ComP. Eiochem. Physiol.Vol. 103A, No. 2, pp. 413-415, 1992 0300-9629/92 $5.00 + 0.

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Printed in Great Britain 0 1992 Pergamon Press Ltd

BLOOD PARAMETERS OF THE AMAZONIAN MANATEE

(TRICHECHUS INUNGUIS)-DIETARY VARIATION

ELTON PINTO COLA=,* IONI GONCALVES-COLARES* and ARY DOMINGOSDO AMARAL~

*Institute National de Pesquisas de Amazonia, Caixa Postal 478, CEP 478, Manaus, Amazonas, Brasil;
tUniversidade de Slo Paulo, Instituto de Biociencias, Caixa Postal 11461, SZo Paulo, Brasil

(Received 14 January 1992)

Abstract-l. No significant differences (P > 0.05) were found between the blood parameters of male and
female Amazonian manatees.
2. When water hyacinth was introduced into the animals’ diet, there was a significant difference (P < 0.05)
in total lipid, total protein and albumin concentrations. However, there were no differences (P > 0.05) in
glucose and urea concentrations.
3. The total protein and albumin concentrations in the Amazonian manatee blood appear to be good
indicators for nutritional status, while urea appears not to be.

INTRODUCTION Blood was collected in the morning, 24 hr after the animals

Trichechus inunguis, the Amazonian manatee, is
endemic to the basin of the Amazon river and is the
only exclusively fresh water sirenian, being found
from the Island of Marajo, Brazil to the sources of
this river in Colombia, Peru and Equador (Domning,
1981; Timm et al., 1986). According to Best (1984),
its occurrence is restricted to calm waters and lakes
with abundant aquatic vegetation. This animal is
herbivorous, non-ruminant and feeds on a large
variety of aquatic plants. However, it has a preference
for aquatic grasses (Goncalves-Colares, 1991).
Nutrition plays a fundamental role in the physio-
logical state of the animals, and consequently, causes
changes in concentrations of blood parameters of the
individuals. The use of blood parameters as physio-
logical indices to determine the nutritional status of
wild animals has received a lot of attention. LeResche
et al. (1974) wrote a literature review on the vari-
ations in blood parameters of cervidae and correlated
these to the nutritional status of the animal. A similar
study was carried out by Warren et al. (1981)
studying the deer Odocoileus virginianus. Brown et al.
(1978) observed variations in the total protein and
urea content in the blood of the elephant Elephus
maximus, in relation to the food consumed at differ-
ent periods of the year. Therefore, the main objectives
of this project were to verify the possible variations
of some biochemical parameters in the blood of the
Amazonian manatee in relation to dietary changes, as
well as whether these parameters are related to the sex
of the animal.
MATERIALS AND METHODS
Nine adult Amazonian manatees (four male and five
female), which are kept captive at the National Institute of
Amazonian Research ‘(INPA) in a concrete tank with an area
of 78.5 m2 and a denth of 60 cm. were used. The animals have
been fed on para &ass (Brachi& mutica) since 1986, given
in sulIicient quantity to provide 5% of the animals’ live
weight per day.
were fed and always on the day that the pool was cleaned.
This is because the pool was emptied and thus reduced the
stress caused to the animals by handling. The animals were
immobilized using a restraining apparatus (Colares, 1991)
and the blood was withdrawn from the ventral side of the
pectoral flippers in the brachial artery sub-divisions. A
disposable 20 ml syringe was used with scalp needles, number
19 G. Ten millilitres of blood were removed from each
animal and separated between two tubes, one containing
heparin and the other an anti-coagulant for glucose.
Once the blood was removed, the tubes were quickly taken
to the laboratory where they were centrifuged at 15OOgfor
20 min. The plasma was stored at - 15°C. The biochemical
analysis of the plasmas were carried out, at the latest, 10 days
after being collected. Calorimetric analysis (kits LABTEST)
was carried out on these samples to determine the glucose
concentration by the enzymatic method (GOD-ANA), total
lipids, urea, albumin and total proteins. The globulin concen-
tration was calculated by the difference between the total
protein and albumin concentrations.
In order to find out whether the animals’ diet caused
alterations in the blood parameters, following blood collec-
tion from the nine animals, they were fed for a period of 28
days, during May 1990, on a diet consisting of 20% water
hyacinth (Eichhornia crassipes) and 80% para grass, con-
tributing a total of 5% live weight per day. The water
hyacinth was collected every day from Marchantaria Island,
near Manaus city (Sohmdes river, Amazon) in the morning,
and given to the animals on the same day. After this period,
blood was collected and processed as described earlier.
Eichhornia crussipes was used in this project because it is
easily found near the city of Manaus and because it is the
third most common plant in the manatees’ diet in the wild
(Goncalves-Colares, 1991). The fact that this plant contains
tannin, polyphenols and heavy metals was also taken into
consideration (Howard-Williams and Junk, 1977; Gopal and
Sharma, 198I; Wolter, personal communication), since they
decrease the absorption of proteins in the digestive tract of
the animals (Reese, 1979; Fraser and Mays, 1986; Hay and
Van Hoven, 1988).
During the experimental period, samples of the leaves
from the plants given to the animals were collected in order
to determine the percentages of crude protein, fats, ashes,
carbohydrates and humidity. For these analyses, the usual
analytical methods for nutritional plant constituents were
used (Silva, 1981; IBGE, 1981).

413
414 ELTON PINTOCOLARFSet al.

