Effects of ethanol on brain and pancreas weights, serum sodium and potassium, and haematological parameters in quail (Coturnix coturnix japonica)

Abstract

Poultry farmers in Pakistan use ethanol for treatment of respiratory diseases and as a growth promotant, but the effects on health and production of broilers are unknown. The effects of ethanol were investigated in quail at different doses through drinking water. One hundred and twenty Japanese quail of 39 days of age were purchased from a local market. They were divided randomly into five groups of 24 (groups 1, 2, 3, 4 and 5) and offered ethanol at dose rates 2%, 4%, 8% and 16%, respectively, while group 5 served as control. Quail offered 8% and 16% ethanol showed dullness, depression and staggering gait, while those given 2% and 4% ethanol showed decreased responsiveness for 2 to 3 h. The live weight, brain 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. The decrease in live weight varied from 3% to 35% in a dose-related manner. The brain weight decreased by 4% to 10% and the decrease was dose dependent. Brain volume showed a 25% to 51% decrease. The haematological parameters erythrocyte count, packed cell volume and haemoglobin concentration increased significantly (P<0.05), while the leukocyte count increases were not significant. Both serum sodium and potassium decreased in ethanol-treated groups (36% to 44% and 44% to 49%, respectively), almost in a dose-dependent manner. The results suggest that ethanol at these dose levels has significant deleterious effects on haematology, brain volume and serum electrolytes.

Les éleveurs de volailles au Pakistan utilisent l'éthanol pour le traitement des maladies respiratoires et comme facteur de croissance, mais les effets sur la santé et la production des poulets de chair ne sont pas connus. Les effets de l'éthanol ont été étudiés chez la caille, par administration dans l'eau de boisson à différentes doses. Cent vingt cailles japonaises âgées de 39 jours ont été achetées sur un marché local. Elles ont été réparties au hasard en 5 groupes de 24 (groupes 1, 2, 3,4 et 5) et de l'éthanol a été mis à leur disposition à des taux respectifs de 2, 4, 8 et 16%, le groupe 5 a servi de control. Les cailles qui ont eu de l'éthanol à 8 et 16% ont montré un affaiblissement, de la dépression et une démarche titubante, alors que celles qui ont reçu 2 et 4% d'éthanol ont présentée une diminution de sensibilité pendant 2 à 3 heures. Le poids vif, le volume du cerveau ainsi que le sodium et le potassium sérique ont diminué significativement (P< 0,05) dans tous les groupes traités, alors qu'il y avait une augmentation significative (P< 0,05) du poids relatif du pancréas. La diminution du poids vif a varié de 3 à 35% en fonction de la dose. Le poids du cerveau a diminué de 4 à 10% et la diminution dépendait de la dose. Le volume du cerveau a montré 25 à 51% de diminution. Les paramètres hématologiques, le dénombrement des érythrocytes, le volume du culot cellulaire, et la concentration en hémoglobine ont augmenté significativement (P< 0,05) alors que les augmentations du nombre des leucocytes n'étaient pas significatives. Le sodium et le potassium ont diminué dans les groupes traités à l'éthanol (respectivement 36 à 44% et 44 à 49%), presque en fonction de la dose. Les résultats suggèrent que l'éthanol à ces niveaux de dose a des effets délétères significatifs sur l'hématologie, le volume du cerveau, et sur les électrolytes du sérum.

