg
et al. (2021) [85] found that riparian buffer length, more than width, had the greatest
influence on water quality in agricultural landscapes of southern Costa Rica, suggesting
that even narrow, but continuous, riparian buffers can significantly improve water quality
where full-width restoration is impractical.
Beyond biophysical strategies, institutional mechanisms such as payment for ecosys-
tem services (PES) offer promising tools to incentivize upstream conservation. Izquierdo-
Tort et al. (2022) [20], in a randomized trial in Mexico, showed that requiring full-parcel en-
rollment in PES contracts reduced deforestation by 41% and quadrupled cost-effectiveness
compared to traditional schemes. These findings underscore how strategic adjustments in
PES design can substantially enhance environmental outcomes. Taken together, these stud-
ies reinforce the need to combine ecological indicators, spatial planning, and institutional
innovation to advance sustainability in rapidly transforming Andean watersheds.
Sustainability 2025, 17, 3684 28 of 32

Although this study offers a detailed assessment of ecological integrity and water
quality in a high-Andean micro-watershed, certain limitations should be acknowledged.
First, macroinvertebrate sampling was conducted during a single season, which may not
fully capture temporal variability in community composition influenced by hydrological
fluctuations (e.g., [1,2]). Additionally, the use of indices such as the ABI and BMWP-Col—
although regionally adapted—relies on fixed tolerance scores that may oversimplify the
ecological responses to multiple and interacting stressors (e.g., [3,4]). Moreover, while the
selection of 11 sampling points aimed to ensure representativeness, the spatial coverage
may not fully reflect the heterogeneity of the watershed, particularly in transitional zones
or ephemeral tributaries.
Future research should incorporate multi-seasonal monitoring to better account for
seasonal ecological variation and improve the reliability of biotic assessments (e.g., [5]).
Integrating molecular tools, such as DNA barcoding, could enhance taxonomic resolution
and enable the detection of cryptic diversity often overlooked by conventional methods [6].
Modeling scenarios involving land-use change, climate variability, and restoration actions
may also provide strategic insights for long-term watershed planning [7]. Furthermore,
expanding participatory monitoring with local communities could enhance data coverage
and foster stronger stakeholder involvement in conservation strategies [8].

5. Conclusions
This study provides clear evidence that anthropogenic pressures—particularly agri-
culture, livestock, and untreated domestic discharges—are major drivers of ecological
degradation in the Columbe River micro-watershed. These pressures contribute to the
physicochemical alteration in aquatic systems and are closely linked to reduced macroin-
vertebrate diversity, declining water quality, and deteriorated fluvial habitat structure.
The results show that elevated concentrations of BOD5 , COD, and electrical conduc-
tivity, alongside low values of habitat quality indices (IHF and QBR-And), are strongly
associated with the dominance of tolerant taxa such as Oligochaeta and Chironomidae.
This shift in community composition reflects not only site-specific disturbances but also
cumulative basin-wide impacts that compromise ecosystem functionality.
The combined degradation of chemical and structural conditions leads to ecological
fragmentation and loss of biodiversity in high-Andean aquatic systems. These findings
underscore the need for fluvial restoration strategies that are grounded in empirical data
and consider both ecological and social dimensions. Restoring habitat complexity, improv-
ing water quality, and enhancing ecological connectivity will be essential to safeguard
the ecological integrity and service provision of these vulnerable mountain ecosystems.
In this context, the adoption of integrated watershed management approaches supports
long-term sustainability by aligning conservation actions with land-use planning, local
livelihoods, and climate resilience, which are fundamental components for advancing
Sustainable Development Goals (SDGs) in rural Andean regions.

