Lipids

DOI 10.1007/s11745-016-4196-z

ORIGINAL ARTICLE

Fatty Acid Composition ofTropical Fish Depends onReservoir

Trophic Status andFish Feeding Habit
AlineD.Gomes1 CarlosE.Tolussi1 IolaG.Bochat2 MarceloL.M.Pompo3
MaraP.T.Cortez3 RenatoM.Honji1 RenataG.Moreira1

Received: 6 July 2016 / Accepted: 27 August 2016

AOCS 2016

Abstract Eutrophication results in a deficiency of n-3 LCPUFA (long-chain polyunsaturated fatty acids) in aquatic
food chains, affecting fish nutrition and physiology. The
trophic transfer of FA (fatty acids) to fish species of different feeding habits was investigated in two reservoirs in
southeast Brazilthe mesotrophic Ponte Nova Reservoir
(PN) and the hypereutrophic Billings Reservoir (Bil). Total
FA profile of stomach contents and adipose tissue, triacylglycerols (TAG), and phospholipids (PL) from liver and
muscle of the omnivorous Astyanax fasciatus and the carnivorous Hoplias malabaricus were analyzed by gas chromatography. A prevalence of n-6PUFA, as 18:2n-6 (linoleic
acid) and 20:4n-6 (arachidonic acid, ARA) was observed in
the stomach contents and in the tissues of A. fasciatus from
the PN reservoir. In contrast, n-3 LC-PUFA, as 20:5n-3
(eicosapentaenoic acid, EPA) was accumulated in fish tissues from Bil, resulting in higher n3/n6 and EPA/ARA
ratios, compared to fish from PN. This differential FA accumulation was also observed for H. malabaricus, but differences were slightly minor, and no changes were observed
in the EPA/ARA ratios between fish from both reservoirs.
Regardless reservoir, FA profiles of TAG resembled that of
their diet, whereas FA profiles of PL were more conservative and mainly comprised by LC-PUFA. We conclude that

* Aline D. Gomes
eniladal@gmail.com
1

Departamento de Fisiologia do Instituto de Biocincias,

Universidade de So Paulo, Rua do Mato, Trav. 14, 321, So
Paulo, SP 05508090, Brazil

Departamento de Geocincias, Universidade Federal de So

Joo Del Rei, So Joo Del Rei, MG, Brazil

Departamento de Ecologia do Instituto de Biocincias,

Universidade de So Paulo, So Paulo, SP, Brazil

reservoir trophic status affected the FA composition of food

resources available to these fish species, resulting in differential allocation of n-3 and n-6 FA. As expected, FA profile
of the investigated fish species also reflected their feeding
habit and physiological demands.
Keywords Eutrophication Feeding habit Food chain
Polyunsaturated fatty acids
Abbreviations
18-PUFA Polyunsaturated fatty acid with 18
carbons
LC-PUFA Long-chain polyunsaturated fatty acid
(with 20-22 carbons)
Af 
Astyanax fasciatus
ARA (20:4n-6) Arachidonic acid
AT Adipose tissue
BFA Branched chain fatty acids
Bil Billings (hypereutrophic reservoir)
DHA (22:6n-3) Docosahexaenoic acid
EPA (20:5n-3) Eicosapentaenoic acid
FA Fatty acids
FAME Fatty acid metil-esters
Hm 
Hoplias malabaricus
HPL Hepatic phospholipids
HTAG Hepatic triacylglycerols
MPL Muscle phospholipids
MTAG Muscle triacylglycerols
MUFA Monounsaturated fatty acids
n-3 Omega 3
n-6 Omega 6
OFA Odd chain fatty acids
PL Phospholipids
PN Ponte Nova (mesotrophic reservoir)
PUFA Polyunsaturated fatty acids

13

 Lipids

SC Stomach content
SDA Stepwise discriminant analyses
SFA Saturated fatty acids
TAG Triacylglycerol
TSI Trophic state index

Introduction
In freshwater aquatic systems, the polyunsaturated fatty
acids with 18 carbons (18-PUFA) 18:3n-3 (-linolenic
acid) and 18:2n-6 (linoleic acid) are essential to all animals,
and must be obtained from their diet [14]. Primary producers synthesize PUFA, which are transferred throughout
the food chain up to consumers [5]. Green algae and cyanobacteria produce high amounts of 18:3n-3 and 18:2n-6
[6]. Diatoms and cryptophytes produce n-3 LC-PUFA
(long-chain fatty acids), such as 20:5n-3 (eicosapentaenoic
acid, EPA) and 22:6n-3 (docosahexaenoic acid, DHA) [7,
8]. However, fatty acid (FA) composition in consumer tissues may differ from that of their food resources, due to
biochemical processes, including selective mobilization of
FA, catabolism via -oxidation, and de novo synthesis [9].
For example, after the incorporation of 18-PUFA in their
tissues, animals can convert 18:3n-3 to EPA and DHA and
18:2n-6 to 20:4n-6 (arachidonic acid, ARA). Conversion
efficiency, though, differs among species [10].
LC-PUFA of the n3 and n6 families are physiologically
very important, as they are crucial for growth, reproduction,
and development of the immune system in vertebrates and
invertebrates [1114]. DHA is especially abundant in fish
retina and brain, and plays an important role in the structure and function of cellular membranes [1518]. EPA and
ARA are precursors of eicosanoids, compounds that act as
local hormones or signaling molecules involved in inflammation response, immunity [13], and reproduction [1921].
Moreover, n-3 and n-6 derived eicosanoids often compete
for enzymes in the pathway synthesis and act antagonistically in several physiological processes. Hence, the EPA/
ARA ratio in tissues can determine the action of eicosanoids in fish physiology [22].
The FA profile of fish varies in response to many factors, including the physiological status and dietary availability, which may be affected by environmental conditions.
A study carried out in the Great Lakes, Canada, showed
that populations of the amphipod Diporeia spp. decreased
in many areas due to the introduction of invasive mussels,
mainly Dreissena bugensis [23]. Diporeia spp. are rich in
EPA and DHA, and their decline in the environment seriously affected the availability of n-3 LC-PUFA to local fish
species [13]. Cultural eutrophication process, the excessive algae growth due to nutrients enrichment from human
activities [24], may also affect the basis of the food web,

13

with bottom-up consequences for the entire food web.

