IV. RESULTS
4.1 Ultraviolet spectroscopy
4.1.1 Ultraviolet visible spectrum of diclofenac sodium
The ultraviolet visible spectrum (200 – 350 nm) of diclofenac sodium in acidic
(0.1 N HCl), alkaline (0.1 N NaOH) and neutral (methanol) solvent systems are shown in
Fig. 1. The maximum ultraviolet absorption of diclofenac was found at 273, 275, 279 nm
in acidic (0.1 N HCl), alkaline (0.1 N NaOH) and neutral (methanol) solvent systems,
respectively. The obtained spectra and maximum absorption wavelength for diclofenac
sodium in acidic and alkaline solvent system was compared with the reference spectra of
diclofenac sodium.
2.0
Absorbance
1.5
Acidic Solvent
Alkaline Solvent
Neutral Solvent
279
275
1.0
273
0.5
0.0
200
225
250
275
300
Wavelength in nm
Fig. 1. UV-visible spectrum of diclofenac sodium
325
350
61
4.1.2 Ultraviolet visible spectrum of aspirin
The ultraviolet visible spectrum (200 - 350 nm) of aspirin in acidic (0.1 N HCl),
and neutral (methanol) solvent systems are shown in Fig. 2. The maximum ultraviolet
absorption of aspirin was found at 230 and 278 nm in acidic (0.1 N HCl) and 225 and 276
nm in neutral (methanol) solvent systems, respectively. The obtained spectra and
maximum absorption wavelength for aspirin in acidic solvent system was compared with
the reference spectra of aspirin.
1.5
230
Absorbance
1.0
Acidic Solvent
Neutral Solvent
225
0.5
278
276
0.0
200
225
250
Wavelength in nm
Fig. 2. UV-visible spectrum of aspirin
275
300
62
4.1.3 Ultraviolet visible spectrum of paracetamol
The ultraviolet visible spectrum (200 - 325 nm) of parcetamol in acidic (0.1 N
HCl), alkaline (0.1 N NaOH) and neutral (methanol) solvent systems are shown in Fig. 3.
The maximum ultraviolet absorption of paracetamol was found at 245, 256 and 250 nm
in acidic (0.1 N HCl), alkaline (0.1 N NaOH) and neutral (methanol) solvent systems,
respectively. The obtained spectra and maximum absorption wavelength for paracetamol
in acidic and alkaline solvent system was compared with the reference spectra of
paracetamol.
2.0
250
Acidic Solvent
Alkaline Solvent
Neutral Solvent
Absorbance
1.5
245 256
1.0
0.5
0.0
200
225
250
275
Wavelength in nm
Fig. 3. UV- visible spectrum of paracetamol
300
325
63
4.1.4 Ultraviolet visible spectrum of nimesulide
The ultraviolet visible spectrum (200 - 550 nm) of nimesulide in acidic (0.1 N
HCl), alkaline (0.1 N NaOH) and neutral (methanol) solvent systems are shown in Fig. 4.
The maximum ultraviolet absorption of nimesulide was found at 301, 394 and 400 nm in
acidic (0.1 N HCl), alkaline (0.1 N NaOH) and neutral (methanol) solvent systems,
respectively. The obtained spectra and maximum absorption wavelength for nimesulide
in acidic and alkaline solvent system was compared with the reference spectra of
nimesulide.
2.0
1.5
Absorbance
Acidic Solvent
Alkaline Solvent
Neutral Solvent
400
301
394
1.0
0.5
0.0
200
250
300
350
400
Wavelength in nm
Fig. 4. UV- visible spectrum of nimesulide
450
500
550
64
4.1.5 Ultraviolet visible spectrum of ketoprofen
The ultraviolet visible spectrum (200 - 350 nm) of ketoprofen in acidic (0.1 N
HCl), alkaline (0.1 N NaOH) and neutral (methanol) solvent systems are shown in Fig. 5.
The maximum ultraviolet absorption of ketoprofen was found at 260, 262 and 257 nm in
acidic (0.1 N HCl), alkaline (0.1 N NaOH) and neutral (methanol) solvent systems,
respectively. The obtained spectra and maximum absorption wavelength for ketoprofen
in acidic and alkaline solvent system was compared with the reference spectra of
ketoprofen.
2.0
262
257
260
Acidic Solvent
Alkaline Solvent
Neutral Solvent
Absorbance
1.5
1.0
0.5
0.0
200
250
300
Wavelength in nm
Fig. 5. UV- visible spectrum of ketoprofen
350
65
4.1.6 Ultraviolet visible spectrum of meloxicam
The ultraviolet visible spectrum (200 - 400 nm) of meloxicam in acidic (0.1 N
HCl), alkaline (0.1 N NaOH) and neutral (methanol) solvent systems are shown in Fig. 6.
The maximum ultraviolet absorption of meloxicam was found at 345, 362 and 356 nm in
acidic (0.1 N HCl), alkaline (0.1 N NaOH) and neutral (methanol) solvent systems,
respectively. The obtained spectra and maximum absorption wavelength for meloxicam
in acidic and alkaline solvent system was compared with the reference spectra of
meloxicam.
2.0
362
Acidic Solvent
Alkaline Solvent
Neutral Solvent
Absorbance
1.5
356
1.0
345
0.5
0.0
200
250
300
350
Wavelength in nm
Fig. 6. UV- visible spectrum of meloxicam
400
66
4.1.6 Ultraviolet visible spectrum of celecoxib
The ultraviolet visible spectrum (200 - 350 nm) of celecoxib in acidic (0.1 N
HCl), alkaline (0.1 N NaOH) and neutral (methanol) solvent systems are shown in Fig. 7.
The maximum ultraviolet absorption of celecoxib was found at 250, 252 and 254 nm in
acidic (0.1 N HCl), alkaline (0.1 N NaOH) and neutral (methanol) solvent systems,
respectively. The obtained spectra and maximum absorption wavelength for celecoxib in
acidic and alkaline solvent system was compared with the reference spectra of celecoxib.
254
2.0
250
1.5
Absorbance
Acidic Solvent
Alkaline Solvent
Neutral Solvent
252
1.0
0.5
0.0
200
225
250
275
300
Wavelength in nm
Fig. 7. UV- visible spectrum of celecoxib
325
350
67
4.2 Infrared spectroscopy
4.2.1 Infrared spectrum of diclofenac sodium
The infrared spectrum of diclofenac sodium in KBr and ATR is shown in Fig. 8
and Fig. 9, respectively. The principal wave numbers obtained in infrared spectrum and
their corresponding assignment (bond, compound type and functional groups) were
characteristic for diclofenac sodium as mentioned below.
Wave number
(cm-1)
KBr disk
Wave number
(cm-1)
ATR
Assignment of wave numbers
3387.39
3225.85
N-H stretching
1283.85, 1305.78
1282.16, 1305.08
C-N stretching 20 aromatic
1089.14
1089.14
C-X stretching (X=Chloride)
-
1044.77, 1089.19
C-H in plane bending aromatic
1194.25, 1166.11
1234.26, 1250.09
1282.16
769.40, 747.72
714.78, 744.82
C-H out of plane bending-odisubstituted
1400-1500 &
1585-1600
1451.29, 1604.01
C=C stretching aromatic
10
4000
3500
3000
2500
Wavenumbers (cm-1)
2000
Fig. 9. Infrared spectrum of diclofenac sodium (ATR)
1500
1000
484.25
714.78
452.62
633.60
840.00
1000
530.31
558.46
868.87
1044.77
70
1089.19
1500
951.92
60
1194.25
1393.30
40
769.40 747.72
715.71
869.26 839.68
1045.01
1199.65
1089.14
1249.39
952.02
1305.78 1283.85
1453.18
60
744.82
40
1166.11
1508.27
1470.43
50
1234.26
1250.09
2000
1384.39
1555.54
1574.72
%T
70
766.37
20
1469.21
2500
1451.29
3000
1553.24
3500
1497.41
20
1282.16
1305.08
30
1604.01
50
3225.86
4000
3387.39
30
1572.80
%T
68
100
90
80
10
-0
Wavenumbers (cm-1)
500
Fig. 8. Infrared spectrum of diclofenac sodium (KBr disk)
100
90
80
-0
500
69
4.2.2 Infrared spectrum of aspirin
The infrared spectrum of aspirin in KBr and ATR is shown in Fig. 10 and Fig. 11,
respectively. The principal wave numbers obtained in infrared spectrum and their
corresponding assignment (bond, compound type and functional group) were
characteristic for aspirin.
Wave number
(cm-1)
KBr disk
Wave number
(cm-1)
ATR
Assignment of wave numbers
1753.61
1748.91
C=O stretching- carboxylic acids
1220.82, 1306.84
1011.70, 1092.97,
C-O stretching - esters
1218.24, 1286.83
1306.84
1286.83
C-O stretching – esters of aromatic acids
2699.86, 2921.91
2829.67
O-H stretching carboxylic acids
1013.08, 1038.88,
1011.70, 1092.97,
C-H in plane bending aromatic
1094.86, 1134.99,
1218.24, 1286.83
1187.80, 1220.82
755.52
802.42, 838.38
C-H out of plane bending-o-disubstituted
1419.59, 1458.02,
1483.60
1445.74, 1481.99
C=C stretching aromatic
4000
3500
3000
2500
Fig. 11. Infrared spectrum of aspirin (ATR)
50
Wavenumbers (cm-1)
2000
1500
1000
514.31
665.78
423.40
643.16
802.42
1000
541.83
597.22
838.38
60
1011.70
1500
914.33
70
1092.97
2000
969.84
2500
1417.81
3000
790.71
970.41
1094.86
1187.80
840.28
1306.84
1220.82
1370.62
755.52
917.32
1038.88
1483.60
1605.77
704.84
1013.08
1134.99
1419.59
35
1218.24
15
1458.02
1753.61
25
1604.38
20
1691.97
30
1286.83
3500
666.72
804.09
1575.95
2699.86
2921.91
40
1367.98
65
1748.91
%T
45
1455.74
55
1680.72
80
2829.67
4000
1481.99
%T
70
60
55
50
10
Wavenumbers (cm-1)
500
Fig. 10. Infrared spectrum of aspirin (KBr disk)
90
85
75
500
71
4.2.3 Infrared spectrum of parcetamol
The infrared spectrum of paracetamol in KBr disk and ATR is shown in Fig. 12
and Fig. 13, respectively. The principal wave numbers obtained in infrared spectrum and
their corresponding assignment (bond, compound type and functional group) were
characteristic for paracetamol. The characteristic wave numbers and their corresponding
assignment are mentioned below.