Author Contributions: A.A.B.-D.: Writing—review and editing, Methodology, Investigation, Re-

sources, Data curation. C.S.: Writing—original draft, Conceptualization, Methodology, Formal
analysis, Investigation, Visualization, Supervision, Project administration. F.A.-A.: Writing—review
and editing, Formal analysis, Investigation, Data curation. A.I.K.: Writing—review and editing,
Software, Validation, Formal analysis, Visualization. E.V.S.: Writing—review and editing, Validation,
Resources, Supervision, Funding acquisition. All authors have read and agreed to the published
version of the manuscript.

Funding: This research received no external funding.

Sustainability 2025, 17, 3684 29 of 32

Data Availability Statement: The datasets used and/or analyzed during the current study are
available from the corresponding author upon reasonable request.

Conflicts of Interest: The authors declare no conflicts of interest.

References
1. Villamarín, C.; Prat, N.; Rieradevall, M. Physical, chemical and hydromorphological characterization of Ecuador and Peru tropical
highland Andean rivers. Lat. Am. J. Aquat. Res. 2014, 42, 1072–1086. [CrossRef]
2. Hampel, H.; Vázquez, R.F.; González, H.; Acosta, R. Evaluating the ecological status of fluvial networks of tropical Andean
catchments of Ecuador. Water 2023, 15, 1742. [CrossRef]
3. Fierro, P.; Bertrán, C.; Tapia, J.; Hauenstein, E.; Peña-Cortés, F.; Vergara, C.; Cerna, C.; Vargas-Chacoff, L. Effects of local land-use
on riparian vegetation, water quality, and the functional organization of macroinvertebrate assemblages. Sci. Total Environ. 2017,
609, 724–734. [CrossRef] [PubMed]
4. Rodríguez-Morales, M.; Acevedo-Novoa, D.; Machado, D.; Ablan, M.; Dugarte, W.; Dávila, F. Ecohydrology of the Venezuelan
páramo: Water balance of a high Andean watershed. Plant Ecol. Divers. 2019, 12, 573–591. [CrossRef]
5. Espinosa, J.; Rivera, D. Variations in water resources availability at the Ecuadorian páramo due to land-use changes. Environ.
Earth Sci. 2016, 75, 1173. [CrossRef]
6. Patiño, S.; Hernández, Y.; Plata, C.; Domínguez, I.; Daza, M.; Oviedo-Ocaña, R.; Ochoa-Tocachi, B.F. Influence of land use on
hydro-physical soil properties of Andean páramos and its effect on streamflow buffering. Catena 2021, 202, 105227. [CrossRef]
7. Villa-Achupallas, M.; Rosado, D.; Aguilar, S.; Galindo-Riaño, M.D. Water quality in the tropical Andes hotspot: The Yacuambi
river (southeastern Ecuador). Sci. Total Environ. 2018, 633, 50–58. [CrossRef]
8. Salazar Flores, C.; Zambrano, C.; Parreno, C. Microbiological assessment of the quality of water supplied to the Latacunga water
treatment plant (Republic of Ecuador). Int. Res. J. 2023, 2, 128. [CrossRef]
9. Campo, Y.M.; Rebolledo, M.I.C.; Londoño, A.M.J. Comparison of water quality between two Andean rivers by using the
BMWP/COL and ABI indices. Acta Biol. Colomb. 2019, 24, 1–9. [CrossRef]
10. Cabrera-García, S.; Goethals, P.L.M.; Lock, K.; Domínguez-Granda, L.; Villacís, M.; Galárraga-Sánchez, R.; Van der Heyden, C.;
Forio, M.A.E. Taxonomic and feeding trait-based analysis of macroinvertebrates in the Antisana River Basin (Ecuadorian Andean
Region). Biology 2023, 12, 1386. [CrossRef]
11. Vargas-Tierras, T.; Suárez-Cedillo, S.; Morales-León, V.; Vargas-Tierras, Y.; Tinoco-Jaramillo, L.; Viera-Arroyo, W.; Vásquez-Castillo,
W. Ecological River Water Quality Based on Macroinvertebrates Present in the Ecuadorian Amazon. Sustainability 2023, 15, 5790.