Mller-Navarra et al. [24] observed in Canadian eutrophic
lakes, a replacement of taxa rich in n-3 LC-PUFA (mainly
EPA and DHA), such as diatoms, cryptophytes and dinoflagellates, with n-3 PUFA-poor taxa, such as chlorophytes and cyanobacteria. As a consequence, zooplankton
experienced lower somatic growth and reproduction rates.
Moreover, by affecting community composition and the
nutritional status of available resources, eutrophication is
likely to affect the trophic transfer of important biochemical precursors and other essential LC-PUFA. Such changes
in the nutritional quality of lower trophic levels may result
in FA limitation in fish. FA limitation ultimately affects cellular membrane fluidity and eicosanoid synthesis [13], but
effects may differ depending on the main food resource and
the fish species.
Most studies involving trophic transfer of PUFA have
been carried out primarily on phytoplanktonherbivorous
zooplankton interface, and in temperate lakes [2527].
Studies focusing on freshwater fish are less common
[2831] and virtually absent in regions of tropical climate.
We hypothesized that fish species from a hypereutrophic
tropical reservoir, highly enriched with nutrients and algae
growth, will display a deficiency of n-3 LC-PUFA in their
tissues, compared to fish species from a mesotrophic system, with low nutrients and algae growth, as a result of
dietary FA deficiencies generally found in hypereutrophic
reservoirs. However, the effects should be smoother for
carnivorous than for omnivorous fish. Omnivorous species
would partially rely on n-3-poor phytoplankton as a food
resource in such hypereutrophic systems, and thus obtain
n-3-poor food in larger amounts than carnivorous fish
would. Carnivorous fish are likely to receive n-3 LC-PUFA
synthesized by their animal prey, by feeding on zooplankton and other smaller fish. Therefore, carnivorous fish will
be less subjected to the low quality of phytoplankton than
omnivorous fish in hypereutrophic reservoirs. To test these
hypotheses, females of two fish species, one omnivorous
and one carnivorous, were selected from two reservoirs of
different trophic status in Brazil. The FA profile of different tissues (adipose, liver, and muscle) was compared to
that of the stomach content (as a proxy for the FA profiles
of food resources) of both fish species within and between
reservoirs.

Materials andMethods
Study Site
The present study was carried out in two reservoirs of
the Tiet River sub-basin (So Paulo State, southeastern
Brazil) (Fig.1a, b; Table1). Further limnological details

Lipids
Fig. 1a, b Map of the Tiet
Sub-Basin, So Paulo State
(Brazil), showing the location of sampling sites: c The
Ponte Nova Reservoir (PN;
mesotrophic reservoir), and
d the Billings Reservoir (Bil;
hypereutrophic) highlighting the
Taquacetuba branch

Table1Hydrogeomorphic,
physical, chemical, and
biological characterization
of the mesotrophic (Ponte
Nova, PN) and hypereutrophic
reservoirs (Billings, Bil)

Variables

PN

Bil

Surface area (km2)

Water retention time (days)
Elevation (m)
Depth of the Secchi disk (m)
Temperature (C)
Dissolved oxygen (mg L1)
pH

25.7
720
840
1.44.85
1623
68
6.5
0.74.1

106.6
80
745
0.72.5
1725.5
7.510
7.59.5
33.3867.0

9.5200
2.09.0
500.00900.0
8.020.0
Mesotrophic
Green algae, flagellates,
diatoms

288.91045.9
8.125.7
430.0473.6
54.6402.2
Superhypereutrophic

[3235]

[3537]

Chorophyll-a (g L1)
Nitrate (g L1)
Nitrite (g L1)
Total nitrogen (g L1)
Total phosphorus (g L1)
TSI
Dominant phytoplanktonic
groups
References

Cyanobacteria (constant bloom),

dinoflagellates and green algae

TSI Trophic State Index

about these reservoirs are available in Gomes et al. [35].

The mesotrophic Ponte Nova Reservoir (PN) is located in
a watershed area under environmental protection, far from
urban centers and industries. This region is occupied by
Atlantic Forest and preserved riparian vegetation and lowland along the waterway [35, 38]. Sampling was carried
out in Rio Claro branch (PN; 233436.5S; 455423.9W;
Fig. 1c). The hypereutrophic Billings Reservoir (Bil) is
characterized by intense anthropogenic impacts, such

as discharge of domestic sewage and untreated industrial effluents, riparian deforestation, and extensive land
use. Sampling was carried out in the Taquacetuba branch
(234816.5S; 463833.26W; Fig.1d), which is considered hypereutrophic due to high urbanization and occasional cyanobacterial blooms [35, 36, 39]. Moreover, high
concentrations of heavy metals have been found in this
reservoir, such as cadmium, lead, copper, mercury, and
nickel [35].

13

 Lipids

Data Collection andProcessing

Astyanax fasciatus and Hoplias malabaricus were selected
for this study because they are among the most frequently
caught fish in the studied reservoirs. A. fasciatus is a small
fish with a geographical distribution throughout Central
and South America [38] and has an omnivorous feeding
habit, including mainly zooplankton and insect prey organisms [40, 41]. H. malabaricus is a neotropical fish species,
essentially piscivore, with a wide geographic distribution
in South America, including nearly all hydrographic watersheds, except the transandine areas and Patagonian rivers
[4244].
Throughout the year 2012 (summer: DecemberMarch;
autumn: MarchJune; winter: JuneSeptember; spring:
SeptemberDecember), were sampled fish from both reservoirs, with a total of 82 adult females of the omnivorous
fish A. fasciatus (body mass: 15.24.09g; total length:
10.3 0.76cm) and 53 adult females of the carnivorous
fish H. malabaricus (body mass: 647.7135.23g; total
length: 36.94.77cm). Fish were captured using gillnets
placed for 12h on the water surface. Individuals captured
alive were kept in plastic containers with water from the
reservoir until dissection, which was done on the same day
of capture. In the laboratory, the individuals were anesthetized with 1g of benzocaine (ethyl-p-aminobenzoate) dissolved in 10mL ethanol and 10 L of water. Morphometric
and weight data were recorded and the animals were euthanized by sectioning their spinal cord, as approved by the
Animal Ethics Committee of the Institute of Biosciences,
University of So Paulo (Protocol number 154/2012).
Finally, samples of adipose tissue, liver and muscle were
removed and stored at 80C until further analysis.
Stomachs were removed intact from the captured fish
and scraped in order to collect the content, also stored at
80C until analysis.
Fatty acid analysis
Total lipids of stomach contents and tissues were extracted
using chloroform/methanol/water (2:1:0.5, v:v:v) [45, 46].
Total lipids extracted from liver and muscle were fractionated in triacylglycerol (TAG) and phospholipid (PL) fractions using thin layer chromatography [47]. TAG and PL
fatty acids of liver and muscle, as well as total lipid fatty
acids of stomach contents and adipose tissues, were methylated with acetyl chloride (5% HCl in methanol) and converted into FA methyl-esters (FAME) [48].
FA analysis was carried out in a Varian gas chromatograph (GC; Model 3900, Walnut Creek, CA, USA) coupled with a flame ionization detector (FID) and a CP-8410
autosampler. FAMEs were analyzed on a capillary column (CP Wax 52 CB, 0.25m thickness, 0.25mm inner

