Download effects of ethanol on brain and pancreas weights, serum sodium and

EFFECTS OF ETHANOL ON BRAIN AND PANCREAS WEIGHTS, SERUM SODIUM AND
POTASSIUM ALONG WITH HAEMATOLOGICAL PARAMETERS IN QUAILS (Coturnix
coturnix japonica)
Manshad Bashir and M. Tariq Javed
Department of Veterinary Pathology
University of Agriculture, Faisalabad-38040, Pakistan
ABSTRACT
The effects of ethanol were investigated in quails at different doses through drinking water.
One hundred and twenty Japanese quails of 39 days old were purchased from local market. In this
experiment 120 quails were randomly divided into five groups of 24 each (Groups 1, 2, 3, 4 and
5). They were offered ethanol at dose rates 2, 4, 8 and 16%, respectively, while group 5 served as
control. Quails offered 8 and 16% ethanol through drinking water showed dullness, depression and
staggering gait, while those given 2 and 4% ethanol showed decreased responsiveness for 2 to 3
hours. The brain weight and volume along with serum sodium and potassium decreased significantly
(P< 0.05) in all treated groups, while there was significant (P< 0.05) increase in relative weight of
the pancreas. Erythrocyte, packed cell volume, haemoglobin and leukocyte count increased
significantly (P< 0.05) in ethanol treated quails. The results of differential leukocyte count were
non-significant. The results suggests that ethanol at these dose levels has significant effects on
haematology, brain volume and serum electrolytes.
Key words:
ethanol, quails, serum, sodium, potassium, haematology, RBC, TLC, Hb, PCV, brain
weight, brain volume, pancreas weight.
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INTRODUCTION
Poultry farmers, especially those engaged in broiler production in Punjab Pakistan have
been observed to use ethanol in conditions of mild respiratory diseases and for growth promotion,
without knowing the effects on health and production of broilers. Ethanol is rapidly absorbed from
gastrointestinal tract which is faster during fasting state with peak blood levels attained within 30-60
minutes (Hagger &Forney, 1963). After the absorption, body begins to dispose it immediately and
liver is the major site of its metabolism. In the liver, first ethanol is converted into acetaldehyde
which is further converted into acetate and H-ions and then into carbon dioxide and water (Forney
& Hughes 1963). Ethanol has varied effects on many systems of the body (Klassen and Persaud,
1978) and long term ethanol usage affects the structure and function of the central nervous system
(Betz and Goldstein 1984). It suppresses certain brain functions that may lead to respiratory
depression and hypoxia. The toxic effects of ethanol depend on the amount of ethanol taken, time
of intake, genetics and the physical status of the individual (Klassen and Persaud, 1978). In very
large doses, it acts as a general anaesthetic (Eckardt, et al., 1998). Intragastric ethanol
administration (5g/kg) has been reported to cause brain injury in mice (Mansouri, et al., 2001).
Chronic exposure to ethanol can cause widespread cell loss in the brain, in some cases may cause
dementia in the mouse (Marinelli, et al., 2000), while ingestion of ethanol for short period has been
found responsible for decreased brain volume (Mann et al., 1992). In very high doses ethanol can
cause acute pancreatitis, loss of gross muscle control, hypotension, congestive heart failure and
sudden death. It has been reported that alcohol causes reduced pancreatic blood flow through
microcirculatory disturbance of the pancreas (Horie and Ishii, 2001). Further that excessive use of
ethanol, causes thickened blood with elevated haematocrit and decreased leukocyte count (Ramp et
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al., 1975). The chronic effects of ethanol are to promote isosmotic retention of water and
electrolytes due to increased ADH levels. Excess water and electrolytes are acutely excreted in
response to additional alcohol ingestion and it has been stated that alcoholic patients may have
electrolyte abnormalities (Regland, 1990). With these wide range of know effects of ethanol and it
use in broilers prompted to carry out the study for dissemination of results to farmers with
justification or to recommend its logical use in birds.
MATERIALS AND METHODS
Experiment was conducted to determine the effects of ethanol in quails. In this experiment
120 quails of 39 days of age were purchased from local market and were randomly divided into five
groups (1, 2, 3, 4 and 5) after four days to acclimatize. They were offered ethanol at dose rates 2,
4, 8 and 16%, respectively through drinking water, while group 5 served as control. Six birds from
each group were slaughtered at weekly interval for four weeks. They were slaughtered for the
collection of blood and organs (brain and pancreas). Blood samples of about 4 ml were collected
from each at the time of slaughtering for haematological studies and to obtain serum for electrolyte
study.
Brain and pancreas were collected and weighed to obtain relative weights of these organs.
Brain volume was also measured by placing the removed brain in known volume of Bouin’s solution
in a measuring cylinder and the difference of initial and final reading was recorded as volume of brain.
Tissue samples from these organs were preserved in Bouin’s solution for histopathological studies.
Haematology included determination of TEC, TLC, ESR, PCV, Hb and differential leukocyte
count (Benjamin, 1978). Serum electrolytes included determination of Na and K by flame photometric
method (Tennant et al., 1972).
Data thus obtained in the experiments were analyzed by using analysis of variance technique and
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means were compared by LSD test on personnel computer by using SAS 6.12 statistical software (SAS,
1996).