[CrossRef]
12. Herrera-Martínez, J.R.; Navarro-Sining, B.A.; Torres-Cervera, K.P.; Martínez-García, N.; Royero-Ibarra, A.; Cahuana-Mojica, A.
Determinación de los índices BMWP/COL, QBR, IHF e ICO en Valledupar, Colombia. Rev. Politéc. 2022, 18, 98–109. [CrossRef]
13. Ríos-Touma, B.; Acosta, R.; Prat, N. The Andean Biotic Index (ABI): Revised tolerance to pollution values for macroinvertebrate
families and index performance evaluation. Rev. Biol. Trop. 2014, 62, 249–273. [CrossRef]
14. Roldán, G. La Bioindicación de la Calidad del Agua en Colombia: Propuesta para el uso del Método BMWP-COL; Editorial Universidad
de Antioquia: Medellín, Colombia, 2003.
15. Pardo, I.; Álvarez, M.; Casas, J.J.; Moreno, J.L.; Vivas, S.; Bonada, N.; Alba-Tercedo, J.; Jaimez-Cuéllar, P.; Moya, G.; Prat, N.; et al.
El hábitat de los ríos mediterráneos: Diseño de un índice de diversidad de hábitat. Limnetica 2002, 21, 115–132. [CrossRef]
16. Acosta, R.; Ríos, B.; Rieradevall, M.; Prat, N. Propuesta de un protocolo de evaluación de la calidad ecológica de ríos andinos
(C.E.R.A.) y su aplicación en dos cuencas en Ecuador y Perú. Limnetica 2009, 28, 35–64. [CrossRef]
17. Salazar Flores, C.A.; Kurbatova, A.I.; Mikhaylichenko, K.Y.; Barannikova, S.I. Comprehensive water quality assessment of surface
sources in the city of Latacunga and the canton Pedro Vicente Maldonado in Ecuador. RUDN J. Ecol. Life Saf. 2023, 31, 251–264.
[CrossRef]
18. Custodio, M.; Chávez, E. Quality of the aquatic environment of high Andean rivers evaluated through environmental indicators:
A case of the Cunas River, Peru. Ingeniare 2019, 27, 396–409. [CrossRef]
19. Echeverría-Sáenz, S.; Ugalde-Salazar, R.; Guevara-Mora, M.; Quesada-Alvarado, F.; Ruepert, C. Ecological Integrity Impairment
and Habitat Fragmentation for Neotropical Macroinvertebrate Communities in an Agricultural Stream. Toxics 2022, 10, 346.
[CrossRef]
20. Izquierdo-Tort, S.; Jayachandran, S.; Saavedra, S. Redesigning payments for ecosystem services to increase cost-effectiveness. Nat.
Commun. 2024, 15, 9252. [CrossRef]
21. Vasseur, L.; Andrade, A. Using the Red List of Ecosystems and the Nature-based Solutions Global Standard as an integrated
process for climate change adaptation in the Andean high mountains. Philos. Trans. R. Soc. B 2024, 379, 20220326. [CrossRef]
22. GAD de la Parroquia Columbe. Plan de Desarrollo y Ordenamiento Territorial de la Parroquia Columbe 2019–2023. Available
online: https://es.slideshare.net/slideshow/pdotcolumbe2020pdf/252797787 (accessed on 25 January 2025).
Sustainability 2025, 17, 3684 30 of 32

23. GAD de la Provincia de Chimborazo. Plan de Desarrollo y Ordenamiento Territorial de la Provincia de Chimborazo 2019–2023.
Available online: https://chimborazo.gob.ec/principal/wp-content/uploads/2022/06/PDOT.pdf (accessed on 24 January 2025).
24. Santillán, G. Macroinvertebrados Acuáticos Como Bioindicadores de Calidad de Agua en la Microcuenca Columbe, Cantón Colta.
