JOURNAL OF THE Vol. 37, No.

3
WORLD AQUACULTURE SOCIETY September, 2006

Effect of Diets Formulated with Native Peruvian Plants

on Growth and Feeding Efficiency of Red Pacu
(Piaractus brachypomus) Juveniles
MARIA E. PALACIOS,1 KONRAD DABROWSKI,2 MARY ANN G. ABIADO,3
AND KYEONG-JUN LEE4
The Ohio State University, School of Environment and Natural Resources, 2021 Coffey Road,
Columbus, Ohio 43210 USA

CHRISTOPHER C. KOHLER
Southern Illinois University, Fisheries and Illinois Aquaculture Center, Carbondale, Illinois
62901 USA

Abstract
We evaluated the effects of casein-based semipurified diets, alone or supplemented with native
Peruvian plants, on growth, feed efficiency, and histology of the digestive tract of red pacu, Piaractus
brachypomus, juveniles over an 8-wk feeding trial. Three tanks were randomly assigned to one of four
caseingelatin (40:8) diets containing a supplement of 15% wheat meal (control) or an identical level of
substitution of three South American native plant as follows: camu-camu fruit (Myrciaria dubia),
aguaje fruit (Mauritia flexuosa), or maca tuber meal (Lepidium meyenii). The fish (initial weight,
2.04 6 0.06 g) were fed experimental diets at decreasing feeding rates from 4 to 2.6% of body weight.
After 8 weeks of feeding, fish fed a diet supplemented with maca meal showed significantly higher
(P , 0.05) weight gain, specific growth rate, protein efficiency ratio (PER), apparent net protein
utilization (NPU), and instantaneous feed intake than fish fed other diets. Feed conversion ratio (FCR),
PER, and NPU in fish fed the caseingelatin diet supplemented with maca meal were among the best
ever reported in the scientific literature, 0.64 6 0.03, 3.13 6 0.15 and 23.8 6 2.0, respectively. The
camu-camu meal had a negative impact on diet palatability and utilization, which resulted in slower
growth. The stomach, intestine, pancreas, and pyloric caeca at the start and end of the experiment
showed normal differentiation and appearance of cells and tissues. The liver parenchyma showed
lipid infiltration and pigment accumulation in all samples at the initiation of the experiment and may
be attributed to the period of decreased feed intake prior to the study. At the end of the study,
similar histopathologies were recorded in all samples from the control and camu-camu groups.
Normal liver histology (polyhedral hepatocytes with centrally located nuclei) was observed in
two of three samples from the maca group and all the samples from the group that was fed the
aguaje-supplemented diet.

Plants from the Peruvian Andes, such as
maca, Lepidium meyenii (Brassicaceae), and
the Amazonian rainforest fruit species, such as
aguaje, Mauritia flexuosa (Arecaceae), and
camu-camu, Myrciaria dubia (Myrtaceae), are
widely used for human consumption and for
1 Present address: Universidad Nacional Mayor de San
Marcos, CP 14-02 Lima 14, Peru.
2 Corresponding author.
3 Present address: Department of Plant Cellular and
Molecular Biology, 318 W. 12th Avenue, Columbus, Ohio
43210 USA.
4 Present address: Faculty of Applied Marine Science, Cheju
National University, Jeju-Do, Jeju, South Korea 690-756.
Copyright by the World Aquaculture Society 2006
medicinal purposes because of their high-
nutritional quality and antioxidant properties
(Leon 1964; Dini et al. 1994; Moraes et al.
1994; Obregon 1998). Dispersal fruits and seeds
of tropical plant species have been reported
as part of the diet of frugivorous fish of the
Amazon (Araujo-Lima and Goulding 1998;
Roubach and Saint-Paul 1994).
Maca has been consumed since the Pre-
Hispanic period by humans inhabiting elevated
altitudes in the highlands of Peru (Leon 1964).
The plant contains several phytochemicals such
as phytosterols, aromatic glucosinolates, sapo-
nins, and flavonoids, which can potentially