In Pakistan benutzen Geflügellandwirte Äthanol zur Behandlung von Respirations-erkrankungen und als Wachstumsförderer, aber die Auswirkungen auf die Gesundheit und die Produktion sind dabei unbekannt. Die Effekte von Äthanol auf Wachteln wurden bei verschiedenen Dosierungen über das Trinkwasser untersucht. 120 japanische Wachteln wurden im Alter von 39 Tagen auf einem lokalen Markt erworben. Sie wurden zufällig in fünf Gruppen à 24 (Gruppen 1, 2, 3, 4 und 5) aufgeteilt, denen 2, 4, 8 und 16%ige Äthanollösungen angeboten wurde. Die Gruppe 5 diente als Kontrolle. Die Wachteln, die 8 und 16 %iges Äthanol bekamen, zeigten Apathie, Depression und schwankenden Gang, während diejenigen, die 2 und 4%iges Äthanol erhielten, 2-3 Stunden lang ein vermindertes Reaktionsvermögen hatten. Das Lebendgewicht, das Gehirnvolumen und die Natrium- und Kaliumserumgehalte nahmen in allen behandelten Gruppen signifikant (p<0.05) ab, während das relative Pankreasgewicht signifikant (p<0.05) anstieg. Die Reduktion des Lebendgewichts variierte in Relation zur Dosis zwischen 3 und 35 %. Auch das Gehirngewicht reduzierte sich dosisabhängig um 4 bis 10 %. Das Gehirnvolumen nahm zwischen 25 und 51 % ab. Die hämatologischen Parameter Erythrozytenzahl, Hämatokrit und Hämoglobinkonzentration stiegen signifikant (p<0.05). Der Anstieg der Leukozytenzahl war dahingegen nicht signifikant. Sowohl die Natrium- als auch die Kaliumserumgehalte nahmen in den Äthanol behandelten Gruppen nahezu dosisabhängig ab (36 bis 44 % bzw. 44 bis 49 %) Die Ergebnisse deuten an, dass Äthanol in diesen Dosierungen signifikant schädliche Auswirkungen auf das Gehirnvolumen, hämatologische und Serumelektrolytwerte hat.

Los avicultores en Paquistán usan etanol para tratar las enfermedades respiratorias y como promotor del crecimiento, pero los efectos sobre la salud y producción de los pollos de engorde son desconocidos. Los efectos del etanol fueron estudiados en codornices a diferentes dosis administradas mediante agua de bebida. Ciento veinte codornices japonesas de 39 días de edad fueron adquiridas en un mercado local. Fueron divididas al azar en cinco grupos de 24 (Grupos 1, 2, 3, 4 y 5) y se les ofreció diferentes dosis de etanol, al 2, 4, 8 y 16%, respectivamente, mientras que el grupo 5 sirvió como control. Las codornices que tomaron 8 y 16% de etanol mostraron inactividad, depresión, y titubeo, mientras que las de 2 y 4% de etanol mostraron una respuesta retardada en 2 a 3 horas. El peso vivo, el volumen cerebral, así como el sodio y potasio séricos disminuyó significativamente (P< 0.05) en todos los grupos tratados, mientras que también se observó un incremento significativo (P< 0.05) del peso del páncreas. La disminución en el peso vivo varió entre el 3 y el 35% en relación a la dosis. El peso cerebral disminuyó entre el 4 y el 10% y la disminución fue dosis dependiente. El volumen cerebral mostró de un 25 a un 51% de disminución. Los parámetro hematológicos, contaje de eritrocitos, PCV y concentración de hemoglobina incrementaron significativamente (P< 0.05), mientras que los contajes de leucocitos no fueron significativos. Tanto el sodio como el potasio séricos disminuyeron en los grupos tratados con etanol (36 a 44% y 44 a 49%, respectivamente), casi de forma dependiente a la dosis. Los resultados sugieren que el etanol a estas dosis tienen efectos deletéreos significativos en hematología, volumen cerebral y electrolitos séricos.

Introduction

Poultry farmers, especially those engaged in broiler production in Punjab Pakistan, have used ethanol in treatment of conditions of mild respiratory diseases and for growth promotion, without knowing the effects on health and production of broilers. Ethanol is rapidly absorbed from the gastrointestinal tract, which is more rapid during the fasting state with peak blood levels attained within 30 to 60 min (Hagger & Forney, 1963). After absorption, the body begins to dispose of ethanol immediately, the liver being the major site of its metabolism. In the liver, ethanol is first converted into acetaldehyde, which is further converted into acetate and hydrogen ions and then into carbon dioxide and water (Forney & Hughes, 1963). Ethanol has varied effects on many systems of the body (Klassen & Persaud, 1978) and long-term ethanol usage affects the structure and function of the central nervous system (Betz & 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 & Persaud, 1978). In very large doses, it acts as a general anaesthetic (Eckardt et al., 1998). Intragastric ethanol administration (5 g/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, and in some cases may cause dementia in the mouse (Marinelli et al., 2000), while ingestion of ethanol for short periods has been found to be 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 & Ishii, 2001). Furthermore, it has been reported that excessive use of ethanol causes thickened blood with elevated haematocrit and decreased leukocyte count (Ramp et al., 1975). The chronic effects of ethanol are to promote isosmotic retention of water and electrolytes due to increased antidiuretic hormone 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). This wide range of known effects of ethanol, and its use in broilers, prompted the authors to carry out this study to determine any adverse effects in an avian model system using Japanese quail.

