CLINICAL STUDIES ON ANAL AND PERINEAL TUMORS IN DOGS BY S.KOKILA B.V.Sc. & A.H. Thesis submitted to the SRI VENKATESWARA VETERINARY UNIVERSITY In partial fulfilment of the requirements For the award of the degree of MASTER OF VETERINARY SCIENCE IN THE FACULTY OF VETERINARY SCIENCE (VETERINARY SURGERY AND RADIOLOGY) DEPARTMENT OF VETERINARY SURGERY AND RADIOLOGY COLLEGE OF VETERINARY SCIENCE SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI – 517 502 SEPTEMBER 2011 i CERTIFICATE Dr. S.KOKILA has satisfactorily prosecuted the course of research and that the thesis entitled “CLINICAL STUDIES ON ANAL AND PERINEAL TUMORS IN DOGS ” submitted is the result of original research work and is of sufficiently high standard to warrant its presentation to the examination. I also certify that the thesis or part there of has not been previously submitted by her for a degree of any university. Date : (Dr. P.VEENA) Major Advisor Place : Tirupati Assistant Professor (s.s) Dept. of Veterinary Surgery & Radiology College of Veterinary Science Tirupati – 517 502 (A.P) ii CERTIFICATE This is to certify that the thesis entitled “CLINICAL STUDIES ON ANAL AND PERINEAL TUMORS IN DOGS” submitted in partial fulfilment of the requirements for the degree of Master of Veterinary Science of Sri Venkateswara Veterinary University, Tirupati is a record of the bonafide research work carried out by Dr.S.KOKILA under our guidance and supervision. The subject of the thesis has been approved by the Student’s Advisory Committee. No part of the thesis has been submitted for any other degree or diploma. The published part has been fully acknowledged. All the assistance and help received during the course of investigation have been duly acknowledged by the author of the thesis. (Dr. P.VEENA) Chairman of the Advisory Committee Thesis approved by the Student Advisory Committee CHAIRMAN: (Dr. P.VEENA) ________________________ Assistant Professor (s.s) Dept. of Veterinary Surgery & Radiology College of Veterinary Science Tirupati – 517 502 (A.P) MEMBER : (Dr. R.V. SURESH KUMAR) ________________________ Associate Professor and Head Dept. of Veterinary Surgery & Radiology College of Veterinary Science Tirupati – 517 502 (A.P) MEMBER : (Dr. Ch. SRILATHA ) _______________________ Professor & University Head Department of Veterinary Pathology, College of Veterinary Science Tirupati – 517 502 (A.P) iii LIST OF CONTENTS Chapter TITLE Page No. No. I INTRODUCTION 1 II REVIEW OF LITERATURE 6 2.1 Incidence of tumors 7 2.1.1 Age 7 2.1.2 Breed 8 2.1.3 Sex 9 2.2 Clinical symptoms 11 2.3 Diagnosis of tumors 12 2.3.1 Gross examination 12 2.3.1.1 Anal tumors 12 2.3.1.2 Perineal tumors 14 2.3.2 Cytological examination 17 2.3.3 Radiography 19 2.4 Treatment 20 2.4.1 Surgical excision 20 2.4.2 Chemotherapy 21 2.4.3 Other treatments 23 2.5 Complications 24 2.6 Parameters studied 25 2.6.1 Haematological observations 25 2.6.2 Biochemical observations 26 2.7 Pathological examination 27 2.7.1 Histopathology 27 2.7.1.1 Anal tumors 27 2.7.1.2 Perineal tumors 29 2.7.2 AgNOR staining and counting 31 2.7.3 Immunohistochemistry 32 iv 2.8 Recurrence, disease free intervals and prognosis 32 III MATERIALS AND METHODS 3.1 Materials 34 3.2 Methods 34 3.2.1 Incidence and clinical examination 34 3.2.2 Selection of cases 34 3.3 Design of the study 35 3.4 Diagnosis 36 3.4.1 Gross examination 36 3.4.2 Cytological examination 36 3.4.2.1 Fine needle aspiration biopsy (FNAB) 36 3.4.3 Radiographic studies 36 3.5 Surgical treatment 39 3.5.1 Patient preparation 39 3.5.2 Anaesthesia and control 39 3.5.3 Operative technique 39 3.5.4 Post operative care 40 3.6 Chemotherapy 40 3.6.1 Supportive therapy 40 3.7 Follow up studies 42 3.7.1 Post operative complications 42 3.7.2 Chemotherapy complications 42 3.8 Parameters studied 42 3.8.1 Physiological parameters 42 3.8.2 Hematology 42 3.8.2.1 Total leukocyte count (TLC) 42 3.8.2.2 Haemoglobin (Hb) 43 3.8.2.3 Packed cell volume (PCV) 43 3.8.2.4 Differential leukocyte count (DLC) 43 v 3.8.3 Serum biochemistry 43 3.8.3.1 Calcium 43 3.8.3.2 Phosphorus 43 3.8.3.3 Blood urea nitrogen 45 3.8.3.4 Creatinine 45 3.8.4 Pathological observations 45 3.8.4.1 Gross pathological observations 45 3.8.4.2 Histopathology 45 3.8.4.3 AgNOR staining 45 3.8.4.4 AgNOR counting 46 3.8.5 Immunohistochemistry 46 3.8.5.1 Reagents used 46 3.8.5.2 Procedure 47 3.8.5.3 Evaluation of hormone receptor expression 48 3.8.6 Recurrence, disease free intervals and prognosis 49 3.8.7 Statistical analysis 49 IV RESULTS 50 4.1 Incidence 50 4.1.1 Age 50 4.1.2 Breed 52 4.1.3 Sex 52 4.2 Clinical symptoms 56 4.3 Diagnosis 58 4.3.1 Gross examination 58 4.3.2 Cytological examination 58 4.3.2.1 Fine needle aspiration biopsy (FNAB) 58 4.3.3 Radiological studies 60 4.4 Treatment 60 4.4.1 Follow up studies 65 vi 4.4.1.1 Post operative