246
 EFFECTS OF CASEIN-BASED DIETS ON GROWTH, FEED EFFICIENCY OF RED PACU
increase metabolic rate, improve fertility, and
serve as antioxidants (Cicero et al. 2001; Gonzales
et al. 2001; Li et al. 2001; Sandoval et al. 2002).
Camu-camu is a globose berry fruit from the
Western Amazon rainforest that contains the
highest concentration of vitamin C (36 g of
ascorbic acid per 100 g dry matter) known
in the plant kingdom (Moraes et al. 1994).
Camu-camu has astringent, antioxidant, anti-
inflammatory, and nutritive properties. The
palm of aguaje is a massive palm tree (100 feet)
from the Amazon rainforest, and the fruits
are eaten raw or processed and added to
beverages and ice creams. It is also consumed
by wild primates, rodents, and other mammals
in the rainforest (Padoch 1988; Penn and Neise
2005).
The red belly pacu, Piaractus brachypomus,
originates from the Amazon basin of South
America. This fish has excellent potential for
aquaculture because of its high growth rates in
earthen ponds or floating cages (Melard et al.
1993). It is an important ornamental species in
Central and South America and, as such, com-
mands a high market price in Western countries
(Baras 2000; Sakamoto et al. 2001). However,
uncontrolled commercial exploitation and ille-
gal poaching may cause significant reduction
in founder populations and potential inbreeding.
Therefore, efforts to study various aspects of the
culture of red pacu need to become a priority.
Currently, there is only limited information
available on the nutritional requirements and
the suitability of locally available plants as die-
tary ingredients for fishes of the genus Piaractus
(Fernandes et al. 2001; Fracalossi 2002; Vasquez-
Torres et al. 2002). Histological analysis of
the digestive tract of pacu is important in devel-
oping appropriate formulated diets for this
species and to determine whether the experi-
mental diets can lead to pathologies of the diges-
tive tract that can compromise the digestion and
absorption of nutrients.
This study aims to evaluate the effects of
dietary supplementation of semipurified casein
gelatin-based diets with maca, aguaje, and camu-
camu meal on growth, feed, and protein
utilization and to determine the histological
changes of the digestive tract of red pacu juveniles.
247
Materials and Methods
Experimental Diets
Semipurified, isonitrogenous caseingelatin
(40:8%)-based diets were supplemented with
15% wheat meal (control) or with one of three
other Peruvian native plants as follows: camu-
camu meal, aguaje meal, or maca meal
(Table 1). Camu-camu fruits were collected
fresh in the vicinity of fishponds (Instituto de
Investigaciones dela Amazonia Peruana,
Pucallpa, Peru), frozen, and then freeze-dried
to be used on a dry basis. Aguaje pulp (col-
lected as yellow cake for human consumption;
mesocarp; see Padoch 1988) was purchased at
the local market in Iquitos (Peru) and kept fro-
zen until freeze-dried and used as dietary
ingredient. Maca meal was processed in
the Agriculture University La Molina, Lima,
Peru, and supplied as dry powder. The diets
were also supplemented with 5% fish protein
concentrate (CPSP 90, Sopropeche S.A., Bou-
logne-Sur-Mer, France) to enhance palatability
of the semipurified experimental diets (Dab-
rowski et al. 2003). The diets were cold-pel-
leted (2.0-mm diameter) and freeze-dried to
have less than 5% moisture. Pellets were
ground and sieved to obtain particle sizes of
0.50.7 and 0.71 mm for use during the first
2 wk and until the end of the experimental
period, respectively.
Fish and Experimental System
Red pacu juveniles (2.04 6 0.06 g mean ini-
tial weight) were purchased from Ekk Will Water
Life Resources (Gibsonton, FL, USA). Fish (20
per tank) were distributed in twelve rectangular
glass aquaria supplied with UV-irradiated and
filtered, semirecirculated, dechlorinated city water
at a flow rate of 1.01.5 L/aquarium/min. Three
aquaria were randomly assigned to one of four
diets, and the feeding trial was conducted for
8 wk. Fish were fed experimental diets at a feed-
ing rate gradually decreasing from 42.6% of
body weight as follows: 4% at the beginning,
3.5% at the fourth week, 2.8% at the sixth week,
and 2.6% until the end of the feeding period. The
fish were fed three times per day at 10:00, 13:30,
and 17:00 h, 7 d/wk. Feeding rate and frequency
248 PALACIOS ET AL.