Master’s Thesis, Universidad Nacional de Chimborazo, Riobamba, Ecuador, 2024. Available online: http://dspace.unach.edu.ec/
handle/51000/13571 (accessed on 25 January 2025).
25. Damanik-Ambarita, M.N.; Lock, K.; Boets, P.; Everaert, G.; Nguyen, T.H.T.; Forio, M.A.E.; Musonge, P.L.S.; Suhareva, N.; Bennet-
sen, E.; Landuyt, D.; et al. Ecological water quality analysis of the Guayas River basin (Ecuador) based on macroinvertebrates
indices. Limnologica 2016, 57, 27–59. [CrossRef]
26. Paller, M.H.; Kosnicki, E.; Prusha, B.A.; Fletcher, D.E.; Sefick, S.A.; Feminella, J.W. Development of an Index of Biotic Integrity for
the Sand Hills Ecoregion of the Southeastern United States. Trans. Am. Fish. Soc. 2017, 146, 112–127. [CrossRef]
27. Edegbene, A.O.; Akamagwuna, F.C.; Odume, O.N.; Arimoro, F.O.; Edegbene, O.T.T.; Akumabor, E.C.; Ogidiaka, E.; Kaine, E.A.;
Nwaka, K.H. A Macroinvertebrate-Based Multimetric Index for Assessing Ecological Condition of Forested Stream Sites Draining
Nigerian Urbanizing Landscapes. Sustainability 2022, 14, 11289. [CrossRef]
28. Raven, P.J.; Holmes, N.T.H.; Charrier, P.; Dawson, F.H.; Naura, M.; Boon, P.J. Towards a harmonized approach for hydromorpho-
logical assessment of rivers in Europe: A qualitative comparison of three survey methods. Aquat. Conserv. 2002, 12, 405–424.
[CrossRef]
29. Fernández, D.; Barquín, J.; Raven, P. A review of river habitat characterisation methods: Indices vs. characterisation protocols.
Limnetica 2011, 30, 217–234. [CrossRef]
30. Uddin, M.G.; Nash, S.; Olbert, A.I. A review of water quality index models and their use for assessing surface water quality. Ecol.
Indic. 2021, 122, 107218. [CrossRef]
31. Lukhabi, D.K.; Mensah, P.K.; Asare, N.K.; Pulumuka-Kamanga, T.; Ouma, K.O. Adapted Water Quality Indices: Limitations and
Potential for Water Quality Monitoring in Africa. Water 2023, 15, 1736. [CrossRef]
32. Hanna Instruments. HI98194 Multiparameter Waterproof Meter. Available online: https://www.hannainstruments.co.uk/multi-
parameter-devices/2147-hi-98194-multiparameter-waterproof-meter (accessed on 10 January 2025).
33. Valladares Pogo, V. Evaluación Ecológica del río Columbe. Bachelor’s Thesis, Escuela Superior Politécnica de Chimborazo
(ESPOCH), Riobamba, Ecuador, 2024. Available online: http://dspace.espoch.edu.ec/handle/123456789/23192 (accessed on 25
January 2025).
34. Instituto Privado de Investigación Sobre Cambio Climático (ICC). Manual de Medición de Caudales; Guatemala. 2017. Available
online: https://icc.org.gt/wp-content/uploads/2023/03/064.pdf (accessed on 30 January 2025).
35. Ministerio de Agricultura del Perú. Manual de Hidrometría; Ministerio de Agricultura del Perú: Lima, Perú, 2005.
36. Pence, R.A.; Cianciolo, T.R.; Drover, D.R.; McLaughlin, D.L.; Soucek, D.J.; Timpano, A.J.; Zipper, C.E.; Schoenholtz, S.H.
Comparison of benthic macroinvertebrate assessment methods along a salinity gradient in headwater streams. Environ. Monit.
