IV. RESULTS 4.1 Ultraviolet spectroscopy 4.1.1 Ultraviolet...

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.

Fig. 1. UV-visible spectrum of diclofenac sodium

200 225 250 275 300 325 3500.0

0.5

1.0

1.5

2.0

Acidic SolventAlkaline SolventNeutral Solvent

279

275

273

Wavelength in nm

Abs

orba

nce

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.

Fig. 2. UV-visible spectrum of aspirin

200 225 250 275 3000.0

0.5

1.0

1.5

Acidic SolventNeutral Solvent

276

278

225

230

Wavelength in nm

Abs

orba

nce

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.

Fig. 3. UV- visible spectrum of paracetamol

200 225 250 275 300 3250.0

0.5

1.0

1.5

2.0


245 256

250

Wavelength in nm

Abs

orba

nce

63

4.1.4 Ultraviolet visible spectrum of nimesulide The ultraviolet visible spectrum (200 - 550 nm) of nimesulide in acidic (0.1 N


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


nimesulide.

Fig. 4. UV- visible spectrum of nimesulide

200 250 300 350 400 450 500 5500.0

0.5

1.0

1.5

2.0


301 400

394

Wavelength in nm

Abs

orba

nce

64

4.1.5 Ultraviolet visible spectrum of ketoprofen The ultraviolet visible spectrum (200 - 350 nm) of ketoprofen in acidic (0.1 N


The maximum ultraviolet absorption of ketoprofen was found at 260, 262 and 257 nm in


respectively. The obtained spectra and maximum absorption wavelength for ketoprofen


ketoprofen.

Fig. 5. UV- visible spectrum of ketoprofen

200 250 300 3500.0

0.5

1.0

1.5

2.0


260

262

Wavelength in nm

Abs

orba

nce

257

65

4.1.6 Ultraviolet visible spectrum of meloxicam The ultraviolet visible spectrum (200 - 400 nm) of meloxicam in acidic (0.1 N


The maximum ultraviolet absorption of meloxicam was found at 345, 362 and 356 nm in


respectively. The obtained spectra and maximum absorption wavelength for meloxicam


meloxicam.

Fig. 6. UV- visible spectrum of meloxicam

200 250 300 350 4000.0

0.5

1.0

1.5

2.0


345

362

356

Wavelength in nm

Abs

orba

nce

66

4.1.6 Ultraviolet visible spectrum of celecoxib The ultraviolet visible spectrum (200 - 350 nm) of celecoxib in acidic (0.1 N


The maximum ultraviolet absorption of celecoxib was found at 250, 252 and 254 nm in


respectively. The obtained spectra and maximum absorption wavelength for celecoxib in

acidic and alkaline solvent system was compared with the reference spectra of celecoxib.

Fig. 7. UV- visible spectrum of celecoxib

200 225 250 275 300 325 3500.0

0.5

1.0

1.5

2.0


250

252

254

Wavelength in nm

Abs

orba

nce

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

1194.25, 1166.11

1234.26, 1250.09

1282.16

C-H in plane bending aromatic

769.40, 747.72 714.78, 744.82 C-H out of plane bending-o-

disubstituted

1400-1500 &

1585-1600

1451.29, 1604.01 C=C stretching aromatic

68

715.

7174

7.72

769.

40

839.

6886

9.26

952.

0210

45.0

110

89.1

4

1199

.65

1249

.39

1283

.85

1305

.78

1393

.30

1453

.18

1470

.43

1508

.27

1555

.54

1574

.72

3387

.39

-0

10

20

30

40

50

60

70

80

90

100

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 8. Infrared spectrum of diclofenac sodium (KBr disk)

452.

6248

4.25

530.

31

558.

4663

3.60

714.

7874

4.82

766.

3784

0.00

868.

8795

1.92

1044

.77

1089

.19

1166

.11

1194

.25

1234

.26

1250

.09

1282

.16

1305

.08

1384

.39

1451

.29

1469

.21

1497

.41

1553

.24

1572

.80

1604

.01

3225

.86

-0

10

20

30

40

50

60

70

80

90

100

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 9. Infrared spectrum of diclofenac sodium (ATR)

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


1753.61

1748.91

C=O stretching- carboxylic acids

1220.82, 1306.84 1011.70, 1092.97,

1218.24, 1286.83

C-O stretching - esters

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,

1094.86, 1134.99,

1187.80, 1220.82

1011.70, 1092.97,

1218.24, 1286.83


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

70

666.

