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Limiting factors for phytoplankton growth in

subtropical reservoirs: the effect of light and
nutrient availability in different longitudinal
compartments

Mr Davi Gasparini F. Cunha & Dr Maria do Carmo Calijuri

To cite this article: Mr Davi Gasparini F. Cunha & Dr Maria do Carmo Calijuri (2011) Limiting
factors for phytoplankton growth in subtropical reservoirs: the effect of light and nutrient availability
in different longitudinal compartments, Lake and Reservoir Management, 27:2, 162-172, DOI:
10.1080/07438141.2011.574974

To link to this article: https://doi.org/10.1080/07438141.2011.574974

Published online: 31 May 2011.

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Lake and Reservoir Management, 27:162–172, 2011

C Copyright by the North American Lake Management Society 2011
ISSN: 0743-8141 print / 1040-2381 online
DOI: 10.1080/07438141.2011.574974

Limiting factors for phytoplankton growth in subtropical

reservoirs: the effect of light and nutrient availability in different
longitudinal compartments

Davi Gasparini Fernandes Cunha∗ and Maria do Carmo Calijuri

School of Engineering in São Carlos, University of São Paulo (EESC-USP), Avenida
Trabalhador São-Carlense, 400, CEP 13566-590, São Carlos, SP, Brazil

Abstract
Cunha DGF, Calijuri MdC. 2011. Limiting factors for phytoplankton growth in subtropical reservoirs: the effect of
light and nutrient availability in different longitudinal compartments. Lake Reserv Manage. 27:162–172.

Limited information is available on the interactions between environmental factors and algal growth in tropical and
subtropical aquatic systems. We investigated the relationships between algal biomass (measured as chlorophyll,
Chl-a) and light, total phosphorus (TP) and total nitrogen (TN) in longitudinal zones of subtropical reservoirs.
We studied the seasonal variation of water variables in Itupararanga Reservoir (Brazil) and compared the results
with 16 other subtropical lakes and reservoirs. The longitudinal zones in Itupararanga Reservoir were considered
statistically different (p < 0.05, MANOVA). From the riverine zone to the dam region of the reservoir, Spearman
Correlation Test suggested that light limitation and TP limitation tended to decrease and increase, respectively.
Although nitrate concentrations were high (>400 µg/L), the Spearman coefficients between Chl-a and TN and the
TN:TP ratios (11:1 < TN:TP < 35:1) indicated that nitrogen may be co-limiting algal growth in the studied water
body. Putting Itupararanga in a regional context allowed assessment of potential influences of land use on trophic
state. Within the subtropical dataset, TP explained a greater percentage of variance in Chl-a (R2 = 0.70) than TN
(R2 = 0.17). The main land use type within the reservoirs drainage area significantly influenced the concentrations of
TP, TN, and Chl-a (p < 0.05, MANOVA), with different relationships between nutrients and chlorophyll in forested
(R2 = 0.12–0.33), agricultural (R2 = 0.50–0.68) and urban (R2 = 0.09–0.64) watersheds. Comparisons with literature
values and those from reservoirs with less altered watersheds indicated that Itupararanga Reservoir is reaching the
mesotrophic–eutrophic boundary, and further nutrient enrichment could cause water quality degradation.

Key words: light limitation, longitudinal zones, nutrient limitation, phytoplankton, subtropical reservoirs

Tropical and subtropical lakes have been less studied than
temperate lakes, and reservoirs less so than lakes. Thus,
there is limited information on factors influencing trophic
state and water quality in subtropical reservoirs. Thornton
et al. (1990) described reservoir compartmentalization,
where longitudinal gradients of nutrient and light availability
control chemical and biological gradients along the aquatic
system. The riverine zone is commonly characterized by
predominance of allochthonous organic matter as a result of
the contribution from the tributary rivers and by high turbid-
ity and low light availability. The transitional zone can have
optimum light and nutrients leading to high rates of primary
production. Nearer the dam, in the lacustrine zone, light
availability is high but nutrients can limit biomass accrual.
∗ scientific consensus about the primary limiting nutrient in
Corresponding author: davig@sc.usp.br
The compartmentalization concept proposed by Thornton
et al. (1990) presumes a particular reservoir configuration
that creates such nutrient and light gradients due to the river
influence.
Characterizing relationships among nutrients, light, and the
associated biomass response in aquatic systems, coupled
with watershed planning, allows a predictive approach to
lake management (Sterner and Grover 1998, Baird et al.
2001). Brazilian reservoirs are normally multipurpose sys-
tems (e.g., drinking supply, irrigation, and energy genera-
tion), and understanding their trophic characteristics is ben-
eficial for water quality and quantity control.
Nutrient effects on algal development differ both spatially
and temporally (Vincent et al. 1984). Currently, there is no