Assess. 2021, 193, 765. [CrossRef]
37. Getachew, M.; Mulat, W.L.; Mereta, S.T.; Gebrie, G.S.; Kelly-Quinn, M. Refining benthic macroinvertebrate kick sampling protocol
for wadeable rivers and streams in Ethiopia. Environ. Monit. Assess. 2022, 194, 196. [CrossRef]
38. Munné, A.; Prat, N.; Solà, C.; Bonada, N.; Rieradevall, M. A simple field method for assessing the ecological quality of riparian
habitat in rivers and streams: QBR index. Aquat. Conserv. 2003, 13, 147–163. [CrossRef]
39. Instituto Geográfico Militar del Ecuador (IGM). Capas de Información Geográfica del IGM (Codificación UTF-8). Available
online: http://www.geoportaligm.gob.ec/portal/index.php/descargas/cartografia-de-libre-acceso/registro/ (accessed on 27
January 2025).
40. Gaagai, A.; Aouissi, H.A.; Bencedira, S.; Hinge, G.; Athamena, A.; Heddam, S.; Gad, M.; Elsherbiny, O.; Elsayed, S.; Eid, M.H.; et al.
Application of Water Quality Indices, Machine Learning Approaches, and GIS to Identify Groundwater Quality for Irrigation
Purposes: A Case Study of Sahara Aquifer, Doucen Plain, Algeria. Water 2023, 15, 289. [CrossRef]
41. Yang, J.; Lv, J.; Liu, Q.; Nan, F.; Li, B.; Xie, S.; Feng, J. Seasonal and spatial patterns of eukaryotic phytoplankton communities in
an urban river based on marker gene. Sci. Rep. 2021, 11, 23147. [CrossRef] [PubMed]
42. Ramos-Pacheco, B.S.; Choque-Quispe, D.; Ligarda-Samanez, C.A.; Solano-Reynoso, A.M.; Choque-Quispe, Y.; Aguirre Landa,
J.P.; Agreda Cerna, H.W.; Palomino-Rincón, H.; Taipe-Pardo, F.; Zamalloa-Puma, M.M.; et al. Water Pollution Indexes Proposal
for a High Andean River Using Multivariate Statistics: Case of Chumbao River, Andahuaylas, Apurímac. Water 2023, 15, 2662.
[CrossRef]
43. Huang, Y.; Yang, C.; Wen, C.; Wen, G. S-type Dissolved Oxygen Distribution along Water Depth in a Canyon-shaped and Algae
Blooming Water Source Reservoir: Reasons and Control. Int. J. Environ. Res. Public Health 2019, 16, 987. [CrossRef]
44. Sinche, F.; Cabrera, M.; Vaca, L.; Segura, E.; Carrera, P. Determination of the Ecological Water Quality in the Orienco Stream Using
Benthic Macroinvertebrates in the Northern Ecuadorian Amazon. Integr. Environ. Assess. Manag. 2022, 11, e4666. [CrossRef]
Sustainability 2025, 17, 3684 31 of 32

45. Sánchez-Araujo, V.; Portuguez-Maurtua, M.; Palomino-Pastrana, P.; Escobar-Soldevilla, M.; Saez Huaman, W.; Chávez-Araujo, E.;
Contreras López, E. Water Quality Index and Health Risks in a Peruvian High Andean River. Ecol. Eng. Environ. Technol. 2024, 25,
301–315. [CrossRef]
46. Torres, M.A.; West, A.J.; Clark, K.E.; Paris, G.; Bouchez, J.; Ponton, C.; Feakins, S.J.; Galy, V.; Adkins, J.F. The acid and alkalinity
budgets of weathering in the Andes–Amazon system: Insights into the erosional control of global biogeochemical cycles. Earth
Planet. Sci. Lett. 2016, 450, 381–391. [CrossRef]
47. Granitto, M.; Lopez, M.E.; Bursztyn Fuentes, A.L.; Maluendez Testoni, M.C.; Rodríguez, P. Relationship between riparian zones
and water quality in the main watersheds of Ushuaia City, Tierra del Fuego (Argentina). Ecol. Process. 2025, 14, 18. [CrossRef]
48. Vásquez-Velásquez, G. Headwaters Deforestation for Cattle Pastures in the Andes of Colombia and Its Implications for Soils
Properties and Hydrological Dynamic. Open J. For. 2016, 6, 337–347. [CrossRef]
49. Ye, F.; Duan, L.; Sun, Y.; Yang, F.; Liu, R.; Gao, F.; Xu, Y. Nitrogen removal in freshwater sediments of riparian zone: N-loss
pathways and environmental controls. Front. Microbiol. 2023, 14, 1239055. [CrossRef]
50. Weston, N.B.; Troy, C.; Kearns, P.J.; Bowen, J.L.; Porubsky, W.; Hyacinthe, C.; Meile, C.; Van Cappellen, P.; Joye, S.B. Physicochemi-
cal perturbation increases nitrous oxide production from denitrification in soils and sediments. Biogeosciences 2024, 21, 4837–4851.