7270

4.84

755.

5279

0.71

804.

0984

0.28

917.

3297

0.41

1013

.08

1038

.88

1094

.86

1134

.99

1187

.80

1220

.8213

06.8

413

70.6

214

19.5

914

58.0

214

83.6

015

75.9

516

05.7

716

91.9

7

1753

.61

2699

.86

2921

.91

10

15

20

25

30

35

40

45

50

55

60

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 10. Infrared spectrum of aspirin (KBr disk)

423.

4051

4.31

541.

83

597.

2264

3.16

665.

78

802.

4283

8.38

914.

33

969.

84

1011

.70

1092

.97

1218

.24

1286

.83

1367

.98

1417

.81

1455

.74

1481

.99

1604

.38

1680

.72

1748

.91

2829

.67

50

55

60

65

70

75

80

85

90

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 11. Infrared spectrum of aspirin (ATR)

71

4.2.3 Infrared spectrum of parcetamol

The infrared spectrum of paracetamol in KBr disk and ATR is shown in Fig. 12


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


3325.87

3318.96

OH stretching phenolics

1655.79 1650.57 N-H bending

1226.16,1259.81,

1370.53

1224.43,

1258.16, 1370.52

C-O stretching phenolics

1015.16, 107.71,

1172.00, 226.61,

1243.18, 259.81

1015, 1107.37,

1171.56,

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

72

685.

1171

3.94

796.

2880

8.30

837.

1585

7.12

968.

4710

15.1

611

07.7

111

72.0

012

26.6

112

43.1

8

1259

.81

1327

.37

1370

.53

1442

.17

1506

.40

1564

.74

1610

.59

1655

.79

1877

.20

2586

.72

2793

.21

3161

.85

3325

.87

40

42

44

46

48

50

52

54

56

58

60

62

64

66

68

70

72

74

76

78

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 12. Infrared spectrum of paracetamol (KBr disk)

414.

3246

4.21

501.

26

517.

48

602.

8362

4.42

680.

88

711.

9279

5.82

806.

4983

5.78

857.

31

967.

91

1015

.08

1107

.37

1171

.56

1224

.43

1258

.16

1327

.64

1370

.52

1434

.05

1505

.10

1561

.931609

.71

1650

.57

3109

.1933

18.9

6

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 13. Infrared spectrum of paracetamol (ATR)

73

4.2.4 Infrared spectrum of nimesulide

The infrared spectrum of nimesulide in KBr disk and ATR is shown in Fig. 14



characteristic for nimesulide.

Wave number

(cm-1) KBr disk

Wave number

(cm-1) ATR


1343.79, 1152.0

-

S=O stretching - Sulfonamides

3281.83 3277.32 N-H stretching - Sulfonamides (10)

740.75, 753.23,

803.80, 830.71,

848.69

802.46, 830.15 N-O stretching - Nitrites

871.85 871.08 C-N stretching - Nitroaromatics

1018.99,1126.84,

1068.91,1080.06,

1217.48,1152.07,

1185.92,1247.91,

1282.18

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,

1588.51

1404.34, 1445.31,

1486.66, 1587.56

C=C stretching - Aromatic

74

663.

7669

8.07

740.

7575

3.23

803.

8083

0.71

848.

6987

1.85

905.

4295

0.64

976.

1710

18.9

9

1068

.91

1080

.06

1126

.84

1152

.07

1185

.97

1217

.48

1247

.91

1282

.18

1316

.31

1343

.79

1406

.41

1445

.31

1487

.87

1502

.90

1522

.45

1560

.30

1588

.51

1599

.48

1653

.90

1700

.97

1735

.33

2928

.91

3085

.39

3281

.83

3438

.86

3727

.44

54

56

58

60

62

64

66

68

70

72

74

76

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 14. Infrared spectrum of nimesulide (KBr disk)

445.

1047

8.56

512.

9755

1.19

696.

85740.

03802.

4683

0.15

871.

08

903.

71

949.

73

972.