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 Limiting factors for algae in subtropical reservoirs
tropical–subtropical reservoirs, and specific spatial and tem-
poral patterns involved in nutrient–chlorophyll interactions
in such systems are not well characterized (Huszar et al.
2006). Most studies on controls of trophic state focused on
the relationships among total phosphorus, total nitrogen, and
chlorophyll a, primarily in temperate lakes and reservoirs,
although there were some efforts to model trophic state in
tropical reservoirs (e.g., Salas and Martino 1991, Huszar
et al. 1998). Models usually have nutrients driving standing
stock of chlorophyll (e.g., An and Park 2002, Freeman et al.
2009, Liu et al. 2010) to characterize the mechanisms in-
volved in biomass limitation (Håkanson et al. 2007, Becker
et al. 2010).
In this study, we aimed to investigate the roles of nutrients
and light in limiting biomass growth in subtropical aquatic
systems. We used the Itupararanga Reservoir, a long estab-
lished Brazilian reservoir (96 years), for detailed study and
compared the results with previously collected data from
16 other subtropical aquatic systems with urban-,
agricultural-, and forest-dominated watersheds. We hypoth-
esized that the limitation for algal growth is governed by
gradients of light and nutrient that occur along Itupararanga
Reservoir zones. We also expected higher total phospho-
rus and nitrogen concentrations in the studied reservoir in
comparison to presumably baseline values observed in for-
est dominated catchments (closest to pristine conditions).
Therefore, considered from a regional context, Itupararanga
was hypothesized to be at a more productive state than water
bodies with less altered watersheds.
Study site
Itupararanga Reservoir is a subtropical reservoir built in
1914, and its main use is for hydroelectricity generation
(historical mean discharge from 1914–2010 was 10.9 m3/s).
It is located in São Paulo State (Southeastern Brazil), in the
Sorocaba River Watershed (Fig. 1). The Una River, one of
the tributary rivers, was affected by domestic wastewater
discharges, with low dissolved oxygen concentrations and
high conductivity values (Salles et al. 2008).
Annual precipitation in the study area was about 1920 mm
in 2009, an exceptionally wet period in comparison to long-
term records; the historical average annual rainfall is around
1400 mm in the area (INMET 2010). Itupararanga Reservoir
has a storage capacity volume of about 286 million m3 and
an associated mean theoretical residence time from 4 to 13
months, depending on the rainfall pattern. Main land uses
in Itupararanga Reservoir catchment are agriculture (42%)
and native vegetation (25%; Garcia et al. 1999). In addition
to hydroelectricity generation, water from the reservoir was
used as a drinking water supply (2.2 m3/s for 800,000 people)
and irrigation (0.13 m3/s) in 2009.
Materials and methods
Four sampling sites were selected in the main channel of
Itupararanga Reservoir to cover the riverine (Riv), transi-
tional (Tra), lacustrine (Lac), and near-dam (Dam) zones
(Table 1; Fig. 1). Although the Dam station is also located
in the lacustrine zone, it is under the influence of the deep
water outlet for energy generation. Samples were collected
in August, October, and December 2009 and in February
2010 to be representative of distinct periods in terms of
rainfall, air temperature, and other climatological features.
We sampled preselected depths of the water column, al-
ways in the morning (between 8:00 and 10:00 h), with a van
Dorn sampler, corresponding to subsurface, 75, 50, 25, 1
and 0% (aphotic zone) of surface Photosynthetically Active
Radiation (PAR). Solar radiation in the water column was
measured with a radiometer (Quantameter Ly-Cor
R
). The
mixing zone depth (Zmix) was estimated as the depth of
the maximum thermal gradient through the assessment of
temperature profiles in the water column.
Total phosphorus (TP), total nitrogen (TN), total carbon
(TC), orthophosphate (PO4 i), nitrate (NO3 ), total suspended
solids (TSS), and nonvolatile suspended solids (NVS) con-
centrations were determined according to APHA (2005).
Chlorophyll a (Chl-a) was quantified according to Nush
(1980) following ethanol extraction. In situ measurements
for water temperature (T) and dissolved oxygen (DO) were
performed every 10 cm to a depth of 1.5 m and every 50 cm
for the remaining water column through a Multiparameter
Water Quality Probe (YSI 556
R
). Statistical manipulation of
the 11 studied water variables was performed through Mul-
tiway Analysis of Variance (MANOVA), using confidence
level of 95% (p < 0.05). MANOVA was used to verify the ex-
istence of significant differences among (1) the preselected
sampled depths in each station, (2) the longitudinal zones
(spatial heterogeneity), and (3) the sampling periods (tem-
poral variation). We also ran Principal Component Analysis
(PCA) for all the analyzed variables.
The 4 variables selected for Spearman’s Rank Correlation
Test were TP and TN (representing the nutrient role), Chl-
a (representing the biomass response), and Zmix (reflect-
ing the exposure of phytoplankton to the light within the
photoperiod). These variables were chosen because nutri-
ent and light availability control autotrophic metabolism
in water bodies (Biggs 2000, Neal et al. 2006). We used
mean concentrations for the water column for each sampling
site and period in this analysis because the sampled depths
were considered statistically similar (p > 0.4, MANOVA). A
significant Spearman correlation between nutrients and Chl-
a does not necessarily imply limitation by nitrogen or
phosphorus; therefore, absolute nutrient concentrations and
TN:TP were also analyzed and discussed in light of reference
values and limitation criteria available in the literature (e.g.,
163
 Cunha and Calijuri