Materials and Methods

In this experiment, 120 quail of 39 days of age were purchased from local market and were randomly divided into five groups (groups 1, 2, 3, 4 and 5) after 4 days to acclimatize. They were given ethanol at dose rates 2%, 4%, 8% and 16%, respectively, through drinking water, while group 5 served as the control. These dose levels were selected keeping in view some of the previous studies on ethanol in the birds and laboratory animals as well as the use of ethanol (1% to 2% through drinking water) at commercial broiler farms in Pakistan. The higher dose levels were selected to determine any toxic effects in quails. Six birds from each group were killed by exsanguination (jugular vein) at weekly intervals for 4 weeks. The brain and pancreas were collected for further studies. Blood samples of about 4 ml were collected from wing vein prior to killing for haematological studies and to obtain serum for electrolyte study.

The 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 a known volume of Bouin's solution in a measuring cylinder and the difference between the initial and final readings recorded as the volume of the brain. Tissue samples from these organs were preserved in Bouin's solution for histopathological studies.

Haematology included determination of total erythrocyte count (TEC), total leukocyte count (TLC), erythrocyte sedimentation rate (ESR), packed cell volume (PCV), haemoglobin concentration and differential leukocyte count (Benjamin, 1978). Serum electrolytes included determination of sodium and potassium by the flame photometric method (Tennant et al., 1972).

Data thus obtained in the experiments were analysed using an analysis of variance technique and the means were compared by least significant difference test using SAS 6.12 statistical software (SAS Institute Inc., 1996).

Results and Discussions

Signs and mortality

The signs observed in quail that had been offered 8% and 16% ethanol were depression, dullness, loss of appetite and staggering gait, while in birds offered 2% and 4% the signs were staggering gait for 2 to 3 h, general weakness and decreased responsiveness to feeding and watering. In rabbits, 1 g/kg ethanol has been reported to be sufficient to impair performance and reversibly depresses the neurons (Alexandrov et al., 2000), and chronic exposure in mice has caused dementia (Marinelli et al., 2000). Similarly, an earlier study in wild birds reported an anaesthetic effect, causing them to fall from height, resulting in death (Fitzgerald et al., 1990). During the present study, mortality was observed only during the third week; one bird died in each group receiving 8% and 16% ethanol, respectively. Ethanol has been known to cause suppression of certain brain functions that lead to respiratory depression and hypoxia (Klassen & Persaud, 1978). Roselle & Mendenhall (1984) reported that infections and chronic liver injury were common causes of morbidity and mortality in alcoholics. Although ethanol has been reported to have toxic effects on liver and can cause death, these two quail did not reveal any gross or microscopic lesions in any of the organs including liver. The mortality in these birds may not have been associated with the ethanol, or ethanol might have some unknown role in this mortality.

Effects on live weight, brain and pancreas

The effect of ethanol on live weight and pancreas weight is presented in Table 1. During the first, third and fourth weeks live weight decreased significantly (P<0.05) in all treatment groups, while during the second week live weight decreased (P<0.05) in all groups except in those given 2% ethanol. However, at the end of fourth week, the decrease in live weight varied from 3% to 35% in groups offered 2% to 16% ethanol, respectively (Table 1). The lower live weight in ethanol-treated birds resulted from depressed feed intake. The total feed consumed per bird in each group (i.e. control, 2%, 4%, 8% and 16% ethanol) during the experiment was 701.82 g, 649.96 g, 623.54 g, 565.92 g and 489.92 g, respectively. However, the weight of pancreas increased (P<0.05) in all treatment groups by 0% to 56%, and the increase in pancreas weight was almost dose dependent with a difference in increase of 20%, particularly from 4% to 16% ethanol. An earlier study on ethanol (20%) at 2 ml/kg dose, given three times each day for 7 days between 21 and 28 days of age, revealed no effect on relative pancreas weight in growth-selected, juvenile, meat-type chickens (Peebles et al., 1996). Pancreatitis has also been reported in alcoholic patients (Lebedev, 1982), but was not observed in quail.