98

1078

.95

1149

.50

1215

.49

1245

.50

1314

.51

1404

.34

1445

.31

1486

.66

1587

.56

3277

.32

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100

%T

500 1000 1500 2000 2500 3000 3500 Wavenumbers (cm-1)

Fig. 15. Infrared spectrum of nimesulide (ATR)

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


characteristic for ketoprofen.

Wave number

(cm-1) KBr disk

Wave number

(cm-1) ATR


1697.38

1654.50

C=O stretching ketones

1134.57, 1174.86,

1194.79, 1227.84,

1285.34

1134.14, 1174.57,

1195.28, 1227.32,

1282.64

C=O stretching and bending

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,

1174.86, 1194.79,

1227.84, 1285.34.

1077.65, 1134.14,

1174.57, 1195.28,

1227.32, 1282.64

C-H in plane bending - Aromatic

691.02, 703.53,

716.75, 773.10,

787.29

689.94, 703.13,

714.77, 787.17,

773.13

C-H out of plane bending-m-

disubstituted

1445.31, 1420.60,

1598.20

1420.61, 1441.26,

1597.68

C=C stretching aromatic

76

691.

0270

3.53

716.

75

773.

1078

7.29

811.

1782

6.75

866.

0791

5.97

968.

08

1078

.31

1134

.57

1174

.86

1194

.79

1227

.84

1285

.34

1319

.28

1369

.88

1420

.60

1445

.31

1575

.56

1598

.20

1655

.21

1697

.38

2978

.67

15

20

25

30

35

40

45

50

55

60

65

70

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 16. Infrared spectrum of ketoprofen (KBr disk)

440.

0651

3.66

613.

6864

2.07

689.

94

703.

13

714.

7777

3.13

787.

1781

1.27

826.

6991

5.92

966.

26

1077

.65

1134

.14

1174

.57

1195

.28

1227

.32

1282

.64

1318

.92

1369

.52

1420

.61

1441

.26

1597

.68

1654

.50

2978

.64

30

35

40

45

50

55

60

65

70

75

80

85

90

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 17. Infrared spectrum of ketoprofen (ATR)

77

4.2.6 Infrared spectrum of meloxicam

The infrared spectrum of meloxicam in KBr disk and ATR is shown in Fig. 18



characteristic for meloxicam.

Wave number

(cm-1) KBr disk

Wave number

(cm-1) ATR


1620

1549.03, 1616.69,

644.39, 711.40,

760.55

N-H bending - Amides (10)

3291.17 3281.43 O-H stretching - Phenols

1301.41, 1346.38,

1457.05, 1530.28,

1550.52

1343.41, 1446.92,

1549.03

Ring stretching - Heteroaromatics

1185.26, 1346.38

1343.41 C-O stretching for phenols

1043.78, 1064.56,

118.65, 1130.91,

1153.05, 1161.82,

1185.26

1042.82, 1151.12,

1262.47


713.54, 761.70,

784.78, 798.30

711.40, 760.55 C-H out of plane bending-m-

disubstituted

1457.05 1446.92 C=C stretching aromatic

78

677.

6671

3.54

729.

9576

1.70

784.

7879

8.30

826.

8684

5.03

855.

8194

0.82

1043

.78

1064

.56

1118

.65

1130

.91

1153

.05

1161

.82

1185

.26

1217

.57

1265

.21

1301

.41

1346

.38

1457

.05

1530

.28

1550

.52

1620

.66

3291

.17

20

25

30

35

40

45

50

55

60

65

70

75

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 18. Infrared spectrum of meloxicam (KBr disk)

454.

7352

6.69

564.

7560

7.41

644.

3971

1.40

760.

55

823.

0494

0.03

1042

.82

1151

.12

1262

.47

1343

.41

1446

.92

1549

.03

1616

.6932

81.4

3

0

10

20

30

40

50

60

70

80

90

100

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 19. Infrared spectrum of meloxicam (ATR)

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


characteristic for celecoxib.

Wave number

(cm-1) KBr disk

Wave number

(cm-1) ATR


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


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-

substituted

1446.64, 1473.65, 1498.58

1402.51, 1473.94, 1445.84, 1497.15, 1593.78

C=C stretching aromatic

80

761.

8879

2.33

801.

7884

6.22

907.