TABLE 1. Composition and proximate analyses of the experimental diets used in the experiment (% dry matter).

Diets
Ingredients Control (D1) Camu-camu (D2) Aguaje (D3) Maca (D4)
Casein1 40.00 40.00 40.00 40.00
Gelatin1 8.00 8.00 8.00 8.00
Dextrin1 15.00 15.00 15.00 15.00
Wheatmeal2 15.00
Camu-camu meal3 15.00
Aguaje meal4 15.00
Maca meal5 15.00
Soybean oil1 5.00 5.00 5.00 5.00
Cod liver oil1 5.00 5.00 5.00 5.00
Vitamin mix6 4.00 4.00 4.00 4.00
Mineral mix7 3.00 3.00 3.00 3.00
Ascorbic acid8 0.02 0.02 0.02 0.02
Carboxymethylcellulose1 2.00 2.00 2.00 2.00
L-Arginine1 0.50 0.50 0.50 0.50
L-Methionine1 0.40 0.40 0.40 0.40
L-Lysine1 0.80 0.80 0.80 0.80
Choline chloride1 1.00 1.00 1.00 1.00
Fish protein concentrate9 5.00 5.00 5.00 5.00
Crude protein 50.67a 50.74a 49.41a 50.06a
Ash 4.54b 4.42bc 4.36c 4.82a
Dry matter 97.48a 95.97b 96.97a 95.12b
Mineral composition
Calcium (%) 0.55b 0.52c 0.56b 0.61a
Phosphorus (%) 0.75a 0.73b 0.72b 0.77a
Potassium (%) 0.39d 0.48c 0.51b 0.67a
Sodium (%) 0.37b 0.45a 0.28c 0.27d
Copper (mg/g) 4.50b 7.70a 4.35b 4.30b
Iron (mg/g) 80.80ab 92.30a 53.40b 53.40b
Magnesium (mg/g) 175.00b 221.50b 268.00a 268.50a
Manganese (mg/g) 214.75a 103.40b 110.45c 101.45c
Zinc (mg/g) 28.05b 28.65b 28.70b 32.40a
Means of triplicate groups; values (6SD) with different superscript letters in the same row are significantly different
(P , 0.05).
1 ICN Biomedicals (Costa Mesa, CA, USA).
2 Wheat meal chemical composition (%): crude protein, 11.7; crude lipid, 1.2; and ash, 0.4.
3 Camu-camu meal chemical composition (% dry matter basis): crude protein, 8.0; crude lipid, 1.5; and ash, 5.3.
4 Aguaje meal chemical composition (% wet basis): crude protein, 4.0; crude lipid, 8; and ash, 0.9.
5 Maca meal chemical composition (% wet basis): crude protein, 13.7; crude lipid, 5.22; ash, 5.57; moisture, 3.75; P,

0.34; K, 1.79; Ca, 0.44; Mg, 0,087; Fe, 0.0086; Na, 0.0235; Zn, 0.0043; and Mn, 0.0017.
6 Roche performance premix composition per g of vitamin mixture: vitamin A, 2646 IU; vitamin D, 221 IU; vitamin E,

66.1 IU; vitamin B, 13 mg; riboflavin, 13.2 mg; niacin, 61.7 mg; D-pantothenic acid, 22.1 mg; menadione, 1.32 mg; folic
acid 1.76 mg; pyridoxine, 4.42 mg; thiamine 7.95 mg; D-biotin, 0.31 mg (Hoffman-La Roche, Nutley, NJ, USA).
7 Five milligrams Se in the form of sodium selenite per kg Berhardt Tomarelli salt mixture (ICN).
8 Mg-L-ascorbyl-2-phosphate (Showa Denko K.K., Tokyo, Japan).
9 CPSP 90 (Sopropeche, S.A., Boulogne-Sur-Mer, France).