13

diameter, and 30m length). Hydrogen was used as the carrier gas at a linear velocity of 22cm/s. The column was
programmed at 170C for 1min, followed by a 2.5C/min
ramp to 240C and a final hold time of 5min. The injector
and FID temperatures were 250 and 260C, respectively.
FAME were identified by comparing their retention times
to those obtained from commercial standards (Supelco, 37
components; SigmaAldrich; Mixture, Me93, Larodan and
Qualmix, PUFA fish M, Menhaden Oil, Larodan). Data are
presented as mole% of total FAME based on peak areas
analyses.
Data Analysis
Since PUFA composition of stomach contents and tissues
TAG and PL from both species and reservoirs showed no
significant temporal variation, data from all sampled seasons were pooled together. Differences in the FA profile of
each fish tissue between reservoirs and species were tested
by the Student t test. The significance level adopted was
95% (p<0.05). The statistical analysis was carried out
using SigmaPlot for Windows 10.0 (Systat Software Inc.,
San Jose, CA, USA).
Stepwise discriminant analyses (SDA) were performed
to determine which combination of variables (FA) better
discriminates between reservoirs and species. A total of
37 FA were identified in the samples, however, to meet the
assumptions of sample size and variables of the SDA, eight
FA were selected (16:0, 18:0, 18:1n-9, 18:2n-6, 18:3n-3,
20:4n-6, 20:5n-3, and 22:6n-3), which were abundant in all
samples and for which statistical differences were previously detected by a Student t test. To search for differences
caused by the type of system (hypereutrophic and mesotrophic), a separate SDA was performed for each species.
As FAs have different physiological functions depending
on the molecule they belong to, lipid classes were separated
and analyzed two separate SDA per species, with either PL
or TAG fatty acids. Finally, a SDA with TAG fatty acids
was made using both fish species, in order to access group
discrimination related to species-specific differences in FA
profiles and discrimination related to the environmental
conditions in both reservoirs. In this analysis, only TAG
fatty acids were used, as they better reflect the diet profile
[49]. Statistical analyses were carried out with IBM SPSS
Statistics for Windows, version 20 (IBM Corp., Amonk,
NY, USA).

Results
Stomach contents of A. fasciatus from the PN reservoir, as
well as tissue TAG samples, showed higher percentages of
MUFA (monounsaturated fatty acids), mainly 18:1n-9, and

Lipids

n-6 PUFA, mainly 18:2n-6, than those from Bil reservoir,

which were richer in n-3 PUFA (including 18:3n-3, EPA,
and DHA) (p<0.001 in all cases; Table2). This same
PUFA difference was observed in PL samples; however,
the higher values of n-6 PUFA from females captured in
the PN reservoir were mainly due to ARA, while elevated
percentages of n-3 PUFA in the PL of fish tissues from the
Bil reservoir were due to 18:3n-3 and EPA (p<0.001 in
all cases; Table3). The observed differences among PUFA
profiles of TAG and PL resulted in higher n3/n6 and EPA/
ARA ratios in female fish from the Bil reservoir (p<0.001;
Tables2, 3).
Only EPA percentages were higher in the stomach content of H. malabaricus from Bil reservoir, compared to the
same species captured from PN reservoir (p =0.034). As
a result, there were also higher n-3 PUFA percentages and
n3/n6 ratios in these animals (Table4). Similarly, single
differences observed in the FA profile of tissues between
reservoirs were also only related to PUFA. Thus, TAG and
PL of females from PN reservoir showed higher percentages of n-6 PUFA, such as 18:2n-6 and ARA, than TAG
and PL from females from the Bil reservoir, which had, in
turn, higher percentages of n-3 PUFA, such as 18:3n-3 and
EPA (p<0.05, Tables3, 4). The n3/n6 ratio was also higher
in tissues of females from the Bil reservoir (Tables3, 4).
However, the EPA/ARA ratio in PL did not differ between
reservoirs (Table3).
In all SDA, variables within the first function explained
more than 50% of the total variation among groups
(Table5), and all analyses resulted in two significant functions (p<0.001). The first SDA, run with data of TAG fatty
acid composition of A. fasciatus, provided the best discrimination between groups (Fig.2a). The first function separated samples according to the reservoir, due to differences
in 18:3n-3 and 18:2n-6 percentages (Table5; Fig.2a). The
second function separated FA composition of stomach content and adipose tissue from the FA composition of hepatic
and muscle TAG, based on differences in 18:0, 18:2n-6,
ARA, and EPA percentages (Table5; Fig.2a). The second
SDA, run with PL data of A. fasciatus tissues, revealed a
first function that separated FA composition of the stomach content from all other samples analyzed, based on
differences in 18:1n-9 and 18:2n-6 percentages (Table5;
Fig. 2b). The second function separated samples between
reservoirs, according to differences in 18:1n-9, 18:2n-6,
EPA, and ARA percentages (Table5; Fig.2b).
The SDA based on TAG fatty acids of H. malabaricus
tissues provided the best separation between reservoirs in
the first function, based on differences in 18:3n-3, ARA,
and EPA percentages. The second function separated FA
profiles of muscle and adipose tissue, mainly from liver,
regardless of the reservoir. Differences were based on
ARA, EPA, and DHA percentages (Table5; Fig.3a). The

SDA based on PL fatty acids showed the separation of

muscle from all tissues analyzed in the first function, due
to 18:2n-6, EPA, and DHA percentages (Fig.3b). The second function separated FA profiles of stomach contents and
muscle of animals from the hypereutrophic reservoir from
all other samples, based on differences in 16:0 and ARA
(Table5; Fig.3b). Finally, a third SDA, based on TAG fatty
acids of both species together, separated reservoirs regardless of species and tissues, based on differences in 18:3n-3
and 18:2n-6 percentages (Table5; Fig.4a). The second
function separated H. malabaricus from A. fasciatus from
both reservoirs, according to differences in EPA percentages (Table5; Fig.4a).
Concerning differences between species, the percentage
of 18-PUFA (18:3n-3 and 18:2n-6) decreased from omnivorous to carnivorous species in both reservoirs (p<0.001;
Fig. 4b), while LC-PUFA increased (p<0.001; Fig.4b).
Additionally, differences in n-3/n-6 and EPA/ARA ratios
of all analyzed tissues between reservoirs were always
higher for the omnivorous species (39 times higher for n3/
n6 and 37 times higher for EPA/ARA ratios in Bil than in
PN samples) than for the carnivorous species (0.53 times
higher for n3/n6 and, 23 times higher for EPA/ARA ratios
in Bil than in PN samples) (Tables2, 3, 4).