[CrossRef]
51. Castillo, M.M.; Allan, J.D.; Brunzell, S. Nutrient Concentrations and Discharges in a Midwestern Agricultural Catchment. J.
Environ. Qual. 2000, 29, 1142–1151. [CrossRef]
52. Boyd, C.E. Phosphorus. In Water Quality; Springer: Cham, Switzerland, 2015. [CrossRef]
53. Zhang, M.; Krom, M.D.; Lin, J.; Cheng, P.; Chen, N. Effects of a storm on the transformation and export of phosphorus
through a subtropical river-turbid estuary continuum revealed by continuous observation. J. Geophys. Res. Biogeosci. 2022, 127,
e2022JG006786. [CrossRef]
54. Jin, Z.; Liao, P.; Jaisi, D.P.; Wang, D.; Wang, J.; Wang, H.; Jiang, S.; Yang, J.; Qiu, S.; Chen, J. Suspended phosphorus sustains algal
blooms in a dissolved phosphorus-depleted lake. Water Res. 2023, 241, 120134. [CrossRef]
55. Wilson, J.L.; Everard, M. Real-time consequences of riparian cattle trampling for mobilization of sediment, nutrients and bacteria
in a British lowland river. Int. J. River Basin Manag. 2017, 16, 231–244. [CrossRef]
56. Kowalkowski, T.; Pastuszak, M.; Igras, J.; Buszewski, B. Differences in emission of nitrogen and phosphorus into the Vistula and
Oder basins in 1995–2008—Natural and anthropogenic causes (MONERIS model). J. Mar. Syst. 2012, 89, 48–60. [CrossRef]
57. Graham, D.J.; Bierkens, M.F.P.; van Vliet, M.T.H. Impacts of droughts and heatwaves on river water quality worldwide. J. Hydrol.
2024, 629, 130590. [CrossRef]
58. Pakoksung, K.; Inseeyong, N.; Chawaloesphonsiya, N.; Punyapalakul, P.; Chaiwiwatworakul, P.; Xu, M.; Chuenchum, P. Seasonal
dynamics of water quality in response to land use changes in the Chi and Mun River Basins, Thailand. Sci. Rep. 2025, 15, 7101.
[CrossRef]
59. Jerves-Cobo, R.; Córdova-Vela, G.; Iñiguez-Vela, X.; Díaz-Granda, C.; Van Echelpoel, W.; Cisneros, F.; Nopens, I.; Goethals, P.L.M.
Model-Based Analysis of the Potential of Macroinvertebrates as Indicators for Microbial Pathogens in Rivers. Water 2018, 10, 375.