Figure 1.-Location of the study area: Brazil, São Paulo State, major cities, Itupararanga Reservoir and the sampled sites in the riverine
(Riv), transitional (Tra), lacustrine (Lac), and near the dam (Dam) zones. Adapted from Marciano (2001).

Vollenweider 1968, Guilford and Hecky 2000, Dzialowski
et al. 2005, Haande et al. 2011).
We compiled TP, TN, and Chl-a data from 16 subtropical
reservoirs to put the results from Itupararanga Reservoir into
a regional context (Zmix data were not available). These
reservoirs (Table 2) varied in dominant land use within
their watersheds (CETESB 2009): urban (10 reservoirs:
Bambu e Do Meio, Barra Bonita, Billings, Garças, Graças,
Guarapiranga, Jaguari, Rio Grande, Taiaçupeba, and Tanque

164
 Limiting factors for algae in subtropical reservoirs

Table 1.-Sampling sites description with respective geographic coordinates and linear distance of each one from the station Riv.

Sites Description Geographic coordinates Linear distance from Riv (km)

Riv Riverine zone 23◦ 37 28 S; 47◦ 13 52 W —

Tra Transitional zone 23◦ 36 22 S; 47◦ 17 57 W 7.2
Lac Lacustrine zone 23◦ 36 50 S; 47◦ 20 11 W 10.6
Dam Near the dam 23◦ 36 55 S; 47◦ 23 25 W 16.2

Grande); agriculture (3 reservoirs: Cascata, Paranapanema,
and Promissão); and forest (3 reservoirs: Chavantes, Itaipu,
and Jurumirim). MANOVA was also used in this case to
verify if land use significantly influenced the concentrations
of TP, TN, and Chl-a. The dataset was obtained mainly from
CETESB (2007, 2008, 2009), an environmental agency of
São Paulo State (Brazil), but also from other publications
(Nogueira 2000, Calijuri et al. 2002, Crossetti 2006, Ribeiro-
Filho 2006, Lopes 2007, Gentil et al. 2008, Neves 2008).
Mean concentrations were calculated for TP, TN, and Chl-
a concentrations in a bimonthly basis when necessary. All
reservoirs or lakes are located in the southeast region of
Brazil, and the samples were taken from the deepest zone of
each aquatic system (lacustrine and dam regions). We used
linear regression to analyze relationships between nutrients
and chlorophyll.

Table 2.-Main characteristics (name, latitude, longitude, area, age, and years of collection) of the 16 lakes and reservoirs whose data were
compared to Itupararanga Reservoir results.