Discussion
Fatty acids differed markedly between fish species and reservoirs, and those differences varied depending on the tissue. The higher percentages of n-3 PUFA, including EPA
and DHA, in stomach contents and tissues of both species from the hypereutrophic reservoir (Bil) suggest that
food resources in this reservoir were not as deficient in
n-3 LC-PUFA as initially expected. Because of the occurrence of cyanobacterial blooms in the Bil reservoir [35,
36] we expected to find higher percentages of 18:2n-6 and
18:3n-3 in the food resources, since these are the main FA
produced by cyanobacteria [7]. Instead, besides high percentages of 18:3n-3, we also found high percentages of
n-3 LC-PUFA, mainly EPA in the stomach contents of fish
captured from the hypereutrophic reservoir, which contradicts the expected pattern for this reservoir [24]. In recent
years, a high density of dinoflagellates has been identified
in the Bil reservoir, concomitantly with the occurrence of
cyanobacterial blooms [50, 51]. Cortez [37] analyzed the
phytoplankton community from Bil reservoir in the same
period of this study and it was found a biovolume dominance of Dinophyceae, followed by Cyanophyceae class.
According to Napolitano [52], dinoflagellates can synthesize high amounts of EPA and DHA. Furthermore, most
freshwater zooplankton species have limited or non-ability
to synthesize n-3 PUFA de novo, although they are able to

13

 Lipids
Table2Total fatty acid (FA) profile of stomach content and adipose tissue and triacylglycerol (TAG) of liver and muscle of A. fasciatus (Af)
from the mesotrophic (Ponte Nova, PN) and hypereutrophic reservoirs (Billings, Bil) (MeanSD)
FA (mole %)

Af stomach content

Af adipose tissue

Af hepatic TAG

Af muscle TAG

PN

Bil

PN

Bil

PN

Bil

PN

Bil

n=26

n=34

n=26

n=33

n=31

n=31

n=29

n=29

C15:0
C15:0iso
C15:0anteiso
C16:0iso
C17:0
C17:0anteiso
C18:0iso
C18:0anteiso
C20:0iso
OFA-BFA
C14:0

0.20.10
0.50.28
0.20.04
0.20.14
0.30.22*
0.30.28
0.10.01
0.30.18
0.30.36
2.60.78*
1.01.67

0.50.25
0.50.34
0.40.19
0.20.10
1.90.74
0.60.56
0.20.10
0.50.20
0.40.44
4.51.30
2.91.72

0.20.09
0.40.25
0.20.01
0.30.23
0.30.14*
Nf*
Nf*
Nf*
0.20.05
1.60.40*
1.30.72

0.50.10
0.50.21
0.20.03
0.20.04
1.30.35
0.30.38
0.10.05
0.10.01
0.50.35
3.40.71
2.80.77

0.30.14
0.30.11
0.30.17
0.20.04
0.70.18
0.20.09
0.20.11
0.20.09
0.20.13
2.80.44
1.50.90

0.50.12
0.40.15
0.20.14
0.20.05
1.00.18
0.30.08
0.30.18
0.20.11
0.50.22
4.20.64
2.10.41

0.30.14
0.10.01
Nf
0.20.03
Nf
0.40.11
0.40.14
0.50.20
0.20.04*
1.90.44*
1.10.40

0.40.11
0.40.19
0.30.17
0.20.04
Nf
0.60.34
0.20.14
0.20.08
0.70.35
3.00.85
2.10.54

C16:0
C18:0
C20:0
SFA
C16:1n-7
C18:1n-9
C18:1n-7
C20:1n-9
MUFA
C18:3n-3
C18:4n-3
C20:3n-3
C20:4n-3
C20:5n-3
C22:5n-3
C22:6n-3
PUFA n-3
C18:2n-6
C18:3n-6
C20:2n-6
C20:3n-6
C20:4n-6
C22:2n-6
C22:4n-6
C22:5n-6
PUFA n-6
n3/n6
EPA/ARA

19.85.14*
5.52.10*
0.40.11
26.76.17*
2.11.12
36.010.83*
1.10.32
0.40.17
40.610.86*
1.50.9*
0.30.37*
0.10.01
0.20.15
0.40.36*
0.30.21
0.30.47*
3.01.49*
23.710.93*
0.10.08
0.30.27
0.20.17
0.60.75
Nf
Nf
Nf
25.610.81*
0.10.06*
0.70.47*

27.77.71
8.42.32
0.60.30
39.310.30
7.92.38
18.75.43
5.51.54
0.50.35
32.95.70
9.15.76
1.00.54
0.30.18
0.60.47
4.23.36
0.50.38
1.11.24
14.710.87
4.93.00
0.30.14
0.60.50
0.20.09
1.20.99
Nf
Nf
Nf
7.54.33
1.71.10
3.10.87

24.83.14
6.51.27
0.20.09
32.93.44
3.91.58
39.94.91*
1.71.18
0.70.16
47.22.78*
1.20.91*
0.10.08
0.30.33
0.10.08
0.30.20*
0.20.22
0.40.37*
2.61.83*
14.23.53*
0.30.13
0.30.12
0.40.12
0.50.26
Nf
0.10.10
Nf
16.23.59*
0.20.22*
0.60.23*

26.55.23
7.21.53
0.40.15
36.96.93
8.01.28
24.95.53
4.41.07
0.60.37
38.74.36
6.03.38
0.90.56
0.20.12
0.50.25
2.11.00
0.80.46
1.81.58
13.26.24
5.12.01
0.30.09
0.20.10
0.20.08
1.00.43
Nf
0.20.10
0.20.13
7.32.66
1.80.63
2.30.73

25.24.34
5.52.20
0.20.07
32.84.73
5.11.65
42.06.14*
1.80.50
0.60.21
49.55.87*
0.70.39*
0.30.25
0.10.04
0.30.21
0.40.24*
0.20.12
0.80.57*
2.81.42*
8.73.72*
0.70.56
0.50.21
0.50.29
0.70.34
0.30.20
0.10.04
0.20.17
11.82.39*
0.20.18*
0.60.38*

25.72.6
6.61.55
0.30.12
34.73.42
8.41.41
22.44.54
4.71.13
0.40.22
35.84.22
6.82.12
0.70.35
0.40.18
0.60.28
2.20.75
1.00.59
3.12.73
15.03.88
4.91.13
0.90.66
0.50.37
0.40.25
1.40.64
0.30.28
0.20.12
0.20.19
8.01.55
1.90.45
1.80.77

22.62.28
7.10.70
0.20.11
31.02.01
3.80.78
36.15.95*
2.00.53
0.60.14
42.45.47*
1.20.54*
0.40.20
0.30.09
0.70.54
1.11.23*
0.40.28
2.61.90
6.74.20*
11.72.10*
0.50.29
0.70.36
0.60.17
1.70.89
0.50.54
0.30.25
0.50.50
16.42.02*
0.40.23*
0.60.29*

24.22.44
7.51.00
0.40.10
34.13.09
7.40.99
21.33.46
4.80.89
0.40.16
33.93.31
8.31.98
0.70.25
0.30.09
0.70.47
3.10.94
0.90.39
3.72.67
18.43.57
5.30.89
0.50.19
0.60.50
0.30.15
1.40.51
0.40.43
0.20.15
0.30.28
9.01.65
2.10.35
2.40.71