RESULTS AND DISCUSSIONS
Signs and Mortality
The signs observed in quails offered 8 and 16 % ethanol were depression, dullness, loss of
appetite and staggering gait, while in birds offered 2 and 4% were staggering gait for 2 to 3 hours,
general weakness and decreased responsiveness to feeding and watering. In rabbits, 1g/kg ethanol
has been reported to be sufficient to impair the performance and reversibly depresses the neurons
(Alexandrov et al. 2000) and the chronic exposure in mice has caused dementia (Marinelli et al.,
2000). During present study, mortality was observed only during 3rd week, one bird died of each
group received 8 and 16 % ethanol. Ethanol has been known to cause suppression of certain brain
functions that lead to respiratory depression and hypoxia (Klassen & Persaud, 1978) . Roselle and
Mendenhall, (1984) reported that infections and chronic liver injury were common cause of
morbidity and mortality in alcoholics. Thus it appears that quails at higher concentration of ethanol
can succumb to effects of ethanol, while can tolerate lower levels.
Effects on Live Weight, Brain and Pancreas
Effect of ethanol on live weight and pancreas weight of quails is shown in Table 1. During
1st, 3rd and 4th weeks, it decreased significantly (P< 0.05) in all treatment groups, while during
2nd week, it decreased (P< 0.05) in all groups except when given 2% ethanol. The lower weight
in ethanol offered birds was due to lower feed intake as were depressed (Unpublished data).
However, the weight of pancreas increased (P<0.05) in all treatment groups. However, earlier
study on ethanol (20%) at 2 ml per Kg dose, given thrice daily for 7 days between 21-28 days of
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age revealed no effect on relative pancreas weight in growth-selected, juvenile, meat-type chickens
(Peebles et al. 1996). However, pancreatitis has been reported in alcoholic patients (Lebedev,
1982) but was not observed in quails.
Effect of ethanol on brain weight and volume is shown in Table 1. During first four weeks,
relative weight of brain decreased significantly (P< 0.05) in all treatment groups. This indicates
effect of ethanol on brain in quails as well, which confirms earlier findings of Marinelli et al. (2000).
This effect of ethanol has earlier been related with neuronal death in somatosensorycortex due to
ethanol exposure (Mooney and Miller, 2001). During 1st week, brain volume increased in birds
given 2% ethanol, while decreased in birds given 4, 8 and 16% ethanol. During other weeks, it
decreased significantly (P< 0.05) in all treatment groups. The effects of ethanol on brain volume
may suggest loss of cells in brain tissue. However, both gross and light microscopic studies
revealed no lesion in these organs. Histological examination at light microscope level did not reveal
any detectable degenerative, necrotic or inflammatory changes.
Effects on Haematological parameters
Effects on erythrocytes count, PCV and haemoglobin are shown in Table 2. During
treatment period, erythrocyte count, PCV and haemoglobin increased significantly (P< 0.05) in all
treatment groups, which was in agreement with findings of Ramp et al., (1975) who reported that
due to excessive use of ethanol, blood becomes thick with elevated haematocrit and decreased
leukocyte count in rats and dogs. However, decrease in blood cellularity with a single large dose of
ethanol in rats (Kanwar & Tikoo, 1992) or with continuous use in this specie through bone marrow
depression has also been reported (Akingbami & Aire, 1994). While our results and of Ramp et al.
(1975) differed from those of Akingbami & Aire (1994) and Kanwar & Tikoo (1992). Acute loss of
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water and electrolytes due to alcohol ingestion as reported by Regland, (1990) was responsible in
causing relative elevation of all blood parameters.
Effects on total leucocytes count (TLC) and erythrocyte sedimentation rate (ESR) are
shown in Table 2. During treatment period, TLC increased significantly (P< 0.05) in all treatment
groups. The increase in leukocytes was also appeared to be relative rather than absolute because
of increased thickness of blood and loss of plasma. Our results however, differed from earlier
reports in rats or other animals with decrease in TLC (Kanwar and Tikoo, 1992; Ramp et al.,
1975). The results of ESR showed increase (P< 0.05) in birds offered 4 % or higher levels of
ethanol almost throughout the treatment period. This indicates effect if ethanol on settling ability of
RBC through some unknown mechanism.
Effects on Serum Sodium and Potassium
Effects on serum sodium and potassium are shown in Table 3.The results showed decrease
(P< 0.05) in serum sodium and potassium in birds of all groups almost throughout the treatment
trial. Ledig et al. (1985) reported that alcohol decreases Na - K ATPase activity that result in
decreased absorption of substances that required active, energy-dependent transport while
increased excretion of water and electrolytes in alcoholics has also been reported (Persson, 1991).
Fincham et al. (1986) also observed lower level of potassium in plasma of monkeys given ethanol
for long time. It can be inferred that effects of ethanol on serum electrolytes are significant (P<
0.05) causing reduction in their levels with possible reason of excretion of water and electrolytes in
response to ethanol ingestion (Regland, 1990) and the effect in birds are similar as in other birds.
Conclusions:
From the present study it can be concluded that ethanol at these dose levels has effects on
haematology, brain volume, serum electrolytes and performance in quails in particular which may be true
for other birds.
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Table 1:
Live weight, pancreas weight, brain weight and brain volume (Mean ± SD) in quails
given ethanol at various concentrations through drinking water
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