Table I. Comparative analyses (Student’s r-test) of the blood Table 2. Bromatological analyses of the plants
parameters between male and female Amazonian manatees fed on used in this study
oara arass and water hvacinth
Plants
Blood Male Female Brachiaria Eichhornia
parameters X S N X s N Sig mutica crassipes
W) (“10)
Glucose (mg/dl) 40.05 8.97 4 39.82 9.25 5 ns
Lipids (mg/dl) 582.91 115.48 4 603.26 147.92 5 ns Proteins 12.9 11.6 -
Urea (mg/dl) 22.93 3.33 4 29.32 12.68 5 ns Fat 6.5 3.1
~_,
Proteins feldl1
I
5.93 0.30 4 6.10 0.30 5 ns Carbohydrates 12.1 71.6
Albumin (g/dl) 2.88 0.19 4 2.84 0.18 5 ns Ashes 7.9 13.7
Globulins (g/dl) 3.05 0.26 4 3.26 0.33 5 ns Humidity 67.5 89.0
ns, Not significant; X, mean; S, standard deviation; N, number of
animals; Sig, level of significance.

In this study, the hypothesis test to compare means
(Student’s f-test) for equal variances was used, at a signifi-
cance level of 95%.
RESULTS AND DISCUSSION
The nine animals studied consumed all the water
hyacinth given to them and only the green leaves of
the para grass, leaving the stems and dried leaves. It
was, therefore, noted that the animals fed on 32%
water hyacinth and 68% para grass, contributing to
4.3% of their live weight per day. There was no
significant difference (P > 0.05) in the animals’
weights during the study.
The means and standard deviations of the glucose,
total lipid, urea, total protein, albumin and globulin
concentrations for the male and female Amazonian
manatees are given in Table 1. It was noted that there
was not any significant difference (P > 0.05) in the
concentrations of these substances between the sexes.
These results are similar to those reported by Moore
and Sikes (1967) who also did not find any significant
differences in the blood total lipid concentrations of
elephants of different sexes. The same thing was
observed by Medway et al. (1982) when the possible
protein variations in the blood of marine manatees
was studied.
The results of the bromatological analyses carried
out on the plants given to the animals during the
study are shown in Table 2. The percentages of ashes
and humidity in the water hyacinth were greater than
in the para grass, while the percentage of fat was
lower. The percentages of the crude proteins and
carbohydrates were similar between the two plants.
In Table 3 it can be seen that the introduction of
water hyacinth to the animals’ diet significantly re-
duced the total lipid, total protein and albumin
concentrations (P < 0.05). However, the glucose,
urea and globulin concentrations did not show any
significant difference (P > 0.05).
LeReche et al. (1974) and Brown and White
(1980) showed that cervidae and elephants, respect-
ively, fed on a high protein diet have higher
levels of urea in the blood than those fed on low
protein diets. Bahnak et al. (1979) suggested that
the urea concentration reflects the energy and/or
protein metabolism of cervidae. Kirkpatrick et al.
(1975) and Warren et al. (1982) suggest that high
protein and fat diets have opposite effects on
the concentration of blood urea in cervidae. There-
fore, urea is not always indicated as being a good
indicator for the nutritional status of animals. This
could have happened with the manatees since there
was not any variation in urea when the water hy-
acinth was added to their diet (Table 3). This is
because this plant inhibits the absorption of protein,
which could cause a decrease in the urea concen-
tration in the blood.
Kumar et al. (1972), Seal et al. (1978) and Warren
et al. (1981) confirm that the variations in the total
protein and albumin concentrations are not a good
index of nutritional status and that these only vary
when there is an enormous decrease in the crude
protein concentraton in the diet and also when this
situation is prolonged for a long period. These results
differ from those encountered in this study in that the
total protein and albumin concentrations (Table 3)
showed a significant decrease (P < 0.05) when water
hyacinth was added to the animals’ diet for a short
period (28 days). This indicates that these substances
may be a good index of nutritional status for the
Amazonian manatee.
The total lipid concentration was also significantly
lower (P < 0.05) when water hyacinth was added
to the manatees’ diet (Table 3). This decrease may
be due to the lower fat content consumed by the
animals, since water hyacinth contains half the
amount of fat compared to para grass (Table 2). Also,
the percentage of humidity in water hyacinth is much

Table 3. Comparative analyses (Student’s t-test) of the blood parameters between the
different diets
Blood Diet 1 Diet 2
parameters X S N X S N Sig
--
Glucose (mg/dl) 38.25 8.54 9 39.77 5.48 9 ns
Total lipids (mg/dl) 721.71 109.6 9 590.93 89.1 9 *
Urea (mg/dl) 24.11 6.24 9 23.37 2.41 9 ns
Total proteins (g/dl) 6.40 0.30 9 6.02 0.29 9 *
Albumin (gjdl) 3.46 0.21 9 2.86 0.17 9 *
Globulins (a/dlj 2.94 0.40 9 3.20 0.39 9 ns
ns, Not significant; Diet I, para grass; Diet 2, para grass + water hyacinth; X, mean; S,
standard deviation; N, number of animals; Sig, level of significance.
*Significant at P < 0.05.
 Blood parameters of th e Amazonian manatee
greater (89%) than in para grass (67.5%), thereby
decreasing even more the quantity of fats consumed.
The same was observed by Colares, Gonqalves-
Colares and Amaral (in preparation) who found
variations in the total fat contents in the manatees’
blood when the percentage of fat in the diet varied
throughout the year. Brown and White (1979) also
showed that when the consumption of fat of the
African elephant (Loxodonta africana) is varied, the
total lipid concentrations in the blood also varied.
Acknowledgemenrs-We thank Labtest Diagnostica Ltda
for donating the lab kits used in this study; Dr Adalberto
Val and MSc Mtrcia Cristina Caraciolo who loaded the
equipment necessary and the help of the veterinarian
Francisco Colares and all the employees of the Aquatic
Mammals Laboratory. The study was financed by the
Instituto National de Pesquisas de Amazonia.
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