PUFA total

29.811.51

23.414.13

19.33.29

21.58.49

15.34.87*

25.44.80

24.35.48

28.64.54

n total number of individuals analyzed/species/reservoir, Nf not found, OFA odd chain fatty acid, BFA branched chain fatty acids, SFA saturated
fatty acid, MUFA monounsaturated fatty acid, PUFA polyunsaturated fatty acid, EPA eicosapentaenoic acid, ARA arachidonic acid
*Significant differences at 95% confidence limit (Student t test)

13

Lipids
Table3Fatty acid (FA) profile of phospholipds (PL) of liver and muscle of A. fasciatus (Af) and H. malabaricus (Hm) from the mesotrophic
(Ponte Nova, PN) and hypereutrophic reservoirs (Billings, Bil) (MeanSD)
FA (mole %)

Af hepatic PL

Af muscle PL

Hm hepatic PL

Hm muscle PL

PN

Bil

PN

Bil

PN

Bil

PN

Bil

n=20

n=31

n=21

n=31

n=12

n=21

n=19

n=24

C15:0
C15:0iso
C15:0anteiso
C16:0iso
C17:0anteiso
C18:0iso
C18:0anteiso
C20:0iso
OFA-BFA
C14:0
C16:0

0.30.11
0.30.06
1.00.84
0.40.11
0.20.07
0.20.08
0.30.12
0.40.25
3.00.41
0.50.26
14.84.25

0.30.10
0.20.06
0.20.09
0.50.15
Nf
0.20.04
0.30.09
0.20.15
2.00.38
0.60.37
17.15.34

0.20.18
Nf
Nf
0.90.28
Nf
1.30.65
1.70.53
0.20.19
4.30.91
0.40.22
19.24.34

0.20.13
Nf
Nf
1.10.36
Nf
1.10.70
1.60.60
0.30.14
4.40.92
0.50.23
18.34.93

0.60.35
0.30.14
Nf
0.50.18
0.50.27
0.50.09
0.50.34
Nf
2.01.00
0.50.29
17.36.71

0.40.19
0.40.25
Nf
0.20.10
0.20.06
0.40.17
0.20.16
0.20.08
2.20.55
0.90.68
19.45.94

0.40.15
Nf
Nf*
2.30.93
0.40.09*
0.80.50
1.20.35
0.40.22
5.01.51*
0.50.20
14.73.45

0.20.11
0.20.01
2.60.05
2.10.64
2.83.57
1.00.38
1.30.24
0.30.09
9.22.25
0.40.14
13.42.79

C18:0
C20:0
SFA
C16:1n-7
C18:1n-9
C18:1n-7
C20:1n-9
MUFA
C18:3n-3
C18:4n-3
C20:3n-3
C20:4n-3
C20:5n-3
C22:5n-3
C22:6n-3
PUFA n-3
C18 n-3
C20-22 n-3
C18/20-22 n-3
C18:2n-6
C18:3n-6
C20:2n-6
C20:3n-6
C20:4n-6
C22:2n-6
C22:4n-6
C22:5n-6
PUFA n-6
C18 n-6
C20-22 n-6
C18/20-22 n-6
n3/n6

18.92.82
0.20.10
34.54.5
1.70.59
16.53.79*
1.70.40
0.90.57
20.94.62
0.50.21*
0.30.22
0.30.32
0.40.41
1.30.78*
0.80.35
12.25.7
16.86.63*
0.70.29*
16.06.48*
22.010.42
5.01.60*
1.71.02
1.90.54
2.50.80
11.73.53*
0.50.30
1.70.91
0.40.38
25.54.33*
6.71.92*
18.84.23*
3.31.36
0.70.31*

18.43.10
0.30.14
36.56.91
3.01.58
12.95.25
4.01.19
0.40.16
20.27.11
2.81.20
0.40.24
0.70.32
0.60.23
5.02.40
2.51.28
15.37.72
29.110.88
3.01.25
25.18.08
105.22
2.51.12
1.71.62
0.50.23
0.70.30
6.72.98
0.10.21
0.60.6
0.10.22
13.03.57
4.11.64
9.33.44
2.61.42
2.20.69

16.34.00
0.10.13
36.15.63
1.10.52
9.53.16
2.21.94
0.30.20
12.92.38
0.50.27*
0.20.3
0.10.11
0.20.46
2.81.00*
1.70.51
18.55.31
25.16.33*
0.70.36*
24.56.21*
44.519.36
5.41.77*
0.80.32
0.80.50
1.50.75
10.61.67*
0.10.13
2.20.84
0.10.23
22.74.16*
6.21.74*
15.62.96*
2.90.89
1.30.42*

16.51.71
0.20.14
35.04.91
1.50.58
7.42.11
3.01.15
0.20.20
12.62.38
1.90.86
0.20.22
0.30.16
0.70.23
6.71.77
3.00.84
23.96.25
37.06.77
2.01.25
36.06.14
17.68.31
2.11.29
0.90.46
0.40.28
0.60.20
6.20.81
0.10.18
1.00.40
0.10.18
11.31.01
3.00.86
8.91.15
2.80.94
3.20.53

19.73.80
0.30.27
37.07.31
2.41.31
8.44.25
5.72.98
0.30.18
16.85.39
0.80.23
0.71.31
0.40.14
0.30.17
2.71.54
1.60.57
14.16.30
22.17.99
1.41.38
20.77.82
2720.37
3.81.64
1.20.64
0.50.36
0.90.31
12.84.73
0.20.67
1.91.68
0.10.34
22.35.94
4.81.95
17.55.74
4.22.05
1.00.31

21.85.97
0.40.22
41.97.26
3.01.88
9.06.03
6.82.93
0.50.37
19.07.17
1.70.68
0.20.14
0.60.21
0.40.19
4.21.87
1.71.12
12.27.06
21.19.27
1.80.75
21.49.37
14.17.41
2.20.81
1.40.64
0.40.26
0.50.14
8.23.73
0.20.51
1.20.81
0.10.27
14.94.48
3.51.15
11.44.38
3.72.23
1.50.42

18.14.04
0.10.15
32.56.25
1.60.48
7.31.71*
2.60.55
0.30.24
11.52.34
0.60.31
0.40.29
0.20.16
0.10.23
1.70.69
2.81.02
23.05.04
30.86.11
0.90.34
31.86.12
39.415.95
2.90.85
1.00.33
0.40.33
0.70.28
11.42.18*
0.10.22
2.41.62
0.51.33
20.33.56*
3.80.94
16.63.21*
4.61.37
1.70.45*

17.52.11
0.20.16
31.83.11
1.50.59
4.41.62
5.42.66
0.40.26
11.82.16
1.00.31
0.82.05
0.40.13
0.60.97
2.60.58
3.31.29
23.74.41
35.43.40
1.81.88
32.64.15
28.813
1.40.78
1.00.30
0.30.18
0.40.13
7.41.34
0.10.11
2.40.86
0.10.15
14.01.69
3.21.54
11.81.72
4.31.26
2.50.34