2498

1.63

1016

.33

1093

.86

1135

.35

1164

.84

1229

.98

1274

.78

1347

.72

1374

.31

1446

.64

1473

.65

1498

.58

1605

.98

1695

.42

3235

.17

3341

.93

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 20. Infrared spectrum of celecoxib (KBr disk)

421.

6744

1.66

499.

5451

1.32

530.

35

559.

9263

0.77

647.

6072

1.03

742.

11

760.

6279

1.14

801.

09844.

77

904.

8996

9.40

980.

86

1015

.93

1101

.68

1132

.63

1157

.53

1228

.50

1274

.23

1346

.02

1374

.13

1402

.51

1445

.84

1473

.94

1497

.15

1561

.01

1593

.78

3096

.22

3226

.12

3332

.02

-5

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

%T

500 1000 1500 2000 2500 3000 3500 4000 Wavenumbers (cm-1)

Fig. 21. Infrared spectrum of celecoxib (ATR)

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

84

Fig 24:HPTLC-UV absorption spectra of diclofenac Fig 25: HPTLC-UV absorption spectra of paracetamol

Fig 26: HPTLC-UV absorption spectra of ketoprofen Fig 27: HPTLC-UV absorption spectra of celecoxib

200 250 300 350 4000

25

50

75

100 281

Wavelength in nm

Abs

orpt

ion

Uni

t

200 250 300 350 4000

25

50

75

100 246

Wavelength in nm

Abs

orpt

ion

Uni

t

200 250 300 350 4000

25

50

75

100 263

Wavelength in nm

Abs

orpt

ion

Uni

t

200 250 300 350 4000

25

50

75

100 258

Wavelength in nm

Abs

orpt

ion

Uni

t

85

Fig 28: HPTLC-UV absorption spectra of nimesulide Fig 29: HPTLC-UV absorption spectra of meloxicam Fig 30: HPTLC-UV absorption spectra of aspirin

200 250 300 350 400 450 5000

25

50

75

100 307

Wavelength in nm

Abs

orpt

ion

Uni

t

200 250 300 350 400 4500

25

50

75

100 361

Wavelength in nm

Abs

orpt

ion

Uni

t

200 250 300 350 4000

25

50

75

100

225

Wavelength in nm

Abs

orpt

ion

Uni

t

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 88

Table 2: Effect of oral administration of NSAIDs on RBC count (106/l) in broiler chickens

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.

RBC count (106/l) Groups

Day

1 2 3 4 5 6

Group I (Control) 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

89 89

Fig. 31: Effect of oral administration of NSAIDs on RBC count (106/l) in broiler chickens

1 2 3 4 5 60.0

0.5

1.0

1.5

2.0

2.5

3.0

ControlDiclofenacAspirinParacetamolNimesulideKetoprofenMeloxicamCelecoxib

Day

RB

C c

ount

(106 /l)

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


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.

91

Table 3: Effect of oral administration of NSAIDs on hemoglobin concentration (g/dl) in broiler chickens

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.

Hemoglobin concentration (g/dl) Groups

Day

1 2 3 4 5 6

Group I (Control) 13.01±0.12 13.00±0.09 12.89±0.08 12.60±0.13 12.56±0.12 12.50±0.11


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***


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**




92

Fig. 32: Effect of oral administration of NSAIDs on hemoglobin concentration (g/dl) in broiler chickens

1 2 3 4 5 60

1

2

3

4

5

6

7

8

9

10

11

12

13


Day

Hem

oglo

bin

conc

entr

atio

n (g

/dl)

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


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).


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


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


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


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


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,


The platelet count (×103/l) in Group VII birds administered with meloxicam (0.5


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,


The platelet count (×103/l) in Group VIII birds administered with celecoxib (3.5


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).


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


96

Table 4: Effect of oral administration of NSAIDs on PCV (%) in broiler chickens

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.

PCV (%) Groups

Day

1 2 3 4 5 6

Group I (Control) 33.12±0.21 33.00±0.23 32.80±0.30 32.71±0.14 32.00±0.31 31.53±0.21


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**


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**




97

Fig. 33 : Effect of oral administration of NSAIDs on PCV (%) in broiler chickens

1 2 3 4 5 60

5

10

15

20

25

30


Day

PCV

(%)

98

Table 5: Effect of oral administration of NSAIDs on platelet count (103/l) in broiler chickens

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.