Wave number
(cm-1)
KBr disk
Wave number
(cm-1)
ATR
Assignment of wave numbers
3325.87
3318.96
OH stretching phenolics
1655.79
1650.57
N-H bending
1226.16,1259.81,
1224.43,
C-O stretching phenolics
1370.53
1258.16, 1370.52
1015.16, 107.71,
1015, 1107.37,
1172.00, 226.61,
1171.56,
1243.18, 259.81
1224.43, 1258.16
808.30, 837.15
806.49, 837.78
C-H out of plane bending-p-disubstituted
1442
1434.05
C=C stretching aromatic
C-H in plane bending aromatic
4000
3500
3000
2500
Fig. 13. Infrared spectrum of paracetamol (ATR)
35
30
25
Wavenumbers (cm-1)
2000
40
1500
1000
501.26
602.83
464.21
414.32
967.91
1000
517.48
624.42
795.82
50
680.88
711.92
60
806.49
1500
835.78
2000
1171.56
65
2500
857.31
55
1327.64
857.12
1243.18
1259.81
808.30
1226.61
1370.53
837.15
713.94
685.11
968.47
1107.71
796.28
1015.16
1172.00
1327.37
58
1107.37
70
3000
1506.40
60
1224.43
44
1442.17
62
1015.08
45
1370.52
48
1258.16
3500
1610.59
50
1564.74
64
1434.05
46
1655.79
66
2586.72
70
1505.10
68
2793.21
1877.20
72
1561.93
52
3161.85
56
3109.19
54
3325.87
%T
74
1609.71
75
3318.96
4000
1650.57
%T
72
78
76
42
40
Wavenumbers (cm-1)
500
Fig. 12. Infrared spectrum of paracetamol (KBr disk)
100
95
90
85
80
20
500
73
4.2.4 Infrared spectrum of nimesulide
The infrared spectrum of nimesulide in KBr disk and ATR is shown in Fig. 14
and Fig. 15, respectively. The principal wave numbers obtained in infrared spectrum and
their corresponding assignment (bond, compound type and functional group) were
characteristic for nimesulide.
Wave number
(cm-1)
KBr disk
Wave number
(cm-1)
ATR
Assignment of wave numbers
1343.79, 1152.0
-
S=O stretching - Sulfonamides
3281.83
3277.32
N-H stretching - Sulfonamides (10)
740.75, 753.23,
802.46, 830.15
N-O stretching - Nitrites
871.85
871.08
C-N stretching - Nitroaromatics
1018.99,1126.84,
1215.49, 1245.50
C-H in plane bending - Aromatics
740.75, 803.80
696.85, 802.46
C-H out of plane bending-m-disubstituted
1487.87,1599.48,
1404.34, 1445.31,
C=C stretching - Aromatic
1588.51
1486.66, 1587.56
803.80, 830.71,
848.69
1068.91,1080.06,
1217.48,1152.07,
1185.92,1247.91,
1282.18
3500
3000
2500
Fig. 15. Infrared spectrum of nimesulide (ATR)
40
35
30
25
Wavenumbers (cm-1)
2000
1500
1000
20
500
445.10
1000
478.56
1185.97
950.64
976.17
830.71
1247.91
1599.48
803.80
905.42
753.23
871.85
1018.99
848.69
1735.33
698.07
1406.41
740.75
1217.48
1126.84
1068.91
1445.31
1080.06
1152.07
1282.18
1316.31
1343.79
1487.87
1502.90
2928.91
1700.97
663.76
72
512.97
551.19
740.03
802.46
1560.30
66
696.85
830.15
50
1078.95
65
1500
871.08
2000
903.71
2500
1215.49
75
3000
949.73
3500
1404.34
56
1522.45
60
1653.90
62
972.98
55
1445.31
80
3085.39
70
1149.50
45
1314.51
58
1588.51
64
3281.83
3438.86
3727.44
74
1245.50
70
1587.56
4000
3277.32
%T
68
1486.66
%T
74
76
54
Wavenumbers (cm-1)
500
Fig. 14. Infrared spectrum of nimesulide (KBr disk)
100
95
90
85
60
75
4.2.5 Infrared spectrum of ketoprofen
The infrared spectrum of ketoprofen in KBr disk and ATR is shown in Fig. 16 and
Fig. 17, respectively. The principal wave numbers obtained in infrared spectrum and their
corresponding assignment (bond, compound type and functional group) were
characteristic for ketoprofen.
Wave number
(cm-1)
KBr disk
Wave number
(cm-1)
ATR
Assignment of wave numbers
1697.38
1654.50
C=O stretching ketones
1134.57, 1174.86,
1134.14, 1174.57,
C=O stretching and bending
1194.79, 1227.84,
1195.28, 1227.32,
1285.34
1282.64
2978.67
2978.64
O-H stretching carboxylic acid
1420
1420.61
O-H bending carboxylic acid
1284.34, 1319.28
1318.92, 1282.64
C-O stretching phenolics
1078.31, 1134.57,
1077.65, 1134.14,
C-H in plane bending - Aromatic
1174.86, 1194.79,
1174.57, 1195.28,
1227.84, 1285.34.
1227.32, 1282.64
691.02, 703.53,
689.94, 703.13,
716.75, 773.10,
714.77, 787.17,
787.29
773.13
1445.31, 1420.60,
1420.61, 1441.26,
1598.20
1597.68
C-H out of plane bending-mdisubstituted
C=C stretching aromatic
4000
35
30
3500
3000
2500
Fig. 17. Infrared spectrum of ketoprofen (ATR)
45
Wavenumbers (cm-1)
2000
1500
1000
642.07
811.27
966.26
50
689.94
55
826.69
613.68
787.17
915.92
65
440.06
773.13
1077.65
80
513.66
716.75
866.07
1227.84
1285.34
703.53
826.75
1194.79
1420.60
691.02
811.17
968.08
1134.57
1369.88
1598.20
787.29
915.97
1078.31
1174.86
1575.56
773.10
60
703.13
40
1500
714.77
70
1319.28
30
1174.57
2000
1134.14
2500
1195.28
3000
1369.52
35
1227.32
60
1420.61
25
1445.31
50
1282.64
1318.92
3500
1597.68
20
1697.38
40
1655.21
45
2978.67
%T
55
1441.26
75
2978.64
4000
1654.50
%T
76
70
65
15
Wavenumbers (cm-1)
1000
500
Fig. 16. Infrared spectrum of ketoprofen (KBr disk)
90
85
500
77
4.2.6 Infrared spectrum of meloxicam
The infrared spectrum of meloxicam in KBr disk and ATR is shown in Fig. 18
and Fig. 19, respectively. The principal wave numbers obtained in infrared spectrum and
their corresponding assignment (bond, compound type and functional group) were
characteristic for meloxicam.
Wave number
(cm-1)
KBr disk
1620
Wave number
(cm-1)
ATR
Assignment of wave numbers
1549.03, 1616.69,
N-H bending - Amides (10)
644.39, 711.40,
760.55
3291.17
3281.43
O-H stretching - Phenols
1301.41, 1346.38,
1343.41, 1446.92,
Ring stretching - Heteroaromatics
1457.05, 1530.28,
1549.03
1550.52
1185.26, 1346.38
1343.41
C-O stretching for phenols
1043.78, 1064.56,
1042.82, 1151.12,
C-H in plane bending aromatic
118.65, 1130.91,
1262.47
1153.05, 1161.82,
1185.26
713.54, 761.70,
711.40, 760.55
784.78, 798.30
1457.05
C-H out of plane bending-mdisubstituted
1446.92
C=C stretching aromatic
20
4000
3500
3000
2500
Fig. 19. Infrared spectrum of meloxicam (ATR)
Wavenumbers (cm-1)
2000
1500
1000
454.73
564.75
607.41
644.39
1000
526.69
10
940.03
798.30
845.03
855.81
1130.91
1153.05
1161.82
784.78
729.95
1043.78
677.66
761.70
940.82
713.54
826.86
1064.56
1217.57
1620.66
60
711.40
30
1118.65
1265.21
1185.26
1301.41
1346.38
65
760.55
40
1500
1042.82
2000
1151.12
2500
1457.05
50
1262.47
60
3000
1530.28
40
1343.41
3500
1616.69
35
1550.52
45
3291.17
%T
55
823.04
1446.92
70
3281.43
4000
1549.03
%T
78
75
70
30
25
20
Wavenumbers (cm-1)
500
Fig. 18. Infrared spectrum of meloxicam (KBr disk)
100
90
80
50
0
500
79
4.2.7 Infrared spectrum of celecoxib
The infrared spectrum of celecoxib in KBr disk and ATR is shown in Fig. 20 and
Fig. 21, respectively. The principal wave numbers obtained in infrared spectrum and their
corresponding assignment (bond, compound type and functional group) were
characteristic for celecoxib.
Wave number
(cm-1)
KBr disk
Wave number
(cm-1)
ATR
Assignment of wave numbers
1164.84 &
1347.72
1346
S=O stretching- Sulfonamide
3341.93
3332.02
N-H stretching
1135.35
742.11, 760.62,
791.14, 801.09,
844.77, 904.89,
969.40, 980.86,
1101.68, 1015.93,
1132.63, 1157.53,
1228.50, 1274.23,
1346.02, 1374.13
C-X stretching (X- Fluoride)
1473.65, 149.58,
1605.98
1473.94, 1497.15,
1562.01, 1593.78
C=N stretching
1016.33, 1093.86,
1135.35, 1164.84,
1229.98, 1274.78
1101.68, 1015.93,
1132.63, 1157.53,
1228.50, 1274.23
C-H in plane bending aromatic
761.88, 792.33,
801.78, 846.22
721.03, 760.62,
742.11, 791.14,
801.09, 844.77
C-H out of plane bending-p-
1402.51, 1473.94,
1445.84, 1497.15,
1593.78
C=C stretching aromatic
1446.64, 1473.65,
1498.58
substituted
4000
3500
3000
2500
Fig. 21. Infrared spectrum of celecoxib (ATR)
15
10
Wavenumbers (cm-1)
2000
20
1500
1000
742.11
25
647.60
499.54
630.77
441.66
904.89
421.67
721.03
1000
511.32
530.35
559.92
969.40
1015.93
1135.35
1164.84
846.22
1093.86
1274.78
801.78
1016.33
1229.98
1473.65
792.33
981.63
1446.64
1695.42
761.88
907.24
1605.98
65
760.62
35
1101.68
1500
791.14
40
1347.72
57
801.09
50
1228.50
53
1374.31
54
980.86
2000
1445.84
70
2500
1473.94
55
3000
1157.53
55
1498.58
56
1132.63
75
1497.15
3500
1561.01
58
844.77
30
1274.23
60
3235.17
64
1346.02
45
1593.78
59
3096.22
60
3341.93
62
1374.13
65
3332.02
%T
61
3226.12
4000
1402.51
%T
80
67
66
63
52
Wavenumbers (cm-1)
500
Fig. 20. Infrared spectrum of celecoxib (KBr disk)
85
80
5
-5
0
500
81
4.3 TLC analysis of NSAIDs
One of the most effective screening methods is the thin-layer chromatography
(TLC), simplest of all the widely used chromatographic methods to perform. The TLC
procedure was included to test the purity and for characterization of NSAIDs. Unlike in a
conventional method, Camag HPTLC system offers the advantages of automatic
application under the pressure of nitrogen gas and scanning in situ, where the conditions
can be more easily controlled. Beside, several samples can be run simultaneously using a
small quantity of mobile phase and the substances are permanently stored on the plate.