13

 Lipids
Table3continued
FA (mole %)

EPA/ARA
PUFA total

Af hepatic PL

Af muscle PL

Hm hepatic PL

Hm muscle PL

PN

Bil

PN

Bil

PN

Bil

PN

Bil

n=20

n=31

n=21

n=31

n=12

n=21

n=19

n=24

0.80.22

0.30.11*

1.10.31

0.20.11

0.50.14

0.20.08*

0.40.09

41.212.13

47.55.10

48.17.08

43.012.51

36.812.54

50.87.26

48.04.05

0.10.06*
42.67.26

n total number of individuals analyzed/species/reservoir, Nf not found, OFA odd chain fatty acid, BFA branched chain fatty acids, SFA saturated
fatty acid, MUFA monounsaturated fatty acid, PUFA polyunsaturated fatty acid, EPA eicosapentaenoic acid, ARA arachidonic acid
*Significant differences at 95% confidence limit (Student t test)

accumulate, select and biotransform dietary FA [2527, 31]

especially the microzooplankton (ciliates and heterotrophic
flagellates), contributing also to the n-3 LC-PUFA transfer
to higher trophic levels [53].
The presence of zooplankton and aquatic insects in the
stomach content of A. fasciatus from the hypereutrophic
reservoir (Bildata not shown) could suggest trophic
transfer of n-3 LC-PUFA from zooplankton, probably via
dinoflagellates, to the fish captured, and a possible biotransformation of 18:3n-3 to EPA and DHA by the zooplankton diet, prior to the transference. Guarapiranga reservoir, a hypereutrophic reservoir located close to Billings,
presented a zooplankton biomass 25-fold greater and with
dominance in biomass of cyclopoids compared to the Ponte
Nova reservoir during the dry season, which displayed a
dominance of cladocerans throughout the year [54]. In both
situations, zooplankton components seem to be an important link to higher trophic levels in the hypereutrophic Bil
reservoir. Interestingly, 18:2n-6, the main n6-PUFA found
in the fish sampled from the mesotrophic reservoir (PN), is
also usually the main PUFA in higher plants [55]. Hence,
the higher concentration of this FA in the stomach content
of A. fasciatus suggests a diet rich in vegetal sources, a fact
corroborated by the predominance of higher plant material
(leaves and seeds) found in the stomach contents from fish
caught from the PN reservoir (data not shown). Since n-3
and n-6 PUFA have been considered as indicators of the
autochthonous (e.g. phytoplankton) and the allochthonous
origin of food resources [29, 52, 56], the observed differences in FA profiles between A. fasciatus diets from both
reservoirs may reflect changes in the origin of the main dietary source, from mainly allochthonous in the mesotrophic
reservoir, to mainly autochthonous in the hypereutrophic
reservoir.
Differences observed for PUFA profiles of the stomach content from fish in both reservoirs were basically the
same, regardless of the fish species. In other words, it is
always possible to observe higher n-6 than n-3 PUFA contents in fish from the PN reservoir (mesotrophic) and higher
n-3 than n-6 PUFA contents in fish from the Bil reservoir
(hypereutrophic). This result corroborates the hypothesis

13

that FA profiles of predators can indeed reflect the profile

of their prey, which is in turn affected by environmental
conditions, a classic bottom-up effect [49]. Additionally,
FA profiles found in the stomach content from fish of both
reservoirs seemed to be similarly assimilated and incorporated in all tissues analyzed, with minor alterations related
mainly to the lipid fraction analyzed (TAG or PL) or the
feeding habit. Compared to the FA composition of PL,
TAG stored in animal tissues has a FA composition very
similar to that found in the diet [5759]. The SDA analysis
run for each fish species separately confirms this assumption, as better separations obtained for TAG than PL fatty
acids. However, these results also suggest that this trend
may differ between omnivorous and carnivorous fish species. The high percentages of n-6 C18-PUFA and n-3
PUFA (18 and 2022) in the stomach contents of A. fasciatus from PN and Bil reservoirs, respectively, probably
resulted in the same FA composition found in the different
tissue TAG (adipose tissue, liver, and muscle), while differences found for PL fatty acids occurred mainly due to
higher values of LC-PUFA. In general, the FA composition
of PL seems to be more conservative and less dependent
on prey FA composition, especially in omnivorous species. This could be associated with the physiological role
of PL in cellular membrane, in comparison with the energy
reserve function of TAG in animal tissues [11]. In H. malabaricus, since both stomach content and tissue TAG contained high values of 18 and LC-PUFA (higher n-6 and
higher n-3 FA in PN and Bil reservoirs, respectively), this
could suggest a direct intake of these FA from their prey.
However, no differences were observed in the PL samples
between reservoirs, except those found for ARA percentages. High percentages of ARA, as also of EPA and DHA,
from the diet are expected to accumulate in the TAG of
carnivorous animals, since in the digestion process by
predator, TAG and PL are hydrolyzed in free fatty acids
and monoacylglycerol (and/or diacylglycerol) or lysophospholipids, respectively, and stocked mainly in the adipose
tissue as TAG [49]. As a result, the activity of desaturases
and elongases, even being high in freshwater fish, is usually lower in carnivorous animals than in omnivorous [3].

Lipids
Table4Total fatty acid (FA) profile of stomach content and adipose tissue and triacylglycerol (TAG) of liver and muscle of H. malabaricus
(Hm) from the mesotrophic (Ponte Nova, PN) and hypereutrophic reservoirs (Billings, Bil) (MeanSD)
FA (mole %)

Hm stomach content

Hm adipose tissue

Hm hepatic TAG

Hm muscle TAG

PN

Bil

PN

Bil

PN

Bil

PN

Bil

n=6

n=7

n=5

n=21

n=21

n=28

n=13

n=22

C15:0
C15:0iso
C15:0anteiso
C16:0iso
C17:0
C17:0anteiso
C18:0iso
C18:0anteiso
C19:0
C20:0iso
OFA-BFA

1.60.36
0.40.16
0.30.13
0.40.22
2.00.96
Nf
Nf
0.70.11
Nf
Nf
6.20.86

0.70.31
0.70.37
0.20.05
0.30.14
2.00.54
0.20.07
0.50.39
0.30.12
Nf
0.80.24
6.01.54

1.90.72
0.70.11
0.20.07
0.30.05
1.60.39
Nf
Nf
Nf
Nf
Nf
5.32.18

0.80.19
1.50.50
0.30.09
0.30.09
1.60.24
0.20.06
0.10.04
Nf
Nf
1.00.38
5.31.02

1.20.44
0.80.48
0.40.16
0.40.15
Nf
0.60.5
Nf
0.30.04
1.30.04
0.40.04
5.21.26

0.80.14
1.60.32
0.30.06
0.30.07
Nf
0.30.09
Nf
0.20.12
0.50.24
0.70.31
5.00.70

1.20.51
0.70.25
0.60.30
0.40.21
Nf
0.90.46
0.70.06
0.90.51
0.60.42
1.30.72
6.32.19