Table 1. Live weight, pancreas weight, brain weight and brain volume (mean±standard deviation) in quail given ethanol at various concentrations through drinking water

Group (% ethanol)	1 week	2 weeks	3 weeks	4 weeks	% Difference from control
Live weight (g)
Group 1 (2%)	145.00±4.08	153.50±4.20	158.18±2.31*	179.84±2.60*	03% decrease
Group 2 (4%)	131.25±6.29*	135.21±2.60*	132.25±5.20*	133.71±0.73*	27% decrease
Group 3 (8%)	128.75±6.29*	134.49±0.99*	129.09±2.56*	130.79±0.79*	29% decrease
Group 4 (16%)	127.50±6.45	134.92 ±0.57*	110.62±3.09*	119.50±3.42*	35% decrease
Group 5 (0%)	150.00±7.07	153.38±2.61	178.81±2.63	185.70±3.88
Pancreas weight (g)
Group 1 (2%)	0.29±0.00*	0.29±0.00*	0.30±0.00	0.30±0.01	00% increase
Group 2 (4%)	0.30±0.01*	0.31±0.01*	0.33±0.00*	0.35±0.00*	16% increase
Group 3 (8%)	0.32±0.00*	0.34±0.01*	0.36±0.01*	0.41±0.01*	36% increase
Group 4 (16%)	0.34±0.01*	0.37±0.00*	0.44±0.01*	0.47±0.01*	56% increase
Group 5 (0%)	0.27±0.00	0.28±0.00	0.29±0.01	0.30±0.01
Brain weight (g)
Group 1 (2%)	0.51±0.01*	0.50±0.01*	0.49±0.00*	0.47±0.01	04% decrease
Group 2 (4%)	0.50±0.00*	0.49±0.01*	0.47±0.00*	0.46±0.00	06% decrease
Group 3 (8%)	0.50±0.00*	0.48±0.00*	0.46±0.00*	0.45±0.00	08% decrease
Group 4 (16%)	0.50±0.00*	0.47±0.00*	0.45±0.00*	0.44±0.00*	10% decrease
Group 5 (0%)	0.55±0.01	0.54±0.01	0.50±0.00	0.49±0.05
Brain volume (ml)
Group 1 (2%)	0.46±0.06	0.43±0.01*	0.42±0.01*	0.40±0.02*	25% decrease
Group 2 (4%)	0.42±0.04	0.35±0.00*	0.34±0.01*	0.33±0.02*	38% decrease
Group 3 (8%)	0.43±0.03	0.32±0.01*	0.32±0.02*	0.30±0.01*	44% decrease
Group 4 (16%)	0.41±0.01	0.32±0.03*	0.30±0.01*	0.26±0.01*	51% decrease
Group 5 (0%)	0.45±0.01	0.54±0.03	0.53±0.03	0.54±0.04
*Significant difference (P<0.05) compared with control group.

The effect of ethanol on brain weight and volume is presented in Table 1. During the first 3 weeks, the relative weight of brain decreased significantly (P<0.05) while at the fourth week it decreased non-significantly in all treatment groups except those given 16% ethanol. The decrease in brain weight varied from 4% to 10%, with a difference of 2% at increasing dose level. This indicates an effect of ethanol on the brain in quail, which confirms earlier findings of Marinelli et al. (2000). This effect had earlier been related to neuronal death in the somatosensory cortex (Mooney & Miller, 2001). During the first week the brain volume differed non-significantly, while during other weeks it decreased significantly (P<0.05) in all treatment groups, with a decrease of 25% to 51% with increases in ethanol level. The difference in decrease in brain volume was 14% at 2% to 4% ethanol, was 6% at 4% to 8% ethanol and was 7% at 8% to 16% ethanol. The effects of ethanol on brain volume may suggest loss of cells or cell volume in brain tissue. However, gross examination revealed no lesion in the brain. Histological examination also failed to reveal any detectable degenerative, necrotic or inflammatory changes. The results of brain weight and brain volume showed a slightly greater decrease of volume compared with weight at 2% to 16% ethanol levels, while the decrease in brain volume was drastic from 2% to 4% ethanol compared with a decrease in brain weight at these ethanol levels.