The mobile phase consisting of methanol: toluene (1:1 v/v) for aspirin and isopropyl
alcohol: n-hexane (4.9: 5.1 v/v) for other NSAIDs gave good resolution and sharp peaks
(Fig. 22 and 23). Also, the spots were compact and not diffused (Plate 1). It was observed
that pre-washing of TLC plates with methanol followed by drying and pre-saturation of
TLC chamber with mobile phase for 10 min ensured good reproducibility for peak shapes
of drugs. The chromatogramn in Fig. 22 and 23 outlines a single prominent peak in each
track represented by NSAIDs which is indicative of presence of drug or absence of
impurities. The densitogram obtained for NSAIDs is shown in the Plate 2.
The uv-absorbance spectra for NSAIDs diclofenac, paracetamol, ketoprofen,
celecoxib, nimesulide, meloxicam and aspirin are shown in Fig. 24, 25, 26, 27, 28, 29,
and 30, respectively. The maximum absorption wavelength for diclofenac, paracetamol,
ketoprofen, celecoxib, nimesulide, meloxicam and aspirin was recorded at 281, 246, 258,
307, 361 and 225, respectively.
82
Fig. 22: TLC chromatogram of NSAIDs
Fig. 23: TLC chromatogram of aspirin
83
Plate 1: Spotted plate post
Plate 2: Densitogram of meloxicam, ketoprofen, diclofenac, paracetamol, nimesulide
development under TLC visualizer.
and celecoxib
100
281
Absorption Unit
Absorption Unit
100
75
50
25
0
200
250
300
350
75
50
25
0
200
400
Absorption Unit
Absorption Unit
100
75
50
25
300
350
350
400
Fig 25: HPTLC-UV absorption spectra of paracetamol
263
250
300
84
Fig 24:HPTLC-UV absorption spectra of diclofenac
0
200
250
Wavelength in nm
Wavelength in nm
100
246
400
Wavelength in nm
Fig 26: HPTLC-UV absorption spectra of ketoprofen
258
75
50
25
0
200
250
300
350
400
Wavelength in nm
Fig 27: HPTLC-UV absorption spectra of celecoxib
307
75
50
25
0
200
250
300
350
400
450
500
Absorption Unit
100
75
225
25
250
300
350
Wavelength in nm
Fig 30: HPTLC-UV absorption spectra of aspirin
50
25
0
200
250
300
350
400
450
Wavelength in nm
Fig 28: HPTLC-UV absorption spectra of nimesulide
0
200
75
400
Fig 29: HPTLC-UV absorption spectra of meloxicam
85
Wavelength in nm
50
361
100
Absorption Unit
Absorption Unit
100
86
4.4 Clinical signs of toxicity
In the present study, the diclofenac received birds showed clinical manifestation
such as anorexia, dullness, ruffled feathers, disinclination to move within the cage,
lethargy, depression, recumbence (Plate 3), shrunken eyes and occult blood in the faecal
droppings. All these clinical signs were observed on day 2 onwards and was continued to
be so until the end of experiment.
The birds which had received aspirin and paracetamol showed clinical signs of
toxicity such as dullness, ruffled feathers and disinclination to move. These clinical signs
were observed 2 to 3 h after daily dosing. But, over a period of time (6-7 h after dosing),
the observed clinical signs subsided indicating the observed clinical signs of toxicity were
transitory. In addition, these birds showed watery droppings accompanied with blood
mixed mucous. On the contrary to the diclofenac group, aspirin and paracetamol received
birds did not show signs of anorexia and in fact these birds consumed feed very well.
The birds which received ketoprofen, nimesulide, meloxicam and celecoxib
showed dullness and occasionally mucous mixed watery droppings otherwise these birds
did not show any clinical signs of toxicity.
4.5 Mortality
The diclofenac (Group II) received birds showed mortality on day 3, 4 and 5. Four
out of 6 birds died during the experimental period, whereas, no mortality was observed in
the birds, which received aspirin, paracetamol, nimesulide, ketoprofen, meloxicam and
celecoxib.
87
4.6 Hematology Parameters
4.6.1 RBC count (×106/l)
The RBC count (×106/l) in Group II birds administered with diclofenac (2.5
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 2.91±0.15, 2.70±0.20, 2.15±0.12, 2.04±0.14
1.84±0.12 and 1.68±0.21, respectively. Except for day 1 and 2, RBC count for all other
days (3, 4, 5 and 6) was significantly (P<0.01) lower compared to control group values of
2.80±0.10, 2.80±0.08, 2.78±0.09 and 2.74±0.10, respectively (Table 2 and Fig. 31).
The RBC count (×106/l) in Group III birds administered with aspirin (10 mg/kg,
PO) on days 1, 2, 3, 4, 5 and 6 was 2.91±0.15, 2.70±0.20, 2.15±0.12, 2.04±0.14,
1.84±0.12 and 1.68±0.21, respectively. The RBC count on day 4, 5 and 6 was
significantly (P<0.01) lower compared to control group value of 2.80±0.08, 2.78±0.09
and 2.74±0.10, respectively (Table 2 and Fig. 31).
The RBC count (×106/l) in Group V birds administered with nimesulide (2
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 2.94±0.05, 2.86±0.04, 2.74±0.15, 2.60±0.09,
2.25±0.09 and 2.20±0.20, respectively. The blood RBC count on day 5 and 6 was
significantly (P<0.05) lower compared to control group value of 2.78±0.09 and
2.74±0.10, respectively (Table 2 and Fig. 31).
Other treated groups did not show any significant (P>0.05) alterations in the
values of RBC count compared to the control group values.
88
Table 2: Effect of oral administration of NSAIDs on RBC count (106/l) in broiler chickens
RBC count (106/l)
Groups
Day
2
4
5
6
2.94±0.05
2.84±0.06
2.80±0.10
2.80±0.08
2.78±0.09
2.74±0.10
Group II
(Diclofenac)
2.91±0.15
2.70±0.20
2.15±0.12**
n=5
2.04±0.14**
n=3
1.84±0.12**
n=2
1.68±0.21***
n=2
Group III
(Aspirin)
2.93±0.15
2.80±0.12
2.78±0.12
2.20±0.19**
2.10±0.15**
2.00±0.12***
Group IV
(Paracetamol)
2.90±0.15
2.85±0.09
2.81±0.12
2.80±0.14
2.78±0.18
2.75±0.10
Group V
(Nimesulide)
2.94±0.05
2.86±0.04
2.74±0.15
2.60±0.09
2.25±0.09*
2.20±0.20*
Group VI
(Ketoprofen)
2.85±0.15
2.83±0.12
2.80±0.08
2.79±0.14
2.78±0.17
2.77±0.11
Group VII
(Meloxicam)
2.93±0.14
2.89±0.20
2.85±0.10
2.84±0.12
2.82±0.13
2.80±0.09
Group VIII
(Celecoxib)
2.96±0.12
2.84±0.13
2.84±0.10
2.82±0.10
2.81±0.09
2.79±0.15
Group I
(Control)
3
Values are Mean±SE; For each group n=6 unless otherwise mentioned
*P<0.05, *P<0.01, ***P<0.001 in relation to control.
88
1
89
3.0
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
2.0
6
RBC count (10 /l)
2.5
89
1.5
1.0
0.5
0.0
1
2
3
4
5
Day
Fig. 31: Effect of oral administration of NSAIDs on RBC count (106/l) in broiler chickens
6
90
4.6.2 Hemoglobin concentration (g/dl)
The hemoglobin concentration (g/dl) in Group II birds administered with
diclofenac (2.5 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 12.90±0.14, 12.60±0.15,
12.41±0.12, 11.85±0.09, 10.85±0.11 and 10.01±0.10, respectively. Except for day 1 and
2, hemoglobin concentration (g/dl) for all other days (3, 4, 5 and 6) was significantly
(P<0.01) lower compared to control group values of 12.89±0.08, 12.60±0.13, 12.56±0.12
and 12.50±0.11, respectively (Table 3 and Fig. 32).
The hemoglobin concentration (g/dl) in Group III birds administered with aspirin
(10 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 12.40±0.11, 12.10±0.12, 12.00±0.15 and
11.75±0.07, respectively. The hemoglobin concentration (g/dl) on days 3, 4, 5 and 6 was
significantly (P<0.05) lower compared to control group value of 12.89±0.08, 12.60±0.13,
12.56±0.12 and 12.50±0.11, respectively (Table 3 and Fig. 32).
The hemoglobin concentration (g/dl) in Group V birds administered with
nimesulide (3 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 13.12±0.14, 12.98±0.12,
12.45±0.13, 12.12±0.11, 12.07±0.10 and 11.90±0.07, respectively. The hemoglobin
concentration (g/dl) on days 4, 5 and 6 was significantly (P<0.05) lower compared to
control group value of 12.60±0.13, 12.56±0.12 and 12.50±0.11, respectively (Table 3 and
Fig. 32).
Other treated groups did not show any significant (P>0.05) alterations in
hemoglobin concentration (g/dl) compared to the control group values.
Table 3: Effect of oral administration of NSAIDs on hemoglobin concentration (g/dl) in broiler chickens
Hemoglobin concentration (g/dl)
Groups
Day
1
Group I
(Control)
2
3
4
5
6
13.00±0.09
12.89±0.08
12.60±0.13
12.56±0.12
12.50±0.11
Group II
(Diclofenac)
12.90±0.14
12.60±0.15
12.41±0.12*
n=5
11.85±0.09**
n=3
10.85±0.11***
n=2
10.01±0.10***
n=2
Group III
(Aspirin)
13.10±0.10
12.80±0.12
12.40±0.11*
12.10±0.12*
12.00±0.15**
11.75±0.07***
Group IV
(Paracetamol)
13.20±0.12
13.00±0.13
12.90±0.14
12.80±0.12
12.80±0.12
12.75±0.15
Group V
(Nimesulide)
13.12±0.14
12.98±0.12
12.45±0.13
12.12±0.11*
12.07±0.10*
11.90±0.07**
Group VI
(Ketoprofen)
13.01±0.11
13.20±0.10
12.90±0.09
12.91±0.12
12.70±0.11
12.46±0.10
Group VII
(Meloxicam)
12.98±0.10
12.90±0.11
12.84±0.14
12.71±0.08
12.74±0.07
12.54±0.14
Group VIII
(Celecoxib)
12.84±0.12
12.86±0.14
12.74±0.13
12.78±0.15
12.56±0.14
12.61±0.12
Values are Mean±SE; For each group n=6 unless otherwise mentioned
P<0.05, **P<0.01, ***P<0.001 in relation to control.