0.80.21
1.00.35
0.50.25
0.40.07
Nf
0.90.56
0.60.22
1.10.76
0.50.26
1.80.69
7.02.62

C14:0
C16:0
C18:0
C20:0
SFA
C16:1n-7
C18:1n-9
C18:1n-7
C20:1n-9
MUFA
C18:3n-3
C18:4n-3
C20:3n-3
C20:4n-3
C20:5n-3
C22:5n-3
C22:6n-3
PUFA n-3
C18:2n-6
C18:3n-6
C20:2n-6
C20:3n-6
C20:4n-6
C22:2n-6
C22:4n-6
C22:5n-6
PUFA n-6
n3/n6
EPA/ARA

3.61.87
24.55.88
10.16.48
0.30.10
37.311.26
11.89.78
9.85.19
5.62.42
0.30.16
27.610.11
2.61.21
0.20.22
0.50.30
0.40.27
0.60.24*
1.21.08
5.53.46
12.34.09*
4.83.25
1.10.90
0.70.42
0.80.65
4.61.01
Nf
Nf
Nf
15.18.25
0.70.31*
0.20.12

2.71.05
28.24.18
9.51.92
0.30.10
39.85.47
7.32.51
13.83.84
4.91.48
0.50.15
26.05.96
6.74.12
0.30.31
0.60.30
0.70.33
2.71.71
1.50.74
5.34.20
19.66.48
4.11.68
0.30.19
0.30.12
0.40.23
2.30.91
Nf
Nf
Nf
8.01.91
2.30.70
1.10.47

4.00.63
26.02.39
6.30.92
0.20.04
34.82.74
9.01.14
16.95.82
4.31.00
0.50.12
30.94.48
5.71.85*
0.40.27
0.60.23
0.60.13
1.40.30*
1.60.34
3.91.33*
14.82.39
5.41.26*
1.20.98*
0.50.32
0.80.05
2.80.21
Nf
0.60.17
Nf
13.91.41*
0.90.13*
0.50.22*

3.80.71
23.52.37
6.90.76
0.40.06
33.93.27
9.22.76
16.11.96
4.70.99
0.90.22
31.35.04
8.11.57
0.40.55
0.80.22
1.00.48
2.21.01
2.30.89
6.62.93
20.67.18
4.02.08
0.40.07
0.40.08
0.40.15
1.50.49
0.10.01
0.60.42
0.10.06
7.80.87
2.90.85
1.40.33

3.21.66
24.56.53
10.76.99
0.20.12
38.79.44
8.53.47
15.15.64
4.61.99
0.50.23
30.16.99
1.60.77*
0.60.97
0.30.11
2.02.15
0.70.37*
0.50.19
2.51.57*
9.03.83*
4.82.28*
0.80.64
0.81.05
0.60.33
3.32.87
0.90.90
4.01.78*
0.50.69
16.19.63
0.60.26*
0.30.24*

3.50.83
23.64.44
9.73.68
0.40.22
36.34.55
9.12.93
11.07.07
10.14.66
0.80.38
31.06.01
4.61.58
0.50.33
0.70.26
2.52.57
2.11.03
1.00.56
4.32.97
16.36.35
3.40.73
0.80.43
0.50.21
0.50.18
2.71.89
1.21.32
0.40.28
0.20.17
9.43.13
1.70.57
0.90.44

2.10.80
18.64.24
12.95.70
0.40.17
34.03.14
5.41.69
14.05.43
3.30.72
0.90.49
23.75.88
2.31.22*
1.20.68
0.60.40
1.00.64
0.90.31
1.50.82
7.04.97
15.84.85*
6.01.77*
1.91.17
1.50.89
0.50.23
4.33.14*
1.10.9
1.20.86
0.80.65
19.33.01
0.80.23*
0.30.24*

2.80.72
21.53.51
9.12.40
0.30.14
33.34.01
7.72.78
14.22.59
4.31.69
0.70.26
26.76.42
4.91.33
1.30.84
0.70.28
1.00.76
1.80.75
1.70.89
8.04.63
20.36.52
3.00.85
1.40.86
1.10.68
0.40.20
1.91.09
1.00.86
0.80.51
0.70.69
11.83.46
1.90.62
1.00.37

PUFA total

28.917.28

28.07.68

30.13.46

29.57.53

27.212.61

27.88.44

35.66.15

32.48.32

n total number of individuals analyzed/species/reservoir, Nf not found, OFA odd chain fatty acid, BFA branched chain fatty acids, SFA saturated
fatty acid, MUFA monounsaturated fatty acid, PUFA polyunsaturated fatty acid, EPA eicosapentaenoic acid, ARA arachidonic acid
*Significant differences at 95% confidence limit (Student t test)

13

 Lipids
Table5Standardized canonical coefficients for each fatty acid in
the discriminant functions (canonical roots) extracted by stepwise
discriminant analysis (SDA) carried out to identify fatty acids (independent variables), which significantly separated organisms (groups);
Variables

Eigenvalues (Eigenv.); significance provided by v2-test (p value) and

explained cumulative variance (Cum. r2) are provided for each root
following the fatty acids

TAG A. fasciatus

PL A. fasciatus

TAG H. malabaricus

PL H. malabaricus

Af Hm

Function 1

Function 2

Function 1

Function 2

Function 1

Function 2

Function 1

Function 2

Function 1

Function 2

C16:0
C18:0
C18:1n-9
C18:2n-6
C18:3n-3
ARA
EPA
DHA
Eingenv.

0.07
0.04
0.46
0.76*
0.57*
0.03
0.13
0.13
5.97

0.02
0.66*
0.13
0.60*
0.22
0.55*
0.60*
0.32
0.97

0.07
0.20
0.90*
0.86*
0.20
0.33
0.15
0.19
11.68

0.24
0.42
0.75*
0.81*
0.32
0.67*
0.23
0.65
7.04

0.10
0.20
0.04
0.39
0.70*
0.62*
0.56
0.03
2.47

0.30
0.22
0.27
0.13
0.17
0.81*
0.91*
1.37*
1.13

0.40
0.30
0.16
0.69*
0.28
0.08
0.84*
1.73*
6.27

1.05*
0.02
0.37
0.23
0.18
1.73*
0.20
0.14
2.95

0.05
0.22
0.32
0.58*
0.67*
0.19
0.36
0.03
7.06

0.21
0.24
0.53
0.36
0.36
0.39
0.76*
0.41
2.54

p value

<0.001

<0.001

<0.001

<0.001

<0.001

<0.001

<0.001

<0.001

<0.001

<0.001

Cum. r2

73.2

85.2

54.5

87.3

57.3

83.6

55.9

82.2

61.7

83.9

Fatty acids marked with an asterisk significantly discriminated groups within a function
Af A. fasciatus, ARA arachidonic acid, DHA docosahexaenoic acid, EPA eicosapentaenoic acid, Hm H. malabaricus, PL phospholipids, TAG triacylglycerol