Platelet count (103/l) Groups

Day

1 2 3 4 5 6

Group I (Control) 21±1.23 22±1.30 20±2.01 20±1.65 21±1.42 19±1.84


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***


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*

99

Fig. 34: Effect of oral administration of NSAIDs on platelet count (103/l) in broiler chickens

1 2 3 4 5 60

5

10

15

20

25


Day

Plat

elet

cou

nt (1

03 / l)

100

The AST concentration (U/L) in Group V birds administered with nimesulide (2


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



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


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


The ALT concentration (U/L) in Group V birds administered with nimesulide (2


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



values of ALT compared to the control group values.

101

Table 6: Effect of oral administration of NSAIDs on aspartate aminotransferase (AST) concentration in broiler chickens

Values are Mean±SE; For each group n=6 unless otherwise mentioned **P<0.01, ***P<0.001 in relation to control.

Asparatae aminotransferase (U/L) Groups

Day

1 2 3 4 5 6

Group I (Control) 78.35±4.83 82.69±5.87 97.80±0.71 98.67±0.54 97.72±2.58 99.84±2.96


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


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**




102

Fig. 35: Effect of oral administration of NSAIDs on serum AST concentration (U/L) in broiler chickens

1 2 3 4 5 60

25

50

75

100

125

150

175

200

225

250


Day

AST

con

cent

ratio

n (U

/L)

103

Table 7: Effect of oral administration of NSAIDs on alanine aminotransferase (ALT) concentration in broiler chickens

Values are Mean±SE; For each group n=6 unless otherwise mentioned **P<0.01, ***P<0.001 in relation to control.

Alanine aminotransferase (U/L) Groups

Day

1 2 3 4 5 6

Group I (Control) 19.63±0.29 19.25±1.39 19.87±1.04 20.78±0.53 21.17±0.52 20.72±0.41


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


18.76±2.89 18.58±1.32 20.34±1.15 21.90±0.84 21.99±0.97 25.48±3.06





104

Fig. 36 : Effect of oral administration of NSAIDs on serum ALT concentration (U/L) in broiler chickens

1 2 3 4 5 60

25

50

75


Day

ALT

con

cent

ratio

n (U

/L)

105

4.7.3 Alkaline phosphatase (ALP) concentration

The ALP concentration (KA units/dl) in Group II birds administered with


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,


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



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,




106

Table 8: Effect of oral administration of NSAIDs on alkaline phosphatase (ALP) concentration in broiler chickens

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.

Alkaline phosphatase (KA units/dl) Groups

Day

1 2 3 4 5 6

Group I (Control) 15.00±0.51 15.43±0.42 15.33±0.42 16.16±0.79 17.66±0.33 17.33±0.61


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


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*



107

Fig. 37: Effect of oral administration of NSAIDs on serum ALP concentration (KA units/dl) in broiler chickens

1 2 3 4 5 60

10

20

30


Day

ALP

con

cent

ratio

n (K

A u

nits

/dl)

108


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



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


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,


The creatinine concentration (mg/dl) in Group V birds administered with


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


109

Table 9: Effect of oral administration of NSAIDs on BUN concentration (mg/dl) in broiler chickens

Values are Mean±SE; For each group n=6 unless otherwise mentioned ***P<0.001 in relation to control.

Blood Urea Nitrogen (mg/dl) Groups

Day

1 2 3 4 5 6

Group I (Control) 3.66±0.16 3.51±0.12 3.78±0.13 3.85±0.20 3.95±0.20 4.75±0.08


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


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




110

Fig. 38: Effect of oral administration of NSAIDs on BUN concentration (mg/dl) in broiler chicken

1 2 3 4 5 60

1

2

3

4

5

6

7

8 ControlDiclofenacAspirinPracetamolNimesulideKetoprofenMeloxicamCelecoxib

Day

BU

N c

once

ntra

tion

(mg/

dl)

111

Table 10: Effect of oral administration of NSAIDs on serum creatinine concentration (mg/dl) in broiler chickens

Values are Mean+SE ; For each group n=6 unless otherwise mentioned **P<0.01, ***P<0.001 in relation to control.