91
13.01±0.12
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
13
11
10
9
92
Hemoglobin concentration (g/dl)
12
8
7
6
5
4
3
2
1
0
1
2
3
4
5
6
Day
Fig. 32: Effect of oral administration of NSAIDs on hemoglobin concentration (g/dl) in broiler chickens
93
4.6.3 Packed Cell Volume (PCV) (%)
The PCV (%) in Group II birds administered with diclofenac (2.5 mg/kg, PO) on
days 1, 2, 3, 4, 5 and 6 was 33.47±0.12, 32.41±0.21, 31.98±0.13, 31.50±0.25, 30.54±0.26
and 29.41±0.10, respectively. Except for day 1 and 2, PCV (%) for all other days (3, 4, 5
and 6) was significantly (P<0.01) lower compared to control group values of 32.80±0.30,
32.71±0.14, 32.00±0.31 and 31.53±0.21, respectively (Table 4 and Fig. 33).
The PCV (%) in Group III birds administered with aspirin (10 mg/kg, PO) on
days 1, 2, 3, 4, 5 and 6 was 33.26±0.21, 33.10±0.23, 31.98±0.10, 31.80±0.13, 31.14±0.14
and 30.48±0.15, respectively. The PCV (%) on day 3, 4, 5 and 6 was significantly
(P<0.05) lower compared to control group value of 32.80±0.30, 32.71±0.14, 32.00±0.31
and 31.53±0.21, respectively (Table 4 and Fig. 33).
The PCV (%) in Group V birds administered with nimesulide (2 mg/kg, PO) on
days 1, 2, 3, 4, 5 and 6 was 33.02±0.14, 33.10±0.13, 32.87±0.21, 32.67±0.31, 31.10±0.21
and 30.44±0.17, respectively. The PCV (%) on day 4, 5 and 6 was significantly (P<0.05)
lower compared to control group value of 32.71±0.14, 32.00±0.31 and 31.53±0.21,
respectively (Table 4 and Fig. 33).
Other treated groups did not show any significant (P>0.05) alterations in the
values of PCV (%) compared to the control group values.
4.6.4 Platelet count (×103/l)
The platelet count (×103/l) in Group II birds administered with diclofenac (2.5
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 20±1.21, 16±1.30, 14±1.11, 12±2.06, 11±1.40
and 9±1.69, respectively. Except for day 1, platelet count for all other days (2, 3, 4, 5 and
94
6) was significantly (P<0.01) lower compared to control group values of 22±1.30,
20±2.01, 20±1.65, 21±1.42 and 19±1.84, respectively (Table 5 and Fig. 34).
The platelet count (×103/l) in Group III birds administered with aspirin (10
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 22±1.32, 18±1.47, 14±1.60, 13±1.65, 9±1.70
and 7±2.08, respectively. The platelet count on day 3, 4, 5 and 6 was significantly
(P<0.05) lower compared to control group value of 20±2.01, 20±1.65, 21±1.42 and
19±1.84, respectively (Table 5 and Fig. 34).
The platelet count (×103/l) in Group IV birds administered with paracetamol (10
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 22±1.04, 20±1.54, 19±1.33, 17±1.27, 14±1.52
and 12±1.12, respectively. The platelet count on day 5 and 6 was significantly (P<0.01)
lower compared to control group value of 21±1.42 and 19±1.84, respectively (Table 5
and Fig. 34).
The platelet count (×103/l) in Group V birds administered with nimesulide (2
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 24±1.22, 19±1.14, 20±1.26, 16±1.33, 16±1.21
and 13±1.49, respectively. The platelet count on day 6 was significantly (P<0.05) lower
compared to control group value of 19±1.84, respectively (Table 5 and Fig. 34).
The platelet count (×103/l) in Group VI birds administered with ketoprofen (4
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 23±1.01, 21±2.07, 17±2.14, 14±1.10, 13±1.30
and11±1.10, respectively. The platelet count on day 4, 5 and 6 was significantly
(P<0.001) lower compared to control group value of 20±1.65, 21±1.42 and 19±1.84,
respectively (Table 5 and Fig. 34).
The platelet count (×103/l) in Group VII birds administered with meloxicam (0.5
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 22±1.21, 21±1.32, 16±1.41, 14±1.28, 12±1.33
95
and 11±1.43, respectively. The platelet count on day 4, 5 and 6 was significantly
(P<0.001) lower compared to control group value of 20±1.65, 21±1.42 and 19±1.84,
respectively (Table 5 and Fig. 34).
The platelet count (×103/l) in Group VIII birds administered with celecoxib (3.5
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 20±1.24, 21±1.56, 18±1.31, 17±1.47, 19±1.40
and 13±1.61, respectively. The platelet count on day 6 was significantly (P<0.05) lower
compared to control group value of 19±1.84, respectively (Table 5 and Fig. 34).
Other treated groups did not show any significant (P>0.05) alterations in the
values of PCV (%) compared to the control group values.
4.7 Serum biochemical parameters
Serum obtained from blood samples collected on day 1 through day 6 of
experimental period were used for estimation of aspartate aminotransferase (AST),
alanine aminotransferase (ALT), alkaline phosphatase (ALP), blood urea nitrogen
(BUN), creatinine, uric acid, albumin and total protein. The obtained results for the above
said parameters are presented in Table 6-13 and Fig. 35-43.
4.7.1 Aspartate aminotransferase (AST) concentration
The AST concentration (U/L) in Group II birds administered with diclofenac (2. 5
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 71.87±5.79, 90.56±3.40, 140.93±5.02,
193.30±6.53, 212.09±3.46 and 226.28±13.83, respectively. Except for day 1 and 2, serum
AST concentration for all other days (3, 4, 5 and 6) was significantly (P<0.001) higher
compared to control group values of 97.80±0.71, 98.67±0.54, 97.72±2.58 and
99.84±2.96, respectively (Table 6 and Fig. 35).
Table 4: Effect of oral administration of NSAIDs on PCV (%) in broiler chickens
PCV (%)
Groups
Day
1
Group I
(Control)
2
3
4
5
6
33.00±0.23
32.80±0.30
32.71±0.14
32.00±0.31
31.53±0.21
Group II
(Diclofenac)
33.47±0.12
32.41±0.21
31.98±0.13*
n=5
31.50±0.25**
n=3
30.54±0.26**
n=2
29.41±0.10***
n=2
Group III
(Aspirin)
33.26±0.21
33.10±0.23
31.98±0.10*
31.80±0.13*
31.14±0.14*
30.48±0.15**
Group IV
(Paracetamol)
33.54±0.14
33.05±0.15
32.84±0.21
32.78±0.23
32.07±0.21
31.24±0.13
Group V
(Nimesulide)
33.02±0.14
33.10±0.13
32.87±0.21
32.67±0.31
31.10±0.21*
30.44±0.17**
Group VI
(Ketoprofen)
33.10±0.14
33.00±0.15
32.79±0.14
32.56±0.20
32.45±0.27
32.15±0.29
Group VII
(Meloxicam)
33.56±0.21
33.21±0.23
33.41±0.21
33.10±0.25
32.19±0.24
32.14±0.26
Group VIII
(Celecoxib)
33.14±0.21
33.52±0.23
33.04±0.24
32.98±0.21
32.40±0.23
32.25±0.18
Values are Mean±SE; For each group n=6 unless otherwise mentioned
*P<0.05, **P<0.01, ***P<0.001 in relation to control.
96
33.12±0.21
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
30
25
PCV (%)
97
20
15
10
5
0
1
2
3
4
5
Day
Fig. 33 : Effect of oral administration of NSAIDs on PCV (%) in broiler chickens
6
Table 5: Effect of oral administration of NSAIDs on platelet count (103/l) in broiler chickens
Platelet count (103/l)
Groups
Day
1
Group I
(Control)
2
3
4
5
6
22±1.30
20±2.01
20±1.65
21±1.42
19±1.84
Group II
(Diclofenac)
20±1.21
16±1.30*
14±1.11*
n=5
12±2.06**
n=3
11±1.40**
n=2
9±1.69**
n=2
Group III
(Aspirin)
22±1.32
18±1.47
14±1.60*
13±1.65**
9±1.70***
7±2.08***
Group IV
(Paracetamol)
22±1.04
20±1.54
19±1.33
17±1.27
14±1.52**
12±1.12**
Group V
(Nimesulide)
24±1.22
19±1.14
20±1.26
16±1.33
16±1.21
13±1.49*
Group VI
(Ketoprofen)
23±1.01
21±2.07
17±2.14
14±1.10*
13±1.30***
11±1.10***
Group VII
(Meloxicam)
22±1.21
21±1.32
16±1.41
14±1.28*
12±1.33***
11±1.43***
Group VIII
(Celecoxib)
20±1.24
21±1.56
18±1.31
17±1.47
19±1.40
13±1.61*
Values are Mean±SE; For each group n=6 unless otherwise mentioned
*P<0.05, P<0.01, ***P<0.001 in relation to control.
98
21±1.23
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
25
15
99
Platelet count (103/l)
20
10
5
0
1
2
3
4
5
Day
Fig. 34: Effect of oral administration of NSAIDs on platelet count (103/l) in broiler chickens
6
100
The AST concentration (U/L) in Group V birds administered with nimesulide (2
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 80.01±4.17, 87.69±2.56, 97.96±0.59,
100.34±1.02, 107.78±1.58 and 115.16±1.75, respectively. The serum AST concentration
on day 6 was significantly (P<0.01) higher compared to control group I value of
99.84±2.96, respectively (Table 6 and Fig. 35).
Other treated groups did not show any significant (P>0.05) alterations in the
values of AST compared to the control group values.
4.7.2 Alanine aminotransferase (ALT) concentration
The ALT concentration (U/L) in Group II birds administered with diclofenac (2. 5
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 16.21±0.99, 41.96±3.47, 53.18±3.32,
60.54±2.72, 64.43±2.14 and 70.16±1.95, respectively. Except for day 1, serum ALT
concentration for all other days (2, 3, 4, 5 and 6) was significantly (P<0.001) higher
compared to control group values of 19.25±1.39, 19.87±1.04, 20.78±0.53, 21.17±0.52
and 20.72±0.41, respectively (Table 7 and Fig. 36).
The ALT concentration (U/L) in Group V birds administered with nimesulide (2
mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 17.09±1.29, 19.25±1.28, 21.99±0.97,
21.90±0.84, 26.37±0.62 and 30.04±0.30, respectively. The serum ALT concentration on
day 5 and 6 was significantly (P< 0.05) higher compared to control group value of
21.17±0.52 and 20.72±0.41, respectively (Table 7 and Fig. 36).
Other treated groups did not show any significant (P>0.05) alterations in the
values of ALT compared to the control group values.