Interestingly, despite the differences found for n-3 PUFA

between reservoirs in this study, DHA percentages were the
same in tissue PL of all samples, suggesting an important
maintenance of appropriate DHA levels for cellular membrane fluidity. Animals from the hypereutrophic reservoir,
which had high proportions of DHA in the TAG, seem to
not transfer this FA in excess to PL. Additionally, despite
lower percentages of n3-PUFA in the stomach content and
storage tissues of animals from the mesotrophic reservoir,
their PL samples had always high percentages of DHA. It
seems that, regardless of dietary DHA availability, physiological necessity to maintain appropriate levels of DHA
in membrane PL helps to diminish differences between PL
samples from both reservoirs.
Both investigated fish species from the hypereutrophic
reservoir had higher n3/n6 and EPA/ARA ratios in all analyzed tissues, compared to the same species from the mesotrophic system. The optimal physiological n3/n6 ratio is not
known for most organisms, but it has been hypothesized to
be species-specific [60]. Changes in dietary n3/n6 ratios
can directly modify EPA/ARA ratios in fish tissues [61],
and our results corroborate this statement for both investigated fish species. ARA and EPA act as precursors in the
eicosanoid synthesis, playing the role of local hormones
or signaling molecules in the control of inflammation and
immunity responses [13, 62, 63], hematological and cardiovascular activity, renal and neural function [11], as well as
reproduction [21, 64]. However, ARA-derived eicosanoids
are more physiologically active compared to EPA-derived
eicosanoids, so that modifications in cell EPA/ARA ratios

13

can result in differentiated physiological responses of the

two eicosanoid series [11, 20, 61, 65]. Once dietary PUFA
composition had only minor effects in H. malabaricus
EPA/ARA ratios (which were only 0.5 times higher in fish
from the hypereutrophic system, than in those from the
mesotrophic system), such consequences for physiological
responses may be less expected in this carnivorous species
(EPA/ARA ratios were three times higher in A. fasciatus
from the hypereutrophic system, compared to the same
species from the mesotrophic system). Studies comparing
n3/n6 and DHA/ARA ratios in an herbivorous-omnivorous
(Rutilus rutilus) and a carnivorous fish (Perca fluviatilis)
from oligotrophic and eutrophic lakes found significantly
higher n3/n6 in the carnivorous animals from the eutrophic
lake [66]. The authors concluded that omnivorous fish were
more flexible in relation to dietary items than carnivorous
fish, which feed always on similar items and, therefore, on
similar FA composition.
These data allow us to assume that the nutritional and/
or physiological demand of consumers selectively and efficiently changes according to habitat and dietary conditions,
but this ability depends on phylogenetic origin and/or life
history [25]. Recent studies from temperate lakes showed
a decrease in saturated FA and 18-PUFA and an increase of
n-3 LC-PUFA from lower trophic levels towards predators
[29, 31]. We also found an increase of LC and a decrease
in 18-PUFA in the investigated fish species from both reservoirs, compared to the FA composition of stomach contents. Thus, the results of the present study suggest a differential deposition of FA in the animal tissues resulting

Lipids

Fig.2Canonical functions provided by stepwise discriminant analyses (SDA) run separately for A. fasciatus using a triacylglycerol
(TAG) and b phospholipids (PL). See text and Table5 for linear combinations of fatty acid discriminating groups within each function. AT
adipose tissue, HPL hepatic phospholipids, HTAG hepatic triacylglycerol, MPL muscle phospholipids, MTAG muscle triacylglycerol, SC
stomach contents

from dietary FA modification, regardless of environmental

trophic state.
It is known that phytoplankton community composition can determine PUFA available to consumers, but not
all PUFA are transferred equally throughout the food chain.
The trophic transfer of PUFA in pelagic environment from
large lakes may be highly selective, and patterns of transfer, mainly of n-3 PUFA, depend on FA molecular structure [31]. Thus, as the hypothesis predicted, changes in
planktonic community composition due to eutrophication
are expected to affect PUFA transfer patterns in the food
web, mainly due to reduced n-3 PUFA availability. However, the results of the present study refuted this hypothesis, since no n-3 PUFAs deficiency (mainly of EPA and

Fig.3Canonical functions provided by stepwise discriminant analyses (SDA) run separately for H. malabaricus using a triacylglycerol
(TAG) and b phospholipids (PL). See text and Table5 for linear combinations of fatty acid discriminating groups within each function. AT
adipose tissue, HPL hepatic phospholipids, HTAG hepatic triacylglycerol, MPL muscle phospholipids, MTAG muscle triacylglycerol, SC
stomach contents

DHA) was observed in the dietary items from the hypereutrophic reservoir (inferred by the fish stomach contents).
Moreover, the differences observed in the FA profile of the
investigated fish species were more dependent on endogenous factors, such as (1) feeding habit (carnivorous species accumulated more LC-PUFA in their tissues, while
omnivorous species presented higher accumulation of
18-PUFA), and (2) fatty acids selective metabolism, involving accumulation and biotransformation of FA in specific
tissues and lipid classes, as observed for DHA and ARA in
the tissues PL. Trophic transfer of n-3 18 and LC-PUFA in
pelagic food webs may depend upon taxon-specific feeding strategies (e.g., filter feeders vs. selective feeders)
and/or selective metabolism of FA in consumer, including

13

 Lipids

References

Fig. 4a Canonical functions provided by stepwise discriminant analyses (SDA) run separately for A. fasciatus H. malabaricus using
triacylglycerol (TAG). See text and Table5 for linear combinations of
fatty acid discriminating groups within each function. AT adipose tissue, HTAG hepatic triacylglycerol, MTAG muscle triacylglycerol, SC
stomach content. b PUFA composition divided into groups LC-PUFA
and 18-PUFA of A. fasciatus H. malabaricus TAG from the both
reservoir. abSignificant differences at 95% confidence limit (Student
t test)

selective retention, biosynthesis, catabolism and modification of FA [31]. Futures studies are clearly necessary to
enhance the understanding about this intricate and dynamic
trophic transfer of n-3 PUFA in tropical reservoirs, especially under the threat of increasing eutrophication and land
use changes.
Acknowledgments The present study was funded by the Fundao
de Incentivo Pesquisa do Estado de So Paulo (FAPESP) (Research
Grant: 2012/50371-2; Ph.D. scholarship: 2010/50555-0) and by
the Comisso de Aperfeioamento de Pessoal do Nvel Superior
(CAPES). The authors would like to thank the fishermen Evaldo
Bizarrias and Milton Nunes de Santana and the LAMEROA team, for
their help in fish collection, and IB/USP for providing logistics and
facilities for collection.

13

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