Geographic Area Age

Name coordinates (km2) (years) Years of data collection Reference

Bambu e Do Meio 23◦ 40 S 5 38 2005 and 2006 Lopes (2007)

Reservoir 46◦ 38 W
Barra Bonita 22◦ 32 S 310 47 1993, 1994 and 2007–2009 Calijuri et al. (2002); CETESB
Reservoir 48◦ 26 W (2007, 2008, 2009)
Billings 23◦ 43 S 127 52 2007–2009 CETESB (2007, 2008, 2009)
Reservoir 46◦ 39 W
Cascata 22◦ 12 S <1 38 2007–2009 CETESB (2007, 2008, 2009)
Reservoir 49◦ 55 W
Chavantes 23◦ 22 S 400 39 2005 and 2006 Neves (2008)
Reservoir 49◦ 36 W
Garças 23◦ 38 S <1 >100 1997–2004 Crossetti (2006); Gentil et al. (2008)
Lake 46◦ 36 W
Graças 23◦ 39 S <1 93 2007–2009 CETESB (2007, 2008, 2009)
Reservoir 46◦ 58 W
Guarapiranga 23◦ 45 S 27 >100 2007–2009 CETESB (2007, 2008, 2009)
Reservoir 46◦ 46 W
Itaipu 24◦ 05 S 1350 28 1999–2004 Ribeiro-Filho (2006)
Reservoir 54◦ 00 W
Jaguari 22◦ 55 S 56 29 2007–2009 CETESB (2007, 2008, 2009)
Reservoir 46◦ 25 W
Jurumirim 23◦ 15 S 449 48 1991 and 2007–2009 CETESB (2007, 2008, 2009);
Reservoir 49◦ 00 W Nogueira (2000)
Paranapanema 22◦ 39 S 576 32 2007–2009 CETESB (2007, 2008, 2009)
Reservoir 51◦ 23 W
Promissão 21◦ 17 S 530 35 2007–2009 CETESB (2007, 2008, 2009)
Reservoir 49◦ 46 W
Rio Grande 23◦ 46 S 7 29 2007–2009 CETESB (2007, 2008, 2009)
Reservoir 46◦ 32 W
Taiaçupeba 23◦ 34 S 20 34 2007–2009 CETESB (2007, 2008, 2009)
Reservoir 46◦ 17 W
Tanque Grande 23◦ 22 S <1 52 2007–2009 CETESB (2007, 2008, 2009)
Reservoir 46◦ 27 W

165
 Cunha and Calijuri

Table 3.-Main characteristics of Itupararanga Reservoir water column, including some physical, chemical, and biological variables (in
their mean values for all sampled depths) in Aug, Oct, and Dec 2009 and Feb 2010.

Period/ DO TSS NVS PO4 I NO3 TC Chl- Zmix TN TP

Station T (C) (mg/L) (mg/L) (mg/L) (µg/L) (mg/L) (mg/L) a(µg/L) (m) (µg/L) (µg/L) TN:TP

Aug 09 Riv 18.6 6.3 9.3 2.7 <10 0.84 10.4 31.8 0.6 935.8 72.7 13
Tra 17.6 7.1 3.7 0.6 <10 0.60 8.8 14.1 7.5 717.1 52.2 14
Lac 17.7 7.8 3.6 0.1 <10 0.44 8.8 15.6 10.5 543.5 46.5 12
Dam 17.9 8.6 3.7 0.1 <10 0.42 9.6 28.3 10.0 469.9 36.9 13
Oct 09 Riv 20.0 1.8 9.7 7.3 14.4 0.90 11.1 1.8 6.5 944.8 77.6 12
Tra 20.4 7.9 4.7 1.5 <10 0.55 9.6 17.5 2.0 613.6 22.9 27
Lac 20.3 7.4 2.9 0.9 <10 0.49 9.9 8.5 3.0 520.8 44.2 12
Dam 20.2 7.6 5.6 1.0 <10 0.45 8.9 14.6 5.0 486.4 21.8 22
Dec 09 Riv 21.4 1.4 7.8 4.4 13.2 0.82 12.4 7.3 7.5 879.1 77.8 11
Tra 23.2 5.7 4.6 1.2 <10 0.70 10.2 14.1 10.3 737.2 38.8 19
Lac 23.3 5.4 2.9 0.4 <10 0.60 9.3 15.5 10.0 638.1 29.9 21
Dam 23.4 6.5 4.3 1.0 <10 0.86 9.0 15.8 14.0 918.0 26.2 35
Feb 10 Riv 26.7 6.2 7.2 1.1 <10 0.67 12.7 24.9 1.5 814.5 73.2 11
Tra 26.2 3.5 4.2 0.6 <10 0.55 11.3 12.4 2.0 698.6 35.7 20
Lac 25.6 2.7 4.9 1.5 <10 0.55 11.1 12.1 3.5 636.6 48.5 13
Dam 25.1 3.5 3.8 1.1 <10 0.46 12.1 6.6 3.5 968.2 27.6 35