Table 6: Effect of oral administration of NSAIDs on aspartate aminotransferase (AST) concentration in broiler chickens
Asparatae aminotransferase (U/L)
Groups
Day
1
Group I
(Control)
2
3
4
5
6
82.69±5.87
97.80±0.71
98.67±0.54
97.72±2.58
99.84±2.96
Group II
(Diclofenac)
71.87±5.79
90.56±3.40
140.93±5.02***
n=5
193.30±6.53***
n=3
212.09±3.46***
n=2
226.28±13.83***
n=2
Group III
(Aspirin)
75.85±2.91
80.11±2.51
87.68±2.60
103.64±2.28
109.90±2.27
112.41±1.21
Group IV
(Paracetamol)
69.75±4.04
79.75±6.74
91.52±5.55
103.14±2.09
108.73±1.90
110.41±2.35
Group V
(Nimesulide)
80.01±4.17
87.69±2.56
97.96±0.59
100.34±1.02
107.78±1.58
115.16±1.75**
Group VI
(Ketoprofen)
74.66±4.19
82.12±4.35
90.29±3.74
95.58±1.24
102.26±1.88
109.66±1.07
Group VII
(Meloxicam)
73.00±4.33
76.95±3.02
87.46±2.97
97.25±1.53
100.76±1.14
110.33±2.20
Group VIII
(Celecoxib)
74.66±2.64
78.95±4.59
88.63±3.54
99.25±2.07
97.26±3.70
108.83±4.93
Values are Mean±SE; For each group n=6 unless otherwise mentioned
**P<0.01, ***P<0.001 in relation to control.
101
78.35±4.83
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
250
225
175
150
102
AST concentration (U/L)
200
125
100
75
50
25
0
1
2
3
4
5
6
Day
Fig. 35: Effect of oral administration of NSAIDs on serum AST concentration (U/L) in broiler chickens
Table 7: Effect of oral administration of NSAIDs on alanine aminotransferase (ALT) concentration in broiler chickens
Alanine aminotransferase (U/L)
Groups
Day
1
Group I
(Control)
2
3
4
5
6
19.25±1.39
19.87±1.04
20.78±0.53
21.17±0.52
20.72±0.41
Group II
(Diclofenac)
16.21±0.99
41.96±3.47***
53.18±3.32***
n=5
60.54±2.72***
n=3
64.43±2.14***
n=2
70.16±1.95***
n=2
Group III
(Aspirin)
18.55±1.54
19.76±2.57
19.77±0.37
21.39±0.83
21.70±0.43
22.23±0.47
Group IV
(Paracetamol)
18.76±2.89
18.58±1.32
20.34±1.15
21.90±0.84
21.99±0.97
25.48±3.06
Group V
(Nimesulide)
17.09±1.29
19.25±1.28
21.99±0.97
21.90±0.84
26.37±0.62
30.04±0.30**
Group VI
(Ketoprofen)
20.46±0.49
20.92±0.71
21.20±1.35
21.11±0.56
21.00±0.56
20.72±0.41
Group VII
(Meloxicam)
19.42±0.29
20.54±1.39
19.10±1.04
21.63±0.53
20.17±0.82
21.70±0.43
Group VIII
(Celecoxib)
20.13±0.21
21.08±0.74
22.20±0.98
21.44±0.30
20.17±0.82
20.89±0.48
Values are Mean±SE; For each group n=6 unless otherwise mentioned
**P<0.01, ***P<0.001 in relation to control.
103
19.63±0.29
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
50
104
ALT concentration (U/L)
75
25
0
1
2
3
4
5
6
Day
Fig. 36 : Effect of oral administration of NSAIDs on serum ALT concentration (U/L) in broiler chickens
105
4.7.3 Alkaline phosphatase (ALP) concentration
The ALP concentration (KA units/dl) in Group II birds administered with
diclofenac (2.5 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 15.16±0.47, 18.83±0.30,
24.00±0.70, 26.66±0.66, 32.500±0.50 and 35.50±0.50, respectively. Except for day 1,
serum ALP concentration for all other days (2, 3, 4, 5 and 6) was significantly (P<0. 001)
higher compared to control group values of 15.43±0.42, 15.33±0.42, 16.16±0.79,
17.66±0.33 and 17.33±0.61, respectively (Table 8 and Fig. 37).
The ALP concentration (KA units/dl) in Group IV birds administered with
paracetamol (10. mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 16.16±0.47, 16.00±0.68,
16.16±0.47, 18.00±0.36, 19.66±0.49 and 20.00±0.93, respectively. The serum ALP
concentration on day 6 was significantly (P<0.01) higher compared to control group
value of 17.33±0.61 (Table 8 and Fig. 37).
The ALP concentration (KA units/dl) in Group V birds administered with
nimesulide (2 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 16.50±0.76, 17.00±0.63,
15.66±0.49, 17.83±0.30, 20.16±0.60 and 22.66±0.42, respectively. The serum ALP
concentration on day 5 and 6 was significantly (P<0.01) higher compared to control
group value of 17.66±0.33 and 17.33±0.61, respectively (Table 8 and Fig. 37).
The ALP concentration (KA units/dl) in Group VI birds administered with
ketoprofen (4 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 15.66±0.66, 15.50±0.42,
16.33±0.66, 17.83±0.30, 18.00±0.25 and 19.66±0.71, respectively. The serum ALP
concentration on day 6 was significantly (P<0.05) higher compared to control group
value of 17.33±0.61 (Table 8 and Fig. 37).
Table 8: Effect of oral administration of NSAIDs on alkaline phosphatase (ALP) concentration in broiler chickens
Alkaline phosphatase (KA units/dl)
Groups
Day
1
Group I
(Control)
2
3
4
5
6
15.43±0.42
15.33±0.42
16.16±0.79
17.66±0.33
17.33±0.61
Group II
(Diclofenac)
15.16±0.47
18.83±0.30***
24.00±0.70***
n=5
26.66±0.66***
n=3
32.50±0.50***
n=2
35.50±0.50***
n=2
Group III
(Aspirin)
15.16±0.47
15.33±0.49
14.83±0.30
18.16±0.477
17.33±0.33
16.83±0.70
Group IV
(Paracetamol)
16.16±0.47
16.00±0.68
16.16±0.47
18.00±0.36
19.66±0.49
20.00±0.93**
Group V
(Nimesulide)
16.50±0.76
17.00±0.63
15.66±0.49
17.83±0.30
20.16±0.60**
22.66±0.42***
Group VI
(Ketoprofen)
15.66±0.66
15.50±0.42
16.33±0.66
17.83±0.30
18.00±0.25
19.66±0.71*
Group VII
(Meloxicam)
15.33±0.55
15.33±0.33
17.00±0.51
16.66±0.71
18.33±0.55
19.16±0.54
Group VIII
(Celecoxib)
15.00±0.36
16.16±0.47
15.83±0.60
16.83±0.47
16.33±0.49
17.00±0.68
Values are Mean±SE; For each group n=6 unless otherwise mentioned
*P<0.05, **P<0.01, ***P<0.001 in relation to control.
106
15.00±0.51
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
107
ALP concentration (KA units/dl)
30
20
10
0
1
2
3
4
5
6
Day
Fig. 37: Effect of oral administration of NSAIDs on serum ALP concentration (KA units/dl) in broiler chickens
108
Other treated groups did not show any significant (P>0.05) alterations in the
values of ALP concentration compared to the control group values.
4.7.4 Blood Urea Nitrogen concentration (BUN)
The blood urea nitrogen concentration (mg/dl) in Group II birds administered
with diclofenac (2.5 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 3.73±0.06, 4.75±0.06,
5.18±0.08, 5.46±0.17, 6.60±0.20 and 7.25±0.25, respectively. Except for day 1, BUN
concentration for all other days (2, 3, 4, 5 and 6) was significantly (P<0.001) higher
compared to control group values of 3.51±0.12, 3.78±0.13, 3.85±0.20, 3.95±0.20 and
4.75±0.08, respectively (Table 9 and Fig. 38).
Other treated groups did not show any significant (P>0.05) alterations in the
values of creatinine compared to the control group values.
4.7.5 Creatinine concentration (mg/dl)
The creatinine concentration (mg/dl) in Group II birds administered with
diclofenac (2.5 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 0.35±0.03, 0.48±0.04,
0.65±0.01, 0.77±0.01, 0.83±0.03 and 0.89±0.00, respectively. Except for day 1 and 2,
serum creatinine concentration for all other days (3, 4, 5 and 6) was significantly (P<0.
001) higher compared to control group values of 0.37±0.03, 0.42±0.02, 0.42±0.03,
0.42±0.03, respectively (Table 10 and Fig. 39).
The creatinine concentration (mg/dl) in Group V birds administered with
nimesulide (2 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 0.39±0.02, 0.40±0.02,
0.35±0.00, 0.43±0.02, 0.47±0.00 and 0.54±0.01, respectively. The serum creatinine
concentration on day 6 was significantly (P< 0. 01) higher compared to control group
value of 0.42±0.03 (Table 10 and Fig. 39).
Table 9: Effect of oral administration of NSAIDs on BUN concentration (mg/dl) in broiler chickens
Blood Urea Nitrogen (mg/dl)
Groups
Day
1
Group I
(Control)
2
3
4
5
6
3.51±0.12
3.78±0.13
3.85±0.20
3.95±0.20
4.75±0.08
Group II
(Diclofenac)
3.73±0.06
4.75±0.06***
5.18±0.08***
n=5
5.46±0.17***
n=3
6.60±0.20***
n=2
7.25±0.25***
n=2
Group III
(Aspirin)
3.76±0.06
3.85±0.04
3.83±0.08
3.85±0.04
3.95±0.07
4.48±0.04
Group IV
(Paracetamol)
3.43±0.05
3.68±0.06
3.86±0.08
4.00±0.05
3.88±0.04
4.55±0.08
Group V
(Nimesulide)
3.75±0.07
3.73±0.06
3.68±0.11
3.88±0.06
3.91±0.07
4.45±0.11
Group VI
(Ketoprofen)
3.86±0.04
3.70±0.05
4.00±0.06
4.11±0.07
4.01±0.09
4.45±0.12
Group VII
(Meloxicam)
3.90±0.02
3.76±0.03
4.08±0.09
4.10±0.05
4.03±0.06
4.71±0.07
Group VIII
(Celecoxib)
3.81±0.09
3.85±0.04
4.06±0.04
4.13±0.05
4.08±0.06
4.61±0.08
Values are Mean±SE; For each group n=6 unless otherwise mentioned
***P<0.001 in relation to control.
109
3.66±0.16
Control
Diclofenac
Aspirin
Pracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
8
7
5
110
BUN concentration (mg/dl)
6
4
3
2
1
0
1
2
3
4
5
Day
Fig. 38: Effect of oral administration of NSAIDs on BUN concentration (mg/dl) in broiler chicken
6
Table 10: Effect of oral administration of NSAIDs on serum creatinine concentration (mg/dl) in broiler chickens
Creatinine (mg/dl)
Groups
Day
1
Group I
(Control)
2
3
4
5
6
0.43±0.03
0.37±0.03
0.42±0.02
0.42±0.03
0.42±0.03
Group II
(Diclofenac)
0.35±0.03
0.48±0.04
0.65±0.01***
n=5
0.77±0.01***
n=3
0.83±0.03***
n=2
0.89±0.00***
n=2
Group III
(Aspirin)
0.42±0.04
0.38±0.02
0.37±0.01
0.38±0.02
0.38±0.01
0.40±0.02
Group IV
(Paracetamol)
0.40±0.03
0.35±0.01
0.38±0.01
0.44±0.03
0.41±0.03
0.46±0.03
Group V
(Nimesulide)
0.39±0.02
0.40±0.02
0.35±0.00
0.43±0.02
0.47±0.00
0.545±0.01**
Group VI
(Ketoprofen)
0.37±0.02
0.35±0.01
0.38±0.01
0.44±0.02
0.45±0.03
0.50±0.01
Group VII
(Meloxicam)
0.36±0.02
0.37±0.01
0.36±0.01
0.35±0.00
0.36±0.00
0.39±0.01
Group VIII
(Celecoxib)