Creatinine (mg/dl) Groups

Day

1 2 3 4 5 6

Group I (Control) 0.38±0.02 0.43±0.03 0.37±0.03 0.42±0.02 0.42±0.03 0.42±0.03


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


0.40±0.03 0.35±0.01 0.38±0.01 0.44±0.03 0.41±0.03 0.46±0.03





112

Fig. 39: Effect of oral administration of NSAIDs on serum creatinine concentration (mg/dl) in broiler chickens

1 2 3 4 5 60.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9ControlDiclofenacAspirinParacetamolNimesulideKetoprofenMeloxicamCelecoxib

Day

Cre

atin

ine

conc

entr

atio

n (m

g/dl

)

113

4.7.6 Uric acid concentration (mg/dl)

The uric acid concentration (mg/dl) in Group II birds administered with


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


The uric acid concentration (mg/dl) in Group VI birds administered with


4.23±0.06, 4.16±0.10, 4.51±0.04 and 5.31±0.14, respectively. The serum uric acid


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


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


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



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


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


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


115

Table 11: Effect of oral administration of NSAIDs on serum uric acid concentration (mg/dl) in broiler chickens

Values are Mean+SE ; For each group n=6 unless otherwise mentioned *P<0.05, ***P<0.001 in relation to control.

Uric acid (mg/dl) Groups

Day

1 2 3 4 5 6

Group I (Control) 4.10±0.15 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


3.83±0.03 4.00±0.05 4.05±0.07 4.40±0.05 4.60±0.01 5.00±0.09





116

Fig. 40: Effect of oral administration of NSAIDs on serum uric acid concentration (mg/dl) in broiler chickens

1 2 3 4 5 60

1

2

3

4

5

6

7

8

9

10 ControlDiclofenacAspirinParacetamolNimesulideKetoprofenMeloxicamCelecoxib

Day

Uri

c ac

id c

once

ntra

tion

(mg/

dl)

117

Table 12: Effect of oral administration of NSAIDs on serum albumin concentration (g/dl) in broiler chickens

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.

Albumin (g/dl) Groups

Day

1 2 3 4 5 6

Group I (Control) 1.85±0.06 1.96±0.16 1.95±0.07 1.96±0.08 2.10±0.10 2.13±0.12


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**


1.65±0.07 1.90±0.11 1.95±0.05 2.06±0.13 2.03±0.06 2.35±0.11





118

Fig. 41: Effect of oral administration of NSAIDs on serum albumin concentration (g/dl) in broiler chickens

1 2 3 4 5 60.0

0.5

1.0

1.5

2.0

2.5


Day

Alb

umin

con

cent

ratio

n (g

/dl)

119

The total protein concentration (g/dl) in Group V birds administered with



concentration on day 5 and 6 was significantly (P<0.001) lower compared to control


The total protein concentration (g/dl) in Group VI birds administered with



concentration on day 5 and 6 was significantly (P<0.001) lower compared to control


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,


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).


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


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




The serum sodium concentration (meq/L) in Group V birds administered with


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





121

Table 13: Effect of oral administration of NSAIDs on serum total protein concentration (mg/dl) in broiler chickens

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.

Total protein (g/dl) Groups

Day

1 2 3 4 5 6

Group I (Control)

4.66±0.19 5.21±0.14 5.03±0.27 5.18±0.18 5.96±0.06 5.85±0.10


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***


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

122

Fig. 42: Effect of oral administration of NSAIDs on serum total protein concentration (g/dl) in broiler chickens

1 2 3 4 5 60

1

2

3

4

5

6


Day

Tota

l pro

tein

con

cent

ratio

n (g

/dl)

123

Table 14: Effect of oral administration of NSAIDs on serum sodium concentration (meq/L) in broiler chickens

Values are Mean+SE; For each group n=6 unless otherwise mentioned *P<0.05, ***P<0.001 in relation to control.

Serum Na concentration (meq/L) Groups

Day

1 2 3 4 5 6

Group I (Control) 149.20±1.20 150.60±1.00 160.30±2.00 155.30±1.40 160.40±1.80 155.20±1.70


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***


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***




124

Fig. 43: Effect of oral administration of NSAIDs on serum Na concentration (meq/L) in broiler chickens

1 2 3 4 5 60

25

50

75

100

125

150

175

200

225ControlDiclofenacAspirinParacetamolNimesulideKetoprofenMeloxicamCelecoxib

Day

Seru

m N

a co

ncen

trai

on (m

eq/L

)

125


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


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



The serum potassium concentration (meq/L) in Group V birds administered with



concentration (meq/L) on day 5 and 6 was significantly (P<0.01) higher compared to


126

The serum potassium concentration (meq/L) in Group VI birds administered with



concentration (meq/L) on day 5 and 6 was significantly (P<0.01) higher compared to


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,




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,





values of serum potassium concentration compared to the control group values.