were based on previous results tested with
tambaqui (Colossoma macropomum), which is
systematically related to Piaractus (Van der
Meer et al. 1997). Temperature was maintained
at 27.5 6 0.1 C. Feces were siphoned daily, and
thorough cleaning of tanks was done weekly.
Diurnal light:dark cycle was regulated at
13:11 h. Feeding was stopped 24 h prior to
weighing. Feeding experiments were conducted
in the Aquaculture Laboratory, School of Envi-
ronment and Natural Resources, The Ohio State
University, Columbus, Ohio, USA, following
animal protocols approved by the Institutional
Animal Care and Use Committee.
 EFFECTS OF CASEIN-BASED DIETS ON GROWTH, FEED EFFICIENCY OF RED PACU
Sample Collection and Analyses
At the beginning of the experiment, sixteen
fish were randomly sampled to determine initial
weight, whole-body proximate composition,
and mineral composition.
Significant differences in diet acceptance
were observed after the first few days of
feeding. The palatability of the camu-camu
diet was compromised; hence, the diet was
neutralized to pH 7.0 using sodium hydroxide
and reprocessed. Previous studies showed that
adjustment of pH in formulated diets did not
compromise fish performance (Whiteman
and Gatlin 2005). Amounts of food were read-
justed in the case of this treatment, and no feed
waste was observed. Every 2 wk, weight gain
was evaluated in each tank and the amount
of food was readjusted. Every 4 wk, one fish
per aquarium and three fish per dietary treat-
ment were euthanized in 100 mg/L tricaine
methane sulfonate (MS-222) for histological
examinations. At the end of the feeding trial,
all fish were weighed and percent weight gain
(body weight gain 3 100/initial body weight),
feed conversion ratio (FCR; dry feed con-
sumed/body weight gain), protein efficiency ratio
(PER; body weight gain/protein intake), specific
growth rate ([ln final weight ln initial weight]
3 100]/d), and mortalities were calculated.
A feed acceptance test (instantaneous feed
intake) was conducted at Days 28 and 42 and
at the end of the feeding trial to determine the
palatability effect of different plant supplements
on diet composition. Fish were fed ad libitum,
and the amounts of feed that fish consumed were
calculated as a percentage of fish weight, and the
mean (6SD) was calculated. After overnight
fasting, the test was conducted by feeding the
fish to apparent satiation during the morning
of Days 28 and 42, and for 4 consecutive d at ter-
mination of the study. A different observer car-
ried out each test.
Three fish carcasses per tank as one statistical
unit, or a total of nine fish per dietary treatment,
were used for whole-body proximate and min-
eral analyses. Analyses of proximate composi-
tion were performed by standard procedures
(AOAC 1995). Protein was calculated as N 3
249
6.25, and apparent net protein utilization
(NPU) was calculated as NPU 5 body protein
gain (g) 3 100/protein intake (g). Mineral com-
position of diets and whole body were deter-
mined by the inductively coupled plasma
emission spectrophotometric method using an
ARI-3560 Spectrometer (Applied Research
Labs, Valencia, CA, USA) according to Watson
and Isaac (1990).
Histological Analyses
One fish per aquarium (three per treatment)
were collected during the initial and final sam-
pling. Fish were euthanized in 100 ppm tricaine
methane sulfonate (MS-222) and fixed in 10%
phosphate-buffered formalin for histological anal-
ysis. After 2 d, the head and the tail regions from
each fish were removed, and the body was trans-
ferred to 70% ethyl alcohol and stored at room
temperature until further processing. Samples
were dehydrated in alcohol, embedded in paraffin,
sectioned (10 mm), and stained in hematoxylin
eosin (Humason 1979). The slides were examined
using the Zeiss Axioskope (Carl Zeiss, Inc.,
Thornwood, NY, USA) equipped with a digital
camera. The images were captured and saved
using the Axiovision digital imaging software
(Carl Zeiss). The appearance of the digestive tract
(liver, stomach, intestine, and pancreas) among
the samples from each treatment group was char-
acterized, and significant pathologies were identi-
fied. Histological observations were based on
descriptions given by Groman (1982), Ferguson
(1989), and Roberts (1989).
Data Analyses
Significant differences among dietary treat-
ments were determined using one-way analysis
of variance followed by least significant differ-
ence test of treatment means (P 5 0.05; Steel
and Torrie 1980). Analyses were done using
the SPSS statistical package (Version 12.0,
SPSS, Inc., Chicago, IL, USA).
Results
There was only minor mortality (05% per
tank), and it was not associated with dietary
treatment. Growth differences were significant
250 PALACIOS ET AL.