0.38±0.02
0.34±0.01
0.38±0.01
0.44±0.03
0.40±0.02
0.41±0.02
Values are Mean+SE ; For each group n=6 unless otherwise mentioned
**P<0.01, ***P<0.001 in relation to control.
111
0.38±0.02
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
0.9
0.7
0.6
112
Creatinine concentration (mg/dl)
0.8
0.5
0.4
0.3
0.2
0.1
0.0
1
2
3
4
5
6
Day
Fig. 39: Effect of oral administration of NSAIDs on serum creatinine concentration (mg/dl) in broiler chickens
113
4.7.6 Uric acid concentration (mg/dl)
The uric acid concentration (mg/dl) in Group II birds administered with
diclofenac (2.5 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 3.95±0.07, 4.20±0.02,
6.36±0.15, 7.51±0.05, 8.68±0.13 and 9.45±0.04, respectively. Except for day 1, serum
uric acid concentration for all other days (2, 3, 4, 5 and 6) was significantly (P<0. 05)
higher compared to control group values of 3.80±0.13, 4.31±0.06, 4.13±0.08, 4.66±0.14
and 4.90±0.03, respectively (Table 11 and Fig. 40).
The uric acid concentration (mg/dl) in Group VI birds administered with
ketoprofen (4 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 3.78±0.09, 3.75±0.07,
4.23±0.06, 4.16±0.10, 4.51±0.04 and 5.31±0.14, respectively. The serum uric acid
concentration on day 6 was significantly (P<0.05) higher compared to control group
value of 4.90±0.03, respectively (Table 11 and Fig. 40).
Other treated groups did not show any significant (P>0.05) alterations in the uric
acid concentration compared to the control group values.
4.7.7 Serum albumin concentration (g/dl)
The serum albumin concentration (g/dl) in Group II birds administered with
diclofenac (2.5 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 1.85±0.04, 1.80±0.04,
1.64±0.02, 1.53±0.03, 1.60±0.10 and 1.50±0.10, respectively. Except for day 1 and 2,
serum albumin concentration for all other days (3, 4, 5 and 6) was significantly (P<0.01)
higher compared to control group values of 1.95±0.07, 1.96±0.08, 2.10±0.10 and
2.13±0.12, respectively (Table 12 and Fig. 41).
The serum albumin concentration (g/dl) in Group III birds administered with
aspirin (10 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 1.56±0.06, 2.20±0.06, 2.16±0.06,
114
2.05±0.07, 1.83±0.02 and 1.76±0.04, respectively. The serum albumin concentration on
day 5 and 6 was significantly (P<0.05) higher compared to control group value of
2.10±0.10 and 2.13±0.12, respectively (Table 12 and Fig. 41).
Other treated groups did not show any significant (P>0.05) alterations in the
serum albumin concentration compared to the control group values.
4.7.8 Serum Total protein (g/dl)
The total protein concentration (g/dl) in Group II birds administered with
diclofenac (2. 5 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 4.95±0.09, 4.56±0.24,
4.38±0.09, 4.20±0.06, 3.90±0.10 and 3.85±0.00, respectively. Except for day 1, serum
albumin concentration for all other days (2, 3, 4, 5 and 6) was significantly (P<0.01)
lower compared to control group values of 5.21±0.14, 5.03±0.27, 5.18±0.18, 5.96±0.06
and 5.85±0.10, respectively (Table 13 and Fig. 42).
The total protein concentration (g/dl) in Group III birds administered with aspirin
(10 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 4.66±0.06, 5.18±0.15, 5.08±0.11,
5.21±0.21, 5.18±0.06 and 5.01±0.16, respectively. The serum total protein concentration
on day 5 and 6 was significantly (P<0.001) lower compared to control group value of
5.96±0.06 and 5.85±0.10, respectively (Table 13 and Fig. 42).
The total protein concentration (g/dl) in Group IV birds administered with
paracetamol (10 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 4.75±0.15, 4.86±0.12,
5.13±0.15, 5.11±0.06, 5.30±0.03 and 5.08±0.15, respectively. The serum total protein
concentration on day 5 and 6 was significantly (P< 0. 01) lower compared to control
group value of 5.96±0.06 and 5.85±0.10, respectively (Table 13 and Fig. 42).
Table 11: Effect of oral administration of NSAIDs on serum uric acid concentration (mg/dl) in broiler chickens
Uric acid (mg/dl)
Groups
Day
1
Group I
(Control)
Group II
(Diclofenac)
2
3
4
5
6
3.80±0.13
4.31±0.06
4.13±0.08
4.66±0.14
4.90±0.03
3.95±0.07
4.20±0.02*
6.36±0.15***
n=5
7.51±0.05***
n=3
8.68±0.13***
n=2
9.45±0.04***
n=2
Group III
(Aspirin)
3.78±0.07
4.05±0.07
4.15±0.09
4.21±0.07
4.50±0.09
5.21±0.08
Group IV
(Paracetamol)
3.83±0.03
4.00±0.05
4.05±0.07
4.40±0.05
4.60±0.01
5.00±0.09
Group V
(Nimesulide)
3.93±0.11
4.08±0.06
4.01±0.07
4.20±0.03
4.70±0.05
5.25±0.07
Group VI
(Ketoprofen)
3.78±0.09
3.75±0.07
4.23±0.06
4.16±0.10
4.51±0.04
5.31±0.14*
Group VII
(Meloxicam)
3.98±0.06
3.95±0.09
4.00±0.07
4.40±0.20
4.78±0.11
5.23±0.04
Group VIII
(Celecoxib)
3.95±0.06
4.06±0.07
4.31±0.10
4.03±0.08
4.88±0.13
4.88±0.04
Values are Mean+SE ; For each group n=6 unless otherwise mentioned
*P<0.05, ***P<0.001 in relation to control.
115
4.10±0.15
10
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
9
7
6
5
116
Uric acid concentration (mg/dl)
8
4
3
2
1
0
1
2
3
4
5
6
Day
Fig. 40: Effect of oral administration of NSAIDs on serum uric acid concentration (mg/dl) in broiler chickens
Table 12: Effect of oral administration of NSAIDs on serum albumin concentration (g/dl) in broiler chickens
Albumin (g/dl)
Groups
Day
1
Group I
(Control)
2
3
4
5
6
1.96±0.16
1.95±0.07
1.96±0.08
2.10±0.10
2.13±0.12
Group II
(Diclofenac)
1.85±0.04
1.80±0.04
1.64±0.02*
n=5
1.53±0.03**
n=3
1.60±0.10**
n=2
1.50±0.10***
n=2
Group III
(Aspirin)
1.56±0.06
2.20±0.06
2.16±0.06
2.05±0.07
1.83±0.02*
1.76±0.04**
Group IV
(Paracetamol)
1.65±0.07
1.90±0.11
1.95±0.05
2.06±0.13
2.03±0.06
2.35±0.11
Group V
(Nimesulide)
2.11±0.06
2.15±0.10
2.13±0.04
2.08±0.06
2.20±0.06
2.23±0.08
Group VI
(Ketoprofen)
1.85±0.05
2.01±0.09
2.15±0.09
1.93±0.07
2.36±0.07
2.16±0.04
Group VII
(Meloxicam)
1.81±0.03
1.75±0.04
1.85±0.04
2.15±0.13
2.08±0.05
2.26±0.04
Group VIII
(Celecoxib)
2.08±0.03
2.11±0.06
1.83±0.02
1.85±0.04
2.15±0.05
2.15±0.04
Values are Mean+SE ; For each group n=6 unless otherwise mentioned
*P<0.05, **P<0.01, ***P<0.001 in relation to control.
117
1.85±0.06
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
2.0
1.5
118
Albumin concentration (g/dl)
2.5
1.0
0.5
0.0
1
2
3
4
5
6
Day
Fig. 41: Effect of oral administration of NSAIDs on serum albumin concentration (g/dl) in broiler chickens
119
The total protein concentration (g/dl) in Group V birds administered with
nimesulide (2 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 4.95±0.14, 5.00±0.15,
5.33±0.07, 5.16±0.04, 4.98±0.03 and 5.01±0.07, respectively. The serum total protein
concentration on day 5 and 6 was significantly (P<0.001) lower compared to control
group value of 5.96±0.06 and 5.85±0.10, respectively (Table 13 and Fig. 42).
The total protein concentration (g/dl) in Group VI birds administered with
ketoprofen (4 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 5.03±0.04, 5.36±0.08,
5.35±0.07, 5.53±0.06, 5.38±0.16 and 5.16±0.11, respectively. The serum total protein
concentration on day 5 and 6 was significantly (P<0.001) lower compared to control
group value of 5.96±0.06 and 5.85±0.10, respectively (Table 13 and Fig. 42).
The total protein concentration (g/dl) in Group VII birds administered with
meloxicam (0.5 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 4.93±0.12, 5.28±0.07,
5.31±0.04, 5.40±0.04, 5.76±0.08 and 5.23±0.10, respectively. The serum total protein
concentration on day 6 was significantly (P<0.01) lower compared to control group value
of 5.85±0.10, respectively (Table 13 and Fig. 42).
Other treated groups did not show any significant (P>0.05) alterations in the
values of serum total protein compared to the control group values.
4.7.9 Serum sodium concentration (meq/L)
The serum sodium concentration (meq/L) in Group II birds administered with
diclofenac (2. 5 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 155.40±1.30, 149.20±2.50,
160.70±2.40, 180.20±1.70, 200.10±2.90 and 220.30±3.00, respectively. Except for days
1, 2 and 3 serum sodium concentration for all other days (4, 5 and 6) was significantly
120
(P<0.001) higher compared to control group values of 155.30±1.40, 160.40±1.80 and
155.20±1.70, respectively (Table 14 and Fig. 43).
The serum sodium concentration (meq/L) in Group III birds administered with
aspirin (10 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 150.40±1.70, 156.90±1.40,
155.40±1.00, 153.10±2.10, 160.20±1.90 and 167.00±1.70, respectively. The serum
sodium concentration (meq/L) on day 6 was significantly (P<0.001) higher compared to
control group values (Table 14 and Fig. 43).
The serum sodium concentration (meq/L) in Group IV birds administered with
paracetamol (10 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 155.40±2.10, 154.20±3.40
159.30±2.90, 152.40±2.10, 160.20±2.00 and 164.20±1.90, respectively. The serum
sodium concentration (meq/L) on day 6 was significantly (P<0.05) higher compared to
control group values (Table 14 and Fig. 43).