Effects on haematological parameters

Effects on TEC, PCV, haemoglobin concentration, TLC and ESR are presented in Table 2. During the treatment period, TEC, PCV, haemoglobin concentration, and TLC increased significantly (P<0.05) in all treatment groups. This was in partial agreement with findings of Ramp et al. (1975), who reported that due to excessive use of ethanol, blood became thick with elevated haematocrit and decreased leukocyte count in rats and dogs. However, a decrease in blood cellularity with a single large dose of ethanol in rats (Kanwar & Tikoo, 1992) or with continuous use in this species through bone marrow depression has also been reported (Akingbemi & Aire, 1994). Our results and those of Ramp et al. (1975) differed from those of Akingbemi & Aire (1994) and Kanwar & Tikoo (1992). Acute loss of water and electrolytes due to alcohol ingestion as reported by Regland (1990) may be the possible reason of relative elevation of all the blood parameters. The increases in TEC, PCV, haemoglobin concentration were in the ranges 95% to 189%, 32% to 54% and 52% to 71%, respectively. However, the difference between treatments in TEC was 76% at dose 2% to 4%, was 12% at dose 4% to 8% and was 6% at dose 8% to 16%. Such differences in PCV were 7%, 10% and 5%, in haemoglobin were 3, 3 and 5, while in TLC were 13%, 38% and 25% at an ethanol dose of 2% to 4%, of 4% to 8% and of 8% to 16%, respectively. The ESR increased by 115% to 172% with increase in ethanol level (Table 2), with a difference of 46%, 11% and 0% at an ethanol dose of 2% to 4%, of 4% to 8% and of 8% to 16%, respectively. This shows a slightly higher increase in TEC, TLC and ESR, with a greater difference between doses compared with PCV and haemoglobin concentration. It may be mentioned that during the present study no inflammatory or reparative process could be observed in the studied tissues but bone marrow examination was not performed. Our results, however, differed from earlier reports in rats or other animals with decrease in TLC (Ramp et al., 1975; Kanwar & Tikoo, 1992).

Table 2. Erythrocyte count, PCV, haemoglobin, TLC and ESR (mean±standard deviation) in quail given ethanol at various concentrations through drinking water

Group (% ethanol)	1 week	2 weeks	3 weeks	4 weeks	% Difference from control
Erythrocyte count (10^6/μl)
Group 1 (2%)	2.38±0.08*	2.71±0.13*	3.08±0.52*	2.88±0.07*	95% increase
Group 2 (4%)	3.63±0.03*	3.67±0.06*	3.83±0.22*	3.99±0.04*	171% increase
Group 3 (8%)	3.86±0.08*	3.94±0.05*	4.11±0.06*	4.17±0.04*	183% increase
Group 4 (16%)	4.00±0.07*	34.06±0.09*	4.20±0.04*	4.26±0.04*	189% increase
Group 5 (0%)	1.30±0.02	1.38±0.03	1.46±0.07	1.47±0.05
PCV (%)
Group 1 (2%)	34.00±0.82*	36.50±3.11*	36.75±2.50*	40.00±0.82*	32% increase
Group 2 (4%)	34.25±0.96*	36.25±1.50*	40.00±0.82*	42.00±1.41*	39% increase
Group 3 (8%)	38.50±1.91*	39.00±1.73*	41.25±1.50*	45.00±0.82*	49% increase
Group 4 (16%)	39.75±1.50*	40.33±2.08*	44.75±0.96*	46.75±1.50*	54% increase
Group 5 (0%)	27.50±1.91	28.00±3.27	28.25±3.20	30.20±2.17
Haemoglobin (g/dl)
Group 1 (2%)	11.73±1.03*	12.63±0.68*	13.30±1.30*	13.63±0.17*	52% increase
Group 2 (4%)	12.23±1.18*	13.21±1.30*	13.71±1.10*	13.71±1.10*	53% increase
Group 3 (8%)	12.48±1.02*	13.46±1.09*	14.04±1.10*	13.96±0.32*	56% increase
Group 4 (16%)	13.13±0.99*	13.33±1.25*	13.96±0.63*	15.28±0.87*	71% increase
Group 5 (0%)	9.50±0.32	9.09±1.13	8.84±0.87	8.92±1.23
TLC (10^3/μl)
Group 1 (2%)	56.50±2.65*	61.75±2.50*	69.50±2.08*	76.50±1.91*	155% increase
Group 2 (4%)	60.00±0.82*	65.25±1.71*	75.50±2.52*	80.50±1.73*	168% increase
Group 3 (8%)	61.50±1.91*	67.67±1.15*	81.75±2.22*	92.00±2.94*	206% increase
Group 4 (16%)	64.25±2.50*	69.67±0.58*	84.75±3.95*	99.50±3.32*	231% increase
Group 5 (0%)	28.50±1.91	28.50±1.30	29.50±2.89	30.00±1.41
ESR (mm/h)
Group 1 (2%)	3.25±0.50	3.75±0.50	4.00±1.15	4.75±1.50*	115% increase
Group 2 (4%)	3.50±0.58*	4.25±0.50*	4.25±1.26	5.75±0.50*	161% increase
Group 3 (8%)	4.50±0.58*	4.67±1.15*	4.75±0.96	6.00±0.82*	172% increase
Group 4 (16%)	4.75±0.96*	5.00±1.00*	4.75±0.96	6.00±0.82*	172% increase
Group 5 (0%)	2.00±0.82	2.50±0.58	2.75±0.96	2.20±0.84
*Significant difference (P<0.05) compared with control group.