Results higher than 1000 µg/L for all cases. Water column phospho-
rus decreased from Riv (72.7–77.9 µg/L) to Dam (21.8–36.9
Itupararanga Reservoir µg/L), and TN:TP ratios varied from 11 to 35. The 4 ana-
We present mean values for all the variables in the water lyzed compartments in Itupararanga Reservoir were consid-
column because the preselected sampled depths were con- ered statistically different (p < 0.05, MANOVA) in relation
sidered statistically similar based on all studied variables to the 11 studied variables, highlighting the relevance of the
(p > 0.4, MANOVA). Itupararanga Reservoir water temper- spatial component.
ature showed significant variation depending on the sam- Among the 11 water variables assessed in Itupararanga
pling period (Table 3). The highest T of the water column Reservoir, the PCA selected only 5 abiotic variables
were detected in December and February (summer), reach- (Table 4), which were considered statistically significant
ing 26.7 C (Riv). The DO concentrations generally increased and explained 82.4% of data variability on the first two axes
from the riverine zone to the dam, except in February. In Oc- (Factor 1: 60.9% and Factor 2: 21.5%). The variables for
tober, for instance, DO concentrations ranged from 1.8 mg/L Factor 1 were PO4 i (0.94), TSS (0.93), NO3 (0.84), and TP
(Riv) to 7.6 mg/L (Dam) and in December from 1.4 mg/L (0.78). TN (0.99) was the only variable for ordination of
to 6.5 mg/L at the same locations. Factor 2. The PCA confirmed the close relation between
Suspended solids presented a decreasing trend from Riv to phosphorus (both TP and PO4 i) and TSS (Fig. 2).
Dam (from 7–9 mg/L to 3–6 mg/L). Higher concentrations
were found in the riverine zone for PO4 i (14.4 µg/L) and
NO3 (0.90 mg/L), with lower concentrations found in the
dam region (minimum of <10 µg/L and 0.42 mg/L, respec- Table 4.-Percentage of the total variance in the water variables in
Itupararanga Reservoir explained by principal components
tively). Total C presented the same pattern, with decreasing (Factor 1 and Factor 2) and the correlation values between Factor
concentrations from Riv (10.4–12.7 mg/L) to Dam (8.9–12.1 1 and 2 and the listed variables.
mg/L).
Factor 1 Factor 2
Chl-a concentrations (Table 3) were 1.8–31.8 µg/L (Riv),
12.4–17.5 µg/L (Tra), 8.5–15.6 µg/L (Lac), and 6.6–28.3 Variance explained 60.9% 21.5%
µg/L (Dam). Concentrations of NVS were higher in Riv PO4 i 0.938 0.139
TSS 0.926 0.013
(1.1–7.3 mg/L) than in the other sites (always lower than
NO3 0.842 −0.267
1.5 mg/L). Zmix presented considerable temporal variation,
TP 0.776 0.082
with the highest values observed in December (maximum TN 0.019 0.987
of 14 m, Dam). In comparison, TN concentrations were not