The serum sodium concentration (meq/L) in Group V birds administered with
nimesulide (2 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 150.30±1.00, 152.60±1.60,
160.20±1.80, 150.20±2.00, 165.30±2.40, 167.00±2.60, respectively. The serum sodium
concentration (meq/L) on day 6 was significantly (P<0.001) higher compared to control
group values (Table 14 and Fig. 43).
The serum sodium concentration (meq/L) in Group VI birds administered with
ketoprofen (4 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 155.20±1.30, 152.30±1.50,
157.20±1.10, 160.30±2.30, 155.90±1.50 and 164.20±1.60, respectively. The serum
sodium concentration (meq/L) on day 6 was significantly (P<0.05) higher compared to
control group values (Table 14 and Fig. 43).
Table 13: Effect of oral administration of NSAIDs on serum total protein concentration (mg/dl) in broiler chickens
Total protein (g/dl)
Groups
Day
1
2
3
4
5
6
4.66±0.19
5.21±0.14
5.03±0.27
5.18±0.18
5.96±0.06
5.85±0.10
Group II
(Diclofenac)
4.95±0.09
4.56±0.24**
4.38±0.09**
n=5
4.20±0.06***
n=3
3.90±0.10***
n=2
3.85±0.00***
n=2
Group III
(Aspirin)
4.66±0.06
5.18±0.15
5.08±0.11
5.21±0.21
5.18±0.06***
5.01±0.16***
Group IV
(Paracetamol)
4.75±0.15
4.86±0.12
5.13±0.15
5.11±0.06
5.30±0.03**
5.08±0.15***
Group V
(Nimesulide)
4.95±0.14
5.00±0.15
5.33±0.07
5.16±0.04
4.98±0.03***
5.01±0.07***
Group VI
(Ketoprofen)
5.03±0.04
5.36±0.08
5.35±0.07
5.53±0.06
5.38±0.16**
5.16±0.11**
Group VII
(Meloxicam)
4.93±0.12
5.28±0.07
5.31±0.04
5.40±0.04
5.76±0.08
5.42±0.10
Group VIII
(Celecoxib)
5.06±0.18
5.58±0.11
5.20±0.08
5.28±0.19
5.85±0.14
5.71±0.09
Values are Mean+SE ; For each group n=6 unless otherwise mentioned
*P<0.05, **P<0.01, ***P<0.001 in relation to control.
121
Group I
(Control)
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
5
4
122
Total protein concentration (g/dl)
6
3
2
1
0
1
2
3
4
5
6
Day
Fig. 42: Effect of oral administration of NSAIDs on serum total protein concentration (g/dl) in broiler chickens
Table 14: Effect of oral administration of NSAIDs on serum sodium concentration (meq/L) in broiler chickens
Serum Na concentration (meq/L)
Groups
Day
1
Group I
(Control)
2
3
4
5
6
150.60±1.00
160.30±2.00
155.30±1.40
160.40±1.80
155.20±1.70
Group II
(Diclofenac)
155.40±1.30
149.20±2.50
160.70±2.40
n=5
180.20±1.70***
n=3
200.10±2.90***
n=2
220.30±3.00***
n=2
Group III
(Aspirin)
150.40±1.70
156.90±1.40
155.40±1.00
153.10±2.10
160.20±1.90
167.00±1.70***
Group IV
(Paracetamol)
155.40±2.10
154.20±3.40
159.30±2.90
152.40±2.10
160.20±2.00
164.20±1.90*
Group V
(Nimesulide)
150.30±1.00
152.60±1.60
160.20±1.80
150.20±2.00
165.30±2.40
167.00±2.60***
Group VI
(Ketoprofen)
155.20±1.30
152.30±1.50
157.20±1.10
160.30±2.30
155.90±1.50
164.20±1.60*
Group VII
(Meloxicam)
155.90±2.10
152.30±2.50
162.10±1.40
157.20±1.60
160.40±2.70
154.10±2.30
Group VIII
(Celecoxib)
149.20±2.30
150.30±2.50
154.20±2.40
154.30±3.00
160.30±3.10
157.20±3.00
Values are Mean+SE; For each group n=6 unless otherwise mentioned
*P<0.05, ***P<0.001 in relation to control.
123
149.20±1.20
225
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
175
150
125
124
Serum Na concentraion (meq/L)
200
100
75
50
25
0
1
2
3
4
5
6
Day
Fig. 43: Effect of oral administration of NSAIDs on serum Na concentration (meq/L) in broiler chickens
125
Other treated groups did not show any significant (P>0.05) alterations in the
values of serum sodium concentration compared to the control group values.
4.7.10 Serum potassium concentration (meq/L)
The serum potassium concentration (meq/L) in Group II birds administered with
diclofenac (2.5 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 4.50±0.10, 4.62±0.13,
6.21±0.14, 10.23±0.21, 14.21±0.29 and 16.13±0.30, respectively. Except for day 1 and 2
serum potassium concentration for all other days (3, 4, 5 and 6) was significantly
(P<0.001) higher compared to control group values (Table 14 and Fig. 44).
The serum potassium concentration (meq/L) in Group III birds administered with
aspirin (10 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 4.01±0.17, 4.21±0.14, 4.11±0.10,
4.65±0.13, 5.81±0.16 and 6.78±0.14, respectively. The serum potassium concentration
(meq/L) on day 5 and 6 was significantly (P<0.001) higher compared to control group
values (Table 14 and Fig. 44).
The serum potassium concentration (meq/L) in Group IV birds administered with
paracetamol (10 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 4.10±0.14, 4.12±0.13,
4.03±0.15, 4.54±0.13, 4.83±0.12 and 5.94±0.14, respectively. The serum potassium
concentration (meq/L) on day 6 was significantly (P<0.05) higher compared to control
group values (Table 14 and Fig. 44).
The serum potassium concentration (meq/L) in Group V birds administered with
nimesulide (2 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 4.52±0.12, 4.65±0.10,
4.21±0.13, 4.74±0.14, 5.22±0.13 and 6.72±0.15, respectively. The serum potassium
concentration (meq/L) on day 5 and 6 was significantly (P<0.01) higher compared to
control group values (Table 15 and Fig. 44).
126
The serum potassium concentration (meq/L) in Group VI birds administered with
ketoprofen (4 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 4.17±0.13, 4.27±0.12,
4.62±0.12, 4.40±0.14, 5.20±0.15 and 5.94±0.14, respectively. The serum potassium
concentration (meq/L) on day 5 and 6 was significantly (P<0.01) higher compared to
control group values (Table 15 and Fig. 44).
The serum potassium concentration (meq/L) in Group VII birds administered with
meloxicam (0.5 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 4.34±0.12, 4.54±0.10,
4.61±0.13, 4.54±0.14, 4.79±0.10 and 5.72±0.13, respectively. The serum potassium
concentration (meq/L) on day 6 was significantly (P<0.001) higher compared to control
group values (Table 15 and Fig. 44).
The serum potassium concentration (meq/L) in Group VIII birds administered
with celecoxib (3.5 mg/kg, PO) on days 1, 2, 3, 4, 5 and 6 was 4.55±0.15, 4.65±0.13,
4.40±0.14, 4.52±0.16, 4.87±0.17 and 5.79±0.10, respectively. The serum potassium
concentration (meq/L) on day 6 was significantly (P<0.001) higher compared to control
group values (Table 15 and Fig. 44).
Other treated groups did not show any significant (P>0.05) alterations in the
values of serum potassium concentration compared to the control group values.
Table 15: Effect of oral administration of NSAIDs on serum potassium concentration (meq/L) in broiler chickens
Serum K concentration (meq/L)
Groups
Day
1
2
3
4
5
6
4.51±0.12
4.43±0.10
4.52±0.14
4.64±0.10
4.64±0.15
4.71±0.14
Group II
(Diclofenac)
4.50±0.10
4.62±0.13
6.21±0.14***
n=5
10.23±0.21***
n=3
14.21±0.29***
n=2
16.13±0.30***
n=2
Group III
(Aspirin)
4.01±0.17
4.21±0.14
4.11±0.10
4.65±0.13
5.81±0.16***
6.78±0.14***
Group IV
(Paracetamol)
4.10±0.14
4.12±0.13
4.03±0.15
4.54±0.13
4.83±0.12
5.94±0.14***
Group V
(Nimesulide)
4.52±0.12
4.65±0.10
4.21±0.13
4.74±0.14
5.22±0.13**
6.72±0.15***
Group VI
(Ketoprofen)
4.17±0.13
4.27±0.12
4.62±0.12
4.40±0.14
5.20±0.15**
5.94±0.14***
Group VII
(Meloxicam)
4.34±0.12
4.54±0.10
4.61±0.13
4.54±0.14
4.79±0.10
5.72±0.13***
Group VIII
(Celecoxib)
4.55±0.15
4.65±0.13
4.40±0.14
4.52±0.16
4.87±0.17
5.79±0.10***
Values are Mean+SE ; For each group n=6 unless otherwise mentioned
**P<0.01, ***P<0.001 in relation to control.
127
Group I
(Control)
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
12.5
10.0
128
Serum K concentration (meq/L)
15.0
7.5
5.0
2.5
0.0
1
2
3
4
5
6
Day
Fig. 44: Effect of oral administration of NSAIDs on serum pottasium concentration (meq/L) in broiler chickens
129
4.8 Organ to body weight ratio
The effect of oral administration of NSAIDs on organ to body weight ratio in
broiler chickens were calculated and presented in Table 16 and Figure 55.
The organ to body weight ratio of liver, kidney, heart and spleen in Group II birds
administered with diclofenac (2.5 mg/kg, PO) was 2.40±0.07, 0.58±0.03, 0.49±0.02 and
0.11±0.00, respectively. There was significant (P<0.001) increase in the organ to body
weight ratio of liver and kidney compared to control group values.
The organ to body weight ratio of liver, kidney, heart and spleen in Group III
birds administered with aspirin (10 mg/kg, PO) was 2.61±0.09, 0.68±0.02, 0.58±0.02 and
0.13±0.01, respectively. There was significant (P<0.001) increase in the organ to body
weight ratio of liver compared to control group values.
The organ to body weight ratio of liver, kidney, heart and spleen in Group VI
birds administered with ketoprofen (4 mg/kg, PO) was 2.32±0.03, 0.62±0.03, 0.58±0.01
and 0.17±0.01, respectively. There was significant (P<0.01) increase in the organ to body
weight ratio of liver compared to control group values.
The organ to body weight ratio of liver, kidney, heart and spleen in Group VIII
birds administered with celecoxib (3.5 mg/kg, PO) was 2.36±0.14, 0.52±0.00, 0.56±0.00
and 0.11±0.01, respectively. There was significant (P<0.01) increase in the organ to body
weight ratio of liver compared to control group values.
Other treated groups did not show any significant (P>0.05) alterations in the
values of organ to body weight ratio of liver, kidney, heart and spleen compared to the
control group values.