127

Table 15: Effect of oral administration of NSAIDs on serum potassium concentration (meq/L) in broiler chickens

Values are Mean+SE ; For each group n=6 unless otherwise mentioned **P<0.01, ***P<0.001 in relation to control.

Serum K concentration (meq/L) Groups

Day

1 2 3 4 5 6

Group I (Control)

4.51±0.12 4.43±0.10 4.52±0.14 4.64±0.10 4.64±0.15 4.71±0.14


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***


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***

128

Fig. 44: Effect of oral administration of NSAIDs on serum pottasium concentration (meq/L) in broiler chickens

1 2 3 4 5 60.0

2.5

5.0

7.5

10.0

12.5

15.0


Day

Seru

m K

con

cent

ratio

n (m

eq/L

)

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


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



values of organ to body weight ratio of liver, kidney, heart and spleen compared to the

control group values.

130

Table 16: Effect of oral administration of NSAIDs on organ to body weight ratio in broiler chickens

Values are Mean+SE ; For each group n=6 *P<0.05, **P<0.01, ***P<0.001 in relation to control.

Organ/ Body weight ratio Groups

Liver Kidney Heart Spleen

Group I (Control) 2.14±0.06 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


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

131

Fig. 45: Effect of oral administration of NSAIDs on organ to body weight ratio in broiler chickens

Liver Kidney Heart Spleen0.0

0.5

1.0

1.5

2.0

2.5


Org

an/B

ody

wei

ght R

atio

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.

133

Plate 3: Group II- birds (in the upper slot) showing anorexia, Plate 4: Group II- bird showing congested musculature with dullness, lethargy and recumbence. deposition of chalky white urates on the visceral organs. Plate 5: Group II- bird showing diffused urate Plate 6: Group II – pericardium of heart showing considerable

deposition on the surface of liver and pericardium. thickening with diffuse deposition of chalky white material.

134

Plate 7: Group II- bird showing peticheal hemorrhages in Plate 8: Group II- bird showing peticheal hemorrhages the proventricular mucosa. on ceca. Plate 9: Group II- bird showing enlarged kidneys with Plate 10: Group II- bird showing chalky white crystals prominent lobulation and deposition of urate crystals. on condyles of hock joint.

135

Plate 11: Group III- bird showing enlarged papillae and Plate 12: Group III- bird showing severe congestion and

erosions on the mucosa of proventriculus. hemorrhages on the serosal surface of intestine. Plate 13: Group IV- bird showing erosions on the mucosa Plate 14: Group IV- bird showing congestion of intestine.

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).

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 17: Group II-section of intestine showing shortening Plate 18: Group II-section of intestine showing hemorhage, of villi (H & E 100X) degeneration and necrosis of villus epithelium (H & E 100X)

139

Plate 19:Group II- kidney sections showing tubular Plate 20: Group II-kidney sections showing black stained uric

necrosis with inflammatory changes (H & E 200X) 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)

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)

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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)

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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.

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Plate 27:Group II- heart showing sub-epicardial edema with Plate 28:Group II-heart showing deposition of black stained infiltration of inflammatory cells (H & E 100X) uric acid crystals (DeGalantha’s 100X) Plate 29: Group II- heart showing infiltration of inflammatory Plate 30: Group II-heart showing deposition of black stained

cells in the pericardium (H & E 100X) uric acid crystals on pericardium (DeGalantha’s 100X)

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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 33: Group IV- proventriculus section showing Plate 34: Group IV- intestine showing hyperplastic changes in

hemorrhage and villus degeneration (H & E 100X) crypts, erosion of villi with inflammatory cells (H & E 100X)

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Plate 35: Group IV-section of liver showing periductular Plate 36: Group V- proventriculus section showing increased

hepatocyte degeneration and necrosis (H & E 100X) 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)

IV. RESULTS 4.1 Ultraviolet spectroscopy 4.1.1 Ultraviolet...

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