Effects on serum sodium and potassium

Effects on serum sodium and potassium are presented in Table 3.The results showed decreases (P<0.05) in serum sodium and potassium in birds of all groups almost throughout the treatment trial. The decrease in serum sodium was drastic in quail offered 2% to 4% (5% difference) ethanol, moderate from 4% to 8% ethanol (2% difference) and mild from 8% to 16% ethanol (1% difference). However, potassium levels decreased drastically in quail offered 2% to 4% ethanol, while no change in potassium levels observed in quail offered 4% to 16% ethanol. Ledig et al. (1985) reported that alcohol decreases Na-K-ATPase activity that results 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 levels of potassium in the 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, possibly due to excretion of water and electrolytes in response to ethanol ingestion (Regland, 1990), and the effect in these birds is similar to that reported in other birds.

Table 3. Serum sodium and potassium (mean±standard deviation) of quail given ethanol at various concentrations through drinking water

Group (% ethanol)	1 week	2 weeks	3 weeks	4 weeks	% Difference from control
Sodium
Group 1 (2%)	115.54±3.06*	107.59±4.33*	98.32±2.65*	96.99±4.33*	36% decrease
Group 2 (4%)	107.59±4.33*	96.99±4.33*	94.34±3.06*	90.37±2.65*	41% decrease
Group 3 (8%)	94.34±3.06*	93.46±3.06*	90.37±2.65*	86.39±4.33*	43% decrease
Group 4 (16%)	86.72±5.07*	95.22±3.06*	90.37±5.07*	85.07±5.07*	44% decrease
Group 5 (0%)	148.67±5.07	146.02±9.05	147.34±9.18	153.17±2.12
Potassium
Group 1 (2%)	11.14±2.63	9.08±2.25*	9.08±2.25*	8.39±1.38*	44% decreased
Group 2 (4%)	10.45±2.75	9.76±2.63*	9.08±2.25*	7.70±1.59*	49% decrease
Group 3 (8%)	7.70±1.59*	9.99±1.59*	9.08±2.25*	7.70±1.59*	49% decrease
Group 4 (16%)	8.39±1.38*	8.16±3.18*	9.76±1.38*	7.70±1.59*	49% decrease
Group 5 (0%)	13.20±2.75	15.95±2.75	14.58±2.25	15.13±1.60
*Significant difference (P<0.05) compared with control groups.

It can be concluded from our study that ethanol at these dose levels has caused significant increase in haematological parameters, with significant decrease in brain volume and serum electrolytes, the effects being almost invariably dose dependent. Among serum electrolytes studied, sodium showed a dose-dependent response while potassium did not. It is further emphasized that ethanol has serious effects on quails. Therefore, broiler farmers are advised not to use ethanol for any purpose.

Translations of the abstract in French, German and Spanish are available on the Avian Pathology website.