166
 Limiting factors for algae in subtropical reservoirs
Figure 2.-Plot of principal components (Factors 1 and 2). Vectors
plotted indicate the correlation scores for significant correlations
obtained between the water variables (in their mean values for all
sampled depths) and Factors 1 and 2. Numbers refer to the Riv,
Tra, Lac, and Dam stations in Aug (1, 2, 3, and 4, respectively), Oct
(5, 6, 7, and 8), Dec (9, 10, 11, and 12) and Feb (13, 14, 15, and
16).
The Spearman Correlation Coefficients between TP and
Chl-a in Itupararanga Reservoir varied from −0.8 in Riv
(indicating no limitation) to 0.4 in Dam, suggesting some
limitation (Fig. 3). The same coefficients between Zmix
and Chl-a increased from Riv (−0.8) to Dam (0.8), sug-
gesting light limitation decreased down the aquatic system.
The nitrogen limitation assessed by Spearman Coefficients
Figure 3.-Spearman Correlation Coefficients (ρ) between Zmix and
Chl-a (light limitation), TP and Chl-a (phosphorus limitation), and
TN and Chl-a (nitrogen limitation) for the sampling stations in
Itupararanga Reservoir (Riv, Tra, Lac, and Dam). Coefficients are
shown for significance correlation (p < 0.05).
presented higher coefficients in Riv and Lac (0.5 and 0.4,
respectively).
Other subtropical lakes and reservoirs
Mean TP, TN, and Chl-a concentrations (N = 202) for the 16
lakes and reservoirs included in the dataset were 62 µg/L,
1270 µg/L and 35 µg/L, respectively (Table 5). Most of
the data from the assessed subtropical lentic systems were
between 5 and 50 µg/L for TP, 23 and 1000 µg/L for TN,
and <1 and 25 µg/L for Chl-a. The scatterplots correlat-
ing log TP versus log Chl-a and log TN versus log Chl-a
(Fig. 4) showed that TP predicted more biomass variance
(R2 = 0.70) for the examined subtropical aquatic systems.
For phosphorus, the strongest correlation with Chl-a was
found in reservoirs located in agricultural watersheds (R2 =
0.68), and no relationship was observed in those aquatic
systems from areas with natural vegetation (Table 6). The
R2 values between TN and Chl-a were higher (0.50) for
reservoirs from agricultural catchments. Mean TP and TN
concentrations for reservoirs with forested watersheds were
15 µg/L and 500 µg/L respectively, and these levels could
be considered to represent the baseline trophic state for the
region.
Discussion
The sampling sites and the collection periods were con-
sidered statistically different based on all studied physical,
chemical, and biological variables in Itupararanga Reservoir
(p < 0.05, MANOVA). The DO patterns from Riv to Dam
were probably linked with the algal primary production and
with the decrease of oxygen demand that is supposed to oc-
cur from the headwater of the reservoir to the dam. Causes
could be suspended material sedimentation, organic matter
assimilation and mineralization by biological communities,
or both. The variation of TSS, NVS, and TC concentra-
tions longitudinally along the reservoir also indicated this
potential relationship (i.e., higher organic matter and solids
Table 5.-Mean, minimum, and maximum TN:TP ratios and
concentrations for TP, TN, and Chl-a comprising the dataset from
16 subtropical lakes and reservoirs. The number of data points
(N) and the standard deviation (s.d.) are also shown for each
case.
Variable N Mean s.d. Min. Max.
TP (µg/L) 202 62 67 5 334
TN (µg/L) 202 1270 1838 23 11,559
Chl-a (µg/L) 202 35 72 <1 855
TN:TP 202 36 41 <1 204
167
 Cunha and Calijuri

Table 6.-TP, TN, and Chl-a relationships considered individually for reservoirs with different land use in their watersheds (urban,
agricultural, and forested). The number of available data (N); the mean, minimum (min), and maximum (max) concentrations; the
strength of the correlation (as R2); and the respective regression equation (when R2 ≥ 0.50) are shown for each case.

Catchment main Min-Max TP

land use N Mean TP (µg/L) (µg/L) TP vs. Chl-a R2 TP vs. Chl-a Equation

Urban 135 79 5–334 0.64 log chl a = −0.496 + 1.062 log TP (1)
Agricultural 21 55 10–260 0.68 log chl a = −0.921 + 1.395 log TP (2)
Forest 46 15 6–36 0.33 —

Catchment main Min-Max TN

land use N Mean TN (µg/L) (µg/L) TN vs. Chl-a R2 TN vs. Chl-a Equation

Urban 135 1487 23–11, 559 0.09 —

Agricultural 21 1564 250–9780 0.50 log chl a = −2.919 + 1.358 log TN (3)
Forest 46 500 58–1910 0.12 —

content in the riverine zone and lower near the dam). Chl-a
concentrations presented larger variation in Riv, indicating
the possibility of allochthonous contribution for this vari-
able (e.g., chlorophyll from the tributary rivers). The PCA
confirmed the strong effect of allochthonous organic matter
in the riverine portion of the reservoir. The plot of princi-
pal components clearly showed the influence of phosphorus
and solids for separating Riv from other sampling sites in
the studied periods (Fig. 2).
Spearman correlation coefficients were used as auxiliary
indicators of factors limiting phytoplankton growth. The
coefficients between Zmix and Chl-a suggested that light
limitation decreased from the riverine zone to the dam of
Itupararanga Reservoir. The possible light limitation de-
crease from the headwaters to the lacustrine compartment
is compatible with TSS and NVS results and consistent
with the expected longitudinal flow dynamics. From Riv,
the sedimentation of suspended particles increases as wa-
ter flux decreases. Figueiredo and Bianchini (2008) also
suggested that the hydraulic factor (sedimentation) could
improve water transparency in a Brazilian reservoir. Light
availability in the lacustrine zone is frequently recognized
as an important factor for stimulating phytoplankton density
increase (Molisani et al. 2010). Moreover, low Zeu/Zmix ra-
tios are normally associated with poor biomass development
(Borges et al. 2008) because Zeu is restricted and Zmix is
high, thus imposing dark conditions for algae during part of
the photoperiod.
Internal nitrogen losses are common in tropical aquatic
systems as a result of higher water temperatures (Lewis
2000, 2002), which could lead to limitation by this
nutrient. Nevertheless, atmospheric fixation continuously

Figure 4.-Relationships between total phosphorus (as log TP) and total nitrogen (as log TN) vs. chlorophyll a (as log Chl-a) for 16
subtropical reservoirs. The number of available data (N), the regression equation, and the significance of the correlation (as R2) are shown
for each case. Data from Itupararanga Reservoir were also plotted in the graphs.