FIGURE 1. Mean body weight of red pacu juveniles fed four experimental diets supplemented with 15% of wheat meal
(control), 15% of wheat camu-camu meal, 15% of aguaje meal, and 15% of maca meal for 8 wk. Values are means 6 SD
(n 5 3). Means annotated with different letters are significantly different (P , 0.05).

between treatment groups by 2 wk into the trial
(Fig. 1). After 8 wk, significant differences
(P , 0.05) in individual body weight, weight
gain, and FCR existed (Table 2). Fish fed maca
diet exhibited the highest mean final weight and
weight gain, PER, and NPU whereas fish fed the
camu-camu diet had the lowest growth rate,
PER, and NPU. Fish fed maca diet also showed
the greatest instantaneous food intake (P , 0.05)
following intermittent and final measurements
(Fig. 2). Differences were significant between
the fish fed the maca diet and the remaining
groups (Table 2). Significantly, higher levels
of protein, ash, and minerals were observed in
fish fed the camu-camu diet relative to fish
fed the control, aguaje, and maca diets. The

TABLE 2. Growth and feed utilization parameters of red pacu juveniles fed experimental diets1

Diets
Variable Control Camu-camu Aguaje Maca
Initial body weight (g) 2.06 0.05a 2.06 0.05a 2.06 0.01a2.02 0.09a
Final body weight (g) 14.66 1.17b 4.63 0.03c 15.27 2.98b 19.69 1.21a
SGR2 3.50 0.13b 1.50 0.05c 3.55 0.34b 4.07 0.12a
FCR3 0.71 0.02b 1.41 0.06c 0.68 0.04ab 0.64 0.03a
PER4 2.50 0.06b 1.40 0.06b 2.98 0.18a 3.13 0.15a
NPU5 16.59 0.41b 7.30 0.41b 22.68 1.60a 23.79 2.01a
Final instantaneous food intake (% body weight/meal) 0.98 0.21b 0.50 0.33c 1.12 0.55b 1.88 0.22a
FCR 5 feed conversion ratio; NPU 5 net protein utilization; PER 5 protein efficiency ratio; SGR 5 specific growth rate.
1 Means of triplicate groups; values (6SD) with different superscript letters in the same row are significantly different

(P , 0.05).
2 SGR (%) 5 [(ln final weight ln initial weight) 3 100]/d.
3 FCR 5 dry feed intake (g)/wet body weight gain (g).
4 PER 5 body weight gain (g)/protein intake (g).
5 NPU (%) 5 [(final weight 3 final body crude protein) (initial weight 3 initial body crude protein) in (g)]/total
food 3 dietary crude protein in (g).
 EFFECTS OF CASEIN-BASED DIETS ON GROWTH, FEED EFFICIENCY OF RED PACU 251

FIGURE 2. Instantaneous feed intake of red pacu juveniles fed four experimental diets supplemented with 15% of wheat
meal (control), 15% of wheat camu-camu meal, 15% of aguaje meal, and 15% of maca meal for 8 wk. Values are
means 6 SD (n 5 3). Means annotated with different letters are significantly different (P , 0.05).

whole-body protein and ash levels were highest
in the camu-camu diet although protein levels
in experimental diets did not differ (Table 1;
P . 0.05). Fish fed the maca diet had signifi-
cantly higher K, Ca, Mg, and Zn concentrations
than fish fed the rest of the diets (P , 0.05;
Table 3).
All diets tested were isonitrogenous and met
the nutrient requirements in fish (NRC 1993;
Table 1). The macrominerals (calcium, phos-
phorus, potassium, and sodium) in the diets were
TABLE 3. Whole-body proximate and mineral composition of red belly pacu fed four experimental diets supplemented with
different Peruvian plants.
less than 1% of dry matter. The concentration of
trace minerals (copper, iron, magnesium, man-
ganese, and zinc) ranged from 4.3 to 168.5 mg/g.
Protein content ranged from 49.4 to 50.7%,
whereas ash comprised 4.44.8% of dry matter.
No histological pathologies were observed
in the stomach, intestine, pancreas, and pyloric
caeca of pacu fed the control diet or the diets
supplemented with different native plants tested
(Fig. 3). Normal appearance of the gastric
glands, lamina propria, muscularis, mucosal