[CrossRef]
60. Álvarez-Cabria, M.; Barquín, J.; Juanes, J.A. Spatial and seasonal variability of macroinvertebrate metrics: Do macroinvertebrate
communities track river health? Ecol. Indic. 2010, 10, 2. [CrossRef]
61. Heatherly, T., II; Whiles, M.R.; Royer, T.V.; David, M.B. Relationships between water quality, habitat quality, and macroinvertebrate
assemblages in Illinois streams. J. Environ. Qual. 2007, 36, 1653–1660. [CrossRef] [PubMed]
62. Vázquez, R.F.; Vimos-Lojano, D.; Hampel, H. Habitat Suitability Curves for Freshwater Macroinvertebrates of Tropical Andean
Rivers. Water 2020, 12, 2703. [CrossRef]
63. Palma, A.; Figueroa, R.; Ruiz, V.H. Evaluación de ribera y hábitat fluvial a través de los índices QBR e IHF. Gayana (Concepc.) 2009,
73, 57–63. [CrossRef]
64. Galeano-Rendón, E.; Monsalve-Cortés, L.M.; Mancera-Rodríguez, N.J. Evaluación de la calidad ecológica de quebradas andinas
en la cuenca del río Magdalena, Colombia. Rev. UDCA Act. Divulg. Cient. 2017, 20, 413–424.
65. Lind, L.; Hasselquist, E.M.; Laudon, H. Towards ecologically functional riparian zones: A meta-analysis to develop guidelines for
protecting ecosystem functions and biodiversity in agricultural landscapes. J. Environ. Manag. 2019, 249, 109391. [CrossRef]
66. Chellaiah, D.; Kuglerová, L. Are riparian buffers surrounding forestry-impacted streams sufficient to meet key ecological
objectives? A Swedish case study. For. Ecol. Manag. 2021, 499, 119591. [CrossRef]
67. K˛edzior, R.; Kłonowska-Olejnik, M.; Dumnicka, E.; Woś, A.; Wyr˛ebek, M.; Ksia˛żek, L.; Grela, J.; Madej, P.; Skalski, T. Macroinver-
tebrate habitat requirements in rivers: Overestimation of environmental flow calculations in incised rivers. Hydrol. Earth Syst. Sci.
2022, 26, 4109–4124. [CrossRef]
68. Parreño, C.; Salazar Flores, C.; Slabospitskaya, A.S.; Adarchenko, I.A. Turbidity as a primary indicator of water quality from
surface water sources. Int. Res. J. 2024, 3, 1–7. [CrossRef]
Sustainability 2025, 17, 3684 32 of 32

69. Thirumalini, S.; Joseph, K. Correlation between Electrical Conductivity and Total Dissolved Solids in Natural Waters. Malays. J.
Sci. 2009, 28, 55–61. [CrossRef]
70. Scott, E.E.; Haggard, B.E. Natural Characteristics and Human Activity Influence Turbidity and Ion Concentrations in Streams. J.
Contemp. Water Res. Educ. 2021, 172, 34–49. [CrossRef]
71. Revelo-Mejía, I.A.; Gutiérrez-Idrobo, R.; López-Fernández, V.A.; López-Rosales, A.; Astaiza-Montenegro, F.C.; Garcés-Rengifo, L.;
López-Ordoñez, P.A.; Hardisson, A.; Rubio, C.; Gutiérrez, Á.J.; et al. Fluoride levels in river water from the volcanic regions of
Cauca (Colombia). Environ. Monit. Assess. 2022, 194, 327. [CrossRef] [PubMed]
72. Strauch, G.; Oyarzun, J.; Fiebig-Wittmaack, M.; González, E.; Weise, S.M. Contributions of the different water sources to the Elqui
river runoff (northern Chile) evaluated by H/O isotopes. Isot. Environ. Health Stud. 2006, 42, 303–322. [CrossRef] [PubMed]
73. Cedeño, D.V.G.; Maddela, N.R. Evaluación de calidad del agua de los ríos Canuto y Carrizal en época seca, Manabí, Ecuador.
Polo Conoc. 2022, 7, 537–554.