168
 Limiting factors for algae in subtropical reservoirs
contributes to nitrogen replacement. In addition, cyanobac-
teria are of widespread occurrence in Brazilian waterbodies
and are able to fix nitrogen through the heterocysts (Sprõber
et al. 2003, Fiore et al. 2005, Figueredo and Giani 2009,
Haande et al. 2011). For these reasons, phosphorus limita-
tion is frequently reported in tropical and subtropical reser-
voirs (Gianesella-Galvão 1985, Páez 2001). In recent years,
however, a limitation chain with N, P, and light was pro-
posed for phytoplankton. The transition between N and P
limitation is no longer considered a sharply established point
(Pahlow and Oschlies 2009).
Spearman coefficients (ρ) between Chl-a and TN (maximum
of 0.5) indicating some nitrogen limitation were observed
especially in Riv and Lac of Itupararanga Reservoir. Some
recent research reported nitrogen limitation when NO3 con-
centrations are low, for example <10 µg/L in Lake Victoria,
Uganda (Haande et al. 2011). In the case of Itupararanga
Reservoir, NO3 concentrations in all sampled sites were al-
ways higher than 400 µg/L, and TN:TP ratios were never
below 11:1. Although some authors considered the possibil-
ity of no nitrogen limitation when TN:TP ratios are greater
than 9:1(Vollenweider 1968), N and P co-limitation has re-
cently been described as much more common than previ-
ously thought (Elser et al. 2007).
Criteria presented in the literature for N and P co-limitation
are based on TN:TP ratios, 20 < TN:TP < 50 (Guilford
and Hecky 2000) or 20 < TN:TP < 46 (Dzialowski et al.
2005). Therefore, considering that TN:TP ratios varied be-
tween 11 and 35 in Itupararanga Reservoir (Table 3) and
that the Spearman Coefficients were not high for either N
or P, we considered the possibility of nutrient co-limitation
in Itupararanga. The variation of the Spearman Coefficients
across different longitudinal zones is also another indica-
tion of the existence of multiple factors (nutrients and light)
co-regulating algal development.
The comparison of the results in Itupararanga Reservoir with
16 other systems (Fig. 4) showed that TP explained more
of the total variance for Chl-a (70%) in comparison to TN
(17%). However, mean TN:TP was 36 for the dataset, and N
and P co-regulation of algal growth may also have occurred
in 40% of cases for which 20 < TN:TP < 46, the criteria
established by Dzialowski et al. (2005). Land use signif-
icantly influenced nutrient and chlorophyll concentrations
(p < 0.05, MANOVA). The percentage of Chl-a variance
explained by TN was greater for reservoirs with agricultural
catchments (50%; Table 6).
We used the 95% confidence interval (Kleinbaum and Kup-
per 1978) around the slope of our regression equation with
TP versus Chl-a (Fig. 4) to make a comparison with the slope
of another equation reported in the literature. Our regression
equation was significantly different from that obtained by
Huszar et al. (2006), who studied 192 tropical and subtropi-
cal lakes (R2 = 0.42). The slope of our equation was greater,
indicating a more significant response of chlorophyll to TP
in Brazilian lakes and reservoirs than verified by Huszar et
al. (2006) for their dataset.
TP concentrations explained 68% of Chl-a variance when
we only considered the reservoirs located in agricultural
areas (Table 6; N = 21). In contrast, poor correlation was
observed between log TP and log Chl-a for those reservoirs
located in forested watersheds, what may be associated with
the lower TP concentrations (always <36 µg/L).
The small TP and TN variability in the Itupararanga Reser-
voir relative to the entire datasets and the aforementioned
possibility of co-limitation by P and N could partly ex-
plain why correlation between each nutrient and chlorophyll
was relatively weak in Itupararanga. Itupararanga Reservoir
could be reaching the boundary between a mesotrophic and
eutrophic state, considering the trophic state classification
presented by Lamparelli (2004), who proposed a trophic
state index for tropical reservoirs, calculated through TP
and Chl-a concentrations.
TP and TN concentrations observed in Itupararanga were
respectively up to 2 and 5 times higher than the average
concentration for reference reservoirs located in forested
watersheds (i.e., the least impacted regional reservoirs),
whose mean TP and TN concentrations were 15 µg/L
and 500 µg/L (Table 6). Such a comparison reinforced