Diets
Control Camu-camu Aguaje Maca
Proximate composition (%)
Protein 56.6 0.9bc 67.2 1.1a 54.4 0.8c 57.8 2.8b
Ash 11.1 0.1b 14.7 0.6a 9.8 0.1c 10.6 0.3b
Dry matter 24.8 0.6b 19.6 0.3c 26.6 1.4a 26.0 1.0ab
Mineral composition
Calcium (%) 2.68 0.02b 3.46 0.22a 2.27 0.07c 2.49 0.02bc
Phosphorus (%) 1.74 0.02b 2.30 0.11a 1.52 0.01d 1.63 0.01c
Potassium (%) 0.93 0.03b 1.28 0.04a 0.94 0.06b 0.96 0.04b
Sodium (%) 0.47 0.03b 0.62 0.01a 0.39 0.01c 0.40 0.04c
Magnesium (mg/g) 70 3b 130 5a 70 6b 70 2b
Zinc (mg/g) 10 1b 40 5a 10 2b 11 1b
Manganese (mg/g) 1 1a 2 1c 3 1b 3 0.1b
Means of triplicate groups; values (6SD) with different superscript letters in the same row are significantly different
(P , 0.05).
252 PALACIOS ET AL.

FIGURE 3. Cross-sections of the digestive system of red belly pacu (phosphate-buffered formalin; hematoxylin and eosin).
(A) Stomach showing gastric glands (g), lamina propria (l), and muscularis. (B) Intestine showing mucosal epithelium (me)
and goblet cells (gc). (C) Pancreas showing the pancreatic duct (pd), connective tissue (ct), acinar cells (ac), and fat cells
(fc) surrounding the pancreatic tissue. (D) Pyloric caeca showing cecal villi (cv), cecal wall (cw), and goblet cells (gc).

epithelium, goblet cells, pancreatic duct, con-
nective tissues, acinar cells, fat cells, cecal villi,
and cecal walls were observed. All the samples
from the initial sampling showed lipid infiltra-
tion and accumulation of brown, granular pig-
ments in the liver parenchyma. At the end of
this study, all the fish samples from the control
and camu-camu group and one of three samples
from the maca group had similar liver patholo-
gies noted at the beginning of the study. How-
ever, normal liver histology (polyhedral
hepatocytes with centrally located nuclei) was
observed in two of three samples from the maca
group and all the samples from the aguaje group.
Discussion
Present results with red pacu support previous
observations with rainbow trout that maca meal
supplementation into a semipurified diet in-
creases growth rate, feed efficiency, and protein
utilization (Dabrowski et al. 2003; Lee et al.
2004). The effects of maca on growth may
be attributed to its content of phytochemicals
(phytosterols and glucosinolates) and nutrient
characteristics (Dini et al. 1994; Obregon
1998; Canales et al. 2000; Cicero et al. 2001;
Li et al. 2001). Lee et al. (2004) attributed the
increased growth rate and survival in juvenile
rainbow trout to the presence of phytoestro-
gens (quercetin and isothiocyanates) that
may have crucial roles in growth and nutrient
utilization in fish. Phytochemicals in maca meal
have been reported to influence fertility,
immune resistance, and anabolic processes in
mammals (Gonzales et al. 2001; Sandoval
et al. 2002).
 EFFECTS OF CASEIN-BASED DIETS ON GROWTH, FEED EFFICIENCY OF RED PACU 253

FIGURE 4. Cross-sections of liver of red belly pacu (phosphate-buffered formalin; hematoxylin and eosin). (A) Well-
differentiated polyhedral hepatocytes with centrally located nuclei. (B) Accumulation of lipid vacuoles (lv) in the
parenchyma. (C) Magnified view of lipid vacuoles. (D) Accumulation of brown, granular pigments (pi) in the
parenchyma.