74. Abdallah, K.Z.; Hammam, G. Correlation between Biochemical Oxygen Demand and Chemical Oxygen Demand for Various
Wastewater Treatment Plants in Egypt to Obtain the Biodegradability Indices. Int. J. Sci. Basic Appl. Res. 2014, 13, 42–48.
75. Braga, F.H.R.; Dutra, M.L.S.; Lima, N.S.; Silva, G.M.; Miranda, R.C.M.; Firmo, W.C.A.; Moura, A.R.L.; Monteiro, A.S.; Silva,
L.C.N.; Silva, D.F.; et al. Study of the Influence of Physicochemical Parameters on the Water Quality Index (WQI) in the Maranhão
Amazon, Brazil. Water 2022, 14, 1546. [CrossRef]
76. Sotomayor, G.; Hampel, H.; Vázquez, R.; Goethals, P. Multivariate-statistics based selection of a benthic macroinvertebrate index
for assessing water quality in the Paute River basin (Ecuador). Ecol. Indic. 2020, 111, 106037. [CrossRef]
77. Nguyen, T.H.T.; Boets, P.; Lock, K.; Forio, M.A.E.; Van Echelpoel, W.; Van Butsel, J.; Dueñas Utreras, J.A.; Everaert, G.; Domínguez
Granda, L.E.; Hoang, T.H.T.; et al. Water quality related macroinvertebrate community responses to environmental gradients in
the Portoviejo River (Ecuador). Ann. Limnol. Int. J. Limnol. 2017, 53, 203–219. [CrossRef]
78. Banda, K.; Ngwenya, V.; Mulema, M.; Chomba, I.; Chomba, M.; Nyambe, I. Influence of water quality on benthic macroinverte-
brates in a groundwater-dependent wetland. Front. Water 2023, 5, 1177724. [CrossRef]
79. Simon, A.; Rinaldi, M. Disturbance, stream incision, and channel evolution: The roles of excess transport capacity and boundary
materials in controlling channel response. Geomorphology 2006, 79, 361–383. [CrossRef]
80. Santikari, V.P.; Murdoch, L.C. Effects of construction-related land use change on streamflow and sediment yield. J. Environ.
Manag. 2019, 252, 109605. [CrossRef]
81. Cashman, M.J.; Lee, G.; Staub, L.E.; Katoski, M.P.; Maloney, K.O. Physical habitat is more than a sediment issue: A multi-
dimensional habitat assessment indicates new approaches for river management. J. Environ. Manag. 2024, 371, 123139. [CrossRef]
[PubMed]
82. Haq, S.M.; Malik, Z.A.; Rahman, I.U. Quantification and characterization of vegetation and functional trait diversity of the
riparian zones in protected forest of Kashmir Himalaya, India. Nord. J. Bot. 2019, 37, e02438. [CrossRef]
83. Mosquera, G.M.; Hofstede, R.; Bremer, L.L.; Asbjornsen, H.; Carabajo-Hidalgo, A.; Célleri, R.; Crespo, P.; Esquivel-Hernández, G.;
Feyen, J.; Manosalvas, R.; et al. Frontiers in páramo water resources research: A multidisciplinary assessment. Sci. Total Environ.
2023, 892, 164373. [CrossRef]
84. Cortes, V.R.M.; Hughes, S.; Rodrigues Pereira, V.; Pinto Varandas, S. Tools for bioindicator assessment in rivers: The importance
of spatial scale, land use patterns and biotic integration. Ecol. Indic. 2013, 34, 460–477. [CrossRef]
85. Brumberg, H.; Beirne, C.; Broadbent, E.; Almeyda Zambrano, A.M.; Almeyda Zambrano, S.L.; Quispe Gil, C.A.; Lopez Gutierrez,
B.; Eplee, R.; Whitworth, A. Riparian buffer length is more influential than width on river water quality: A case study in southern
Costa Rica. J. Environ. Manag. 2021, 286, 112132. [CrossRef]

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