the importance of establishing nutrient criteria to avoid
eutrophication and negative effects on water uses of lentic
and lotic systems (Dodds and Welch 2000, Smith et al.
2003, Dodds and Oakes 2004). Our average reference con-
centrations for TP and TN are similar to those described by
Lamparelli (2004), who presented baseline concentrations
for São Paulo state (Brazil) reservoirs: 15–30 µg/L (TP)
and 810–840 µg/L (TN).
Finally, the Chl-a concentration associated with the mean
TP concentration in Itupararanga Reservoir during our sam-
pling period (52 µg/L) was 30.0 µg/L, according to equation
2 (Table 6) for agricultural catchments. This is exactly the
upper limit imposed by the Brazilian legislation for Chl-
a concentrations in Itupararanga Reservoir (Brasil 2005).
Therefore, if nutrients levels increased, the aquatic system
could be affected by the undesirable effects of artificial eu-
trophication (e.g., algal blooms, taste and odor problems,
health risks, and decrease in property values), in addition
to potential conflicts with legislation. These findings are
consistent with recent reports from the São Paulo State En-
vironmental Company (CETESB 2007, 2008, 2009) that
stressed the increasing trophic state of Itupararanga Reser-
voir in recent years and the need for adequate management
for recovering its water quality.
169
 Cunha and Calijuri
Conclusions
The results obtained for Itupararanga Reservoir, consider-
ing the restrictions related to data availability and the use of
auxiliary tools (statistic manipulation and criteria available
in the literature), suggested that phosphorus and nitrogen
co-limitation is more likely to occur in this aquatic system.
Light and nutrient limitation seemed to play an alternating
role within its different longitudinal zones. Light conditions
were probably able to restrict algae growth in the riverine
zone, and their importance seemingly decreased as solids
settled and water transparency increased in the other re-
gions. TP limitation probably did not occur in the riverine
zone of Itupararanga Reservoir, as a result of allochthonous
phosphorus from riverine inputs; however, the strength of
nitrogen limitation was probably greater in the riverine and
lacustrine zones, as suggested by the Spearman Coefficients.
Regarding the other 16 analyzed subtropical lakes and reser-
voirs, the results indicated that phosphorus explained more
of the chlorophyll variance in relation to nitrogen; however,
TP and TN co-limitation may also have occurred, based on
the TN:TP ratios. Land use within the reservoir drainage
area significantly influenced the nutrients and chlorophyll
concentrations, although further studies are necessary (espe-
cially in agricultural catchments because the number of data
points for this case was limited to 21). Therefore, the estab-
lishment of nutrient criteria for Brazilian reservoirs should
necessarily consider these differences in relation to water-
shed land use to avoid eutrophic conditions. In the wider
context of regional reservoirs, TP and TN concentrations in
Itupararanga Reservoir were, respectively, 3.1 and 1.4 times
higher in average terms than reference (i.e., more pristine)
conditions. Considering that the aquatic system seems to be
transitioning from mesotrophic to eutrophic status, further
nutrient enrichment could contribute to water quality deteri-
oration, having a negative effect on the multiple uses of the
reservoir (especially drinking water supply and recreation).
Future research within tropical–subtropical reservoirs
should perform complementary studies (e.g., phytoplank-
ton community investigation and nitrogen fixation measure-
ments) to strengthen our conclusions on limitation factors.
This would certainly favor the exploration of the described
mechanisms for other aquatic systems and aid in trophic
state assessment through the use of nutrient concentrations,
light conditions, and land use information.
Acknowledgments
The authors are grateful to FAPESP (Fundação de Amparo à
Pesquisa do Estado de São Paulo) for the scholarship to DGF.
Cunha (Process Number 2009/50842–2) and for the finan-
cial support to MC. Calijuri (Process Number 2008/55636-
170
9). Also, the authors express their sincere thanks to Walter K.
Dodds, Danelle Russell, to the members of the Journal Club
(Kansas State University), and to 3 anonymous reviewers
for valuable suggestions for improving the manuscript.
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