Table 16: Effect of oral administration of NSAIDs on organ to body weight ratio in broiler chickens
Organ/ Body weight ratio
Groups
Liver
Group I
(Control)
2.14±0.06
Kidney
Heart
Spleen
0.62±0.03
0.52±0.02
0.10±0.01
2.40±0.07***
0.813±0.04*
0.62±0.02*
0.11±0.00
Group III
(Aspirin)
2.61±0.09***
0.68±0.02
0.58±0.02
0.13±0.01
Group IV
(Paracetamol)
2.16±0.05
0.61±0.01
0.57±0.03
0.12±0.00
Group V
(Nimesulide)
2.14±0.07
0.58±0.01
0.50±0.00
0.10±0.00
Group VI
(Ketoprofen)
2.32±0.03**
0.62±0.03
0.58±0.01
0.17±0.01
Group VII
(Meloxicam)
2.16±0.07
0.54±0.02
0.52±0.02
0.11±0.00
Group VIII
(Celecoxib)
2.36±0.14**
0.52±0.00
0.56±0.00
0.11±0.01
Values are Mean+SE ; For each group n=6
*P<0.05, **P<0.01, ***P<0.001 in relation to control.
130
Group II
(Diclofenac)
Control
Diclofenac
Aspirin
Paracetamol
Nimesulide
Ketoprofen
Meloxicam
Celecoxib
2.0
1.5
131
Organ/Body weight Ratio
2.5
1.0
0.5
0.0
Liver
Kidney
Heart
Fig. 45: Effect of oral administration of NSAIDs on organ to body weight ratio in broiler chickens
Spleen
132
4.9 Pathology
4.9.1 Gross pathology
Post mortem examination of the birds treated with diclofenac revealed congested
musculature along with deposition of chalky white urates on the visceral organs (Plate 4).
The heart and pericardium showed diffuse deposition of chalky white material (Plate 5).
The liver was congested and friable with varying degrees of urate deposition on the
surface of liver (Plate 6). Kidneys were congested and considerably enlarged, bulging out
of the renal fossa exhibiting prominent lobulation. Multifocal chalky white urate crystals
were noticed on surface of the kidneys (Plate 7). Peticheal hemorrhages were observed on
proventricular mucosa (Plate 8) and on ceca (Plate 9). These birds also showed deposition
of chalky white urate crystals on condyles of hock joint (Plate 10).
Post mortem examination of the birds administered with aspirin revealed enlarged
papillae accompanied with varying degrees of erosions on the mucosa of proventriculus
(Plate 11). The intestine showed grossly severe congestion and hemorrhages on the
serosal surface (Plate 12). All other organs were apparently normal.
The birds administered with paracetamol grossly revealed presence of erosions or
formation of crater on the mucosa of proventriculus (Plate 13). Intestine showed severe
congestion (Plate 14). All other organs were apparently normal.
Post mortem examination of the birds administered with nimesulide revealed
slight erosions on the mucosa of proventriculus and intestine. All other organs were
apparently normal.
Other treated groups did not show gross morphological changes in any of the
organs examined.
Plate 4: Group II- bird showing congested musculature with
deposition of chalky white urates on the visceral organs.
Plate 5: Group II- bird showing diffused urate
deposition on the surface of liver and pericardium.
Plate 6: Group II – pericardium of heart showing considerable
thickening with diffuse deposition of chalky white material.
133
Plate 3: Group II- birds (in the upper slot) showing anorexia,
dullness, lethargy and recumbence.
Plate 8: Group II- bird showing peticheal hemorrhages
on ceca.
Plate 9: Group II- bird showing enlarged kidneys with
prominent lobulation and deposition of urate crystals.
Plate 10: Group II- bird showing chalky white crystals
on condyles of hock joint.
134
Plate 7: Group II- bird showing peticheal hemorrhages in
the proventricular mucosa.
Plate 12: Group III- bird showing severe congestion and
hemorrhages on the serosal surface of intestine.
Plate 13: Group IV- bird showing erosions on the mucosa
of proventriculus.
Plate 14: Group IV- bird showing congestion of intestine.
135
Plate 11: Group III- bird showing enlarged papillae and
erosions on the mucosa of proventriculus.
136
4.9.2 Histopathology
On microscopic examination of diclofenac treated group; proventriculus section
showed degeneration and necrosis of villus epithelium, hemorrhages in the submucosa
and lamina propria, epithelial hyalinization occompanied with infiltration of
inflammatory cells (Plate 15). DeGalantha’s stained section of proventriculus tissue
showed deposition of uric acid on the serosal surface of the proventriculus (Plate 16).
Section of intestine showed shortening of villi, hemorrhage, degeneration and
necrosis of villus epithelium with infiltration of lymphoid cells in the lamina propria
(Plate 17 & 18).
Kidney section showed vascular and degenerative changes along with urate
deposition. Focal or multifocal areas of tubular epithelial degeneration and necrosis
accompanied with inflammatory changes (Plate 19). Tubular atrophy and presence of
proteinaceous material along with vacular degeneration was also observed in tubular
epithelial cells of some tubules. Black colour stained uric acid crystals were prominently
observed in kidney tubules in DeGalantha’s stain (Plate 20).
The liver section showed congestion of sinusoids and vessels. Hepatocyte
degeneration and necrosis accompanied with infiltrations of inflammatory cells (Plate
21). Deposition of black stained uric acid crystals in rosette pattern with occasional focal
irregular black spots were significantly observed in the DeGalantha’s stained sections of
liver (Plate 22).
Kidney sections under electron microscope showed dilatation of glomerular tufts
with RBCs, presence of electron dense granular area at proximal portion of glomerular
137
tufts (Plate 23). Tubular epithelial vaculation with intact nucleus, granular cytoplasm and
absence of endoplasmic reticulum (Plate 24).
Liver sections under electron microscope showed distortion of hepatocytes,
dilatation of sinusoidal areas and vaculation of cytoplasam (Plate 25). Degenerating
nucleus with hazy nuclear boundries, presence of fat globules and electron dense material
in cytoplasam was observed (Plate 26).
The section of heart showed congestion, hemorrhage, sub-epicardial edema and
urate deposition with infiltration of inflammatory cells in between cardiac muscle fibers
(Plate 27). In addition to these, infiltration of inflammatory cells in the pericardium,
congestion and oedema of myocardium was also noticed (Plate 29). Focal myocarditis
characterized by loss of cross striations, fragmentation of myocardial fibers with
infiltration of inflammatory cells were noticed occasionally. Deposition of black stained
uric acid crystals in rosette pattern with focal irregular black spots were significantly
observed in the DeGalantha’s stained heart sections (Plate 28 & 30).
On microscopic examination of aspirin treated group, proventriculus section
showed hemorrhages in the submucosa or lamina propria, hyalinization of villus
epithelium occompanied with infiltration of inflammatory cells (Plate 31).
Section of intestine showed erosion and desquamation of of villus epithelium into
the lumen with infiltration of inflammatory cells (Plate 32).
Microscopic examination of paracetamol treated group, proventriculus section
showed villus degeneration and infiltration of inflammatory cells (Plate 33).
Plate 17: Group II-section of intestine showing shortening
of villi (H & E 100X)
Plate 18: Group II-section of intestine showing hemorhage,
degeneration and necrosis of villus epithelium (H & E 100X)
138
Plate 15: Group II- proventriculus section showing
Plate 16: Group II- black coloured uric acid on the serosal
degeneration and necrosis of villus epithelium. (H & E 100X)
surface of the proventriculus (DeGalantha’s 100X)
Plate 20: Group II-kidney sections showing black stained uric
acid crystals in kidney tubules (DeGalantha’s 100X)
Plate 21: Group II- liver section showing hepatocyte
Plate 22: Group II-liver section showing deposition of black
degeneration and necrosis with inflammatory cells (H & E 200X)
stained uric acid crystals in rosette pattern (DeGalantha’s 200X)
139
Plate 19:Group II- kidney sections showing tubular
necrosis with inflammatory changes (H & E 200X)
140
Plate 23: Group II-Kidney sections under electron microscope showing dilatation of
glomerular tufts with RBCs, presence of electron dense granular area at proximal portion
of glomerular tufts (8950X)
Plate 24: Group II-Kidney section showing tubular epithelial vaculation with intact
nucleus, granular cytoplasm and absence of endoplasmic reticulum (4475X)
141
Plate 25: Group II-Liver sections showing distortion of hepatocytes, presence of fat
globule, dilatation of sinusoidal areas and vaculation of cytoplasam (4475X)
Plate 26: Group II-Liver section showing degenerating nucleus with hazy nuclear
boundries, presence of electron dense material in cytoplasam (4320X)
142
Section of the intestine showed hyperplastic changes in the crypts, erosion and
desquamation of villus epithelium into the lumen with infiltration of inflammatory cells
(Plate 34).
Section of the liver showed slightly swollen hepatocyte with granular cytoplasm
and mild congestion of vessels as well as sinusoids. Periductular hepatocytes
degeneration and necrosis with massive infiltration of inflammatory cells were also
noticed (Plate 35).
Microscopic examination of nimesulide treated group; proventriculus section
showed increased secretory activity, desquamation and degeneration of villus epithelium
and infiltration of inflammatory cells in the sub mucosa. (Plate 36).
Section of the intestine showed hyperplastic changes in crypts with increased
goblet cell activity and infiltration of inflammatory cells (Plate 37).
Section of the liver showed degeneration and necrosis of hepatocytes around the
bile duct, hyperplasia of bile duct epithelium accompanied with massive infiltration of
inflammatory cells (Plate 38).
However, no apparent histopathological lesions were observed in any of the tissue
section of control birds and in birds treated with NSAIDs such as ketoprofen, meloxicam
and celecoxib.
Plate 28:Group II-heart showing deposition of black stained
uric acid crystals (DeGalantha’s 100X)
Plate 29: Group II- heart showing infiltration of inflammatory
cells in the pericardium (H & E 100X)
Plate 30: Group II-heart showing deposition of black stained
uric acid crystals on pericardium (DeGalantha’s 100X)
143
Plate 27:Group II- heart showing sub-epicardial edema with
infiltration of inflammatory cells (H & E 100X)
Plate 33: Group IV- proventriculus section showing
hemorrhage and villus degeneration (H & E 100X)
Plate 34: Group IV- intestine showing hyperplastic changes in
crypts, erosion of villi with inflammatory cells (H & E 100X)
144
Plate 31: Group III- proventriculus section showing hemorrhages Plate 32: Group III- section of intestine showing erosion of
in the lamina propria with inflammatory cells (H & E 100X)
villus epithelium with inflammatory cells (H & E 100X)
Plate 36: Group V- proventriculus section showing increased
secretion with hemorrhage in the sub mucosa (H & E 200X)
Plate 37: Group V- section of intestine showing hyper plastic
Plate 38: Group V- section of liver showing degeneration and
changes in crypts with increased goblet cell activity(H & E 100X) necrosis of hepatocytes around the bile duct (H & E 100X)
145
Plate 35: Group IV-section of liver showing periductular
hepatocyte degeneration and necrosis (H & E 100X)