Maca meal may also contain certain attractants
that are responsible for the increased feed intake
in pacu (this study) and rainbow trout (Lee et al.
2004). Our study pioneered the use of aguaje and
camu-camu as dietary supplements in fish diets.
The notably high levels of ascorbic acid in camu-
camu (Moraes et al. 1994) open possibilities for
using this plant as a natural source of vitamin
C, as well as protein and minerals. The poor
growth of fish fed camu-camu-containing diet
cannot be associated with neutralization as it
improved initially feed acceptance. Previous
studies with pH neutralization of the diet demon-
strated an improved growth of channel catfish
(Wilson et al. 1977) and common carp (Nose
et al. 1974) when diets based on synthetic amino
acids were neutralized to pH 6. It suggests the
need to investigate potential antinutrients in this
fruit, camu-camu, palatability, and negative
effects on nutrient absorption in the digestive
tract of fish. The lack of ulcerations observed in
the gastric mucosa suggests that the acidity was
neutralized appropriately and did not affect
carbohydrate and protein digestion in the fish
fed camu-camu diets. It is not clear whether the
low growth in fish fed the camu-camu diet can
be attributed to the feed composition or mainly
to a reduced feed intake, or both. In this study,
the same protein level in diets resulted in signif-
icantly smaller fish size but statistically higher
final whole-body protein concentration in fish
fed the camu-camu diet (Tables 2, 3). These re-
sults suggest that protein is absorbed adequately
but not used as energy for growth. Further studies
254 PALACIOS ET AL.
on protein digestion and utilization in pacu fed
plant-based diets are warranted.
Our results have shown that the experimental
diets tested did not lead to pathologies in the
digestive tract of pacu juveniles. The presence
of lipid infiltration and pigment accumulation
in the liver parenchyma may be because of peri-
ods of low feed intake prior to the start of the
feeding experiment although the low number
of fish sampled for histological analyses limits
the validity of this observation (Fig. 4; see also
Storch and Juario 1983; Ferguson 1989). It
may also be possible that pacu has the tendency
to store large quantities of lipids in the liver.
Ostaszewska et al. (2005) did not observe any
pathologies in the digestive tract of pacu fed
caseingelatin-based diets supplemented with
soybean meal up to 50% crude protein. Cabal-
lero et al. (2004) observed steatosis (foci of
swollen hepatocytes containing numerous lipid
vacuoles) in sea bream fed with diets containing
vegetable oils (soybean oil, linseed oil, and
rapeseed oil) for 3 mo. The extent of lipid accu-
mulation in the liver was significantly reduced
when fish were refed with 100% fish oil, which
suggests that steatosis may be reversible. How-
ever, the presence of lipid infiltration in the liver
parenchyma of controls and experimental fish
in our study suggests a different mechanism of
action. The small number of samples collected
per treatment limits the validity of our results;
however, our study provides a general indication
of the physiological status of our experimental
fish. Further studies are needed to elucidate the
lipid accumulation and infiltration in the livers
of pacu juveniles.
In conclusion, maca meal has good potential
for use as supplements in caseingelatin-based
pacu diets. These diet formulations can be used
as a basis for further studies on nutrient require-
ments in this fish species. Results confirm previ-
ous studies (Lee et al. 2004) that maca meal may
be used in fish diets as a source of growth-pro-
moting substances and attractants to increase
diet palatability.
Acknowledgments
This research is a component of the Aquacul-
ture Collaborative Research Support Program
(CRSP), supported by USAID grant number
LAG-G-00-96-90015-00 and by contributions
from the participating institutions. The Aqua-
culture CRSP accession number is 1300. The
opinions expressed herein are those of the au-
thor(s) and do not necessarily reflect the views
of the U.S. Agency for International Develop-
ment. Maca meal was provided by Dr. Carlos
Gomez, Department of Nutrition, University of
Agriculture, La Molina, Lima, Peru.
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