complications 65 4.4.1.2 Chemotherapy complications 65 4.5 Results of parameters studied 69 4.5.1 Physiological parameters 69 4.5.2 Hematology 69 4.5.2.1 Total leukocyte count 69 4.5.2.2 Haemoglobin 71 4.5.2.3 Packed cell volume 71 4.5.2.4 Differential leukocyte count 73 4.5.2.4.1 Neutrophils 73 4.5.2.4.2 Lymphocytes 76 4.5.2.4.3 Eosinophils 78 4.5.2.4.4 Monocytes 78 4.5.3 Biochemical observations 80 4.5.3.1 Calcium (mg%) 82 4.5.3.2 Phosphorus (mg%) 82 4.5.3.3 Blood urea nitrogen (mg%) 85 4.5.3.4 Creatinine (mg%) 87 4.6 Pathological observations 87 4.6.1 Gross pathological observations 87 4.6.2 Histopathology 89 4.6.2.1 Anal tumors 91 4.6.2.2 Perineal tumors 95 4.7 AgNOR count 98 4.8 Immunohistochemistry 100 4.9 Recurrence, disease free intervals and prognosis 100 V DISCUSSION 102 VI SUMMARY 121 LITERATURE CITED 124 vii LIST OF ILLUSTRATIONS Table Page TITLE No. No. 1. Showing vaginal examination in a dog using index finger 37 2. Showing the topography of the perineal region in a bitch 37 3. Animal with perineal swelling and passing ribbon like stools 37 4. Gross examination of tumor growth 37 5. Fine needle aspiration biopsy collection from a mass in a dog 38 6. Collecting impression smears from the growth in a dog 38 7. Showing the position of a dog with ventral recumbency with elevated hind 41 Portion 8. Showing the mass which was explored via sharp and blunt dissection 41 9. Showing the skin sutures of the surgical excision with the anal canal patency 41 maintenance 10. Showing auto analyser PRIETEST for biochemical analysis and kits 44 11. Pie chart depicting the age wise incidence of anal and perineal tumors 51 12. Pie chart showing the breed wise incidence of anal and perineal tumors 53 13. Sexwise incidence of anal and perineal tumors 55 14. Showing hepatoid tumor with severe ulceration 57 15. Showing blood discharge from anus in a dog affected with hepatoid 57 adenocarcinoma 16. Showing tail lifting in a dog affected with anal tumor 57 17. Showing the lethargy dog affected with anal tumor 57 18. Showing the necrosed mass in a dog affected with perineal tumor 57 19. Note small circumscribed, ulcerated nodules on the perianal tissue in hepatoid 59 adenocarcinoma 20. Note erythematous, ulcerated, lemon sized mass of anal sac adenocarcinoma 59 in a dog 21. Note cauliflower like, multilobulated, red colored mass in TVT 59 22. Note the freely movable mass of basal cell carcinoma in a dog 59 23. Showing cauliflower like, hard mass of squamous cell carcinoma in a dog 59 24. Photograph showing myxofibrosarcoma with extensive ulceration in a dog 59 viii 25. Note the bunches of grapes appearance in myxosarcoma 61 26. Anal sac adenocarcinoma: Note pleomorphic, homogenous, clusters of 61 cuboidal cells with several nucleoli. H& E x 700 27. Anal sac adenocarcinoma: Note anisocytosis and anisokaryosis with 61 vacuolated cytoplasm H& E x 700 28. Hepatoid adenocarcinoma: Note polygonal or clusters of cells showing 61 anisokaryosis having several nucleoli. H& E x 700 29. TVT: Note several lymphocytes and plasma cells with cytoplasmic 61 vacuolation. H& E x 700 30. TVT: Note the extensive infiltration of neutrophils. H& E x 700 61 31. Basal cell carcinoma: Note aggregation of small, uniform cells with basophilic 62 cytoplasm in groups. H& E x 700 32. Squamous cell carcinoma: Cytological smear showing large squamous epithelial cells with prominent irregular nuclei and neutrophilic infiltration. 62 H& E x 700 33. Lateral radiograph showing the radiolucent areas in the ribs in animals with 63 hepatoid adenocarcinoma 34. Lateral radiograph showing soft tissue density in the perineal region 63 35. Showing the anal sac adenocarcinoma in a dog 64 36. Showing the anus plugged with mop to avoid contamination 64 37. Showing the incision over the mass 64 38. Showing the excision of the mass 64 39. Showing the sutured skin after excision of the mass 64 40. Showing the recurrence of anal sac adenocarcinoma in a dog 66 41. Photograph showing cystic papillary adenocarcinoma with extensive 66 ulceration in a dog before giving chemotherapy 42. Photograph showing the reduced size of lesion after 3 weeks of chemotherapy 66 43. Showing squamous cell carcinoma with pedunculated mass in urogenital 67 perineal area 44. Showing the manual separation of mass from the vagina 67 45. Showing the ligation of the vaginal pedicle 67 46. Showing pedunculated growth of squamous cell carcinoma 67 47. Showing the perineal area after surgery 67 48. Showing the TVT on vulva before chemotherapy 68 ix 49. Showing the reduced lesion of TVT after 2 weeks of chemotherapy 68 50. Showing fistulation at the anal region and loss of anal sphincter in a dog 68 51. Showing extravasation of chemotherapy in animals with anal tumors 68 52. Cut section of cystic papillary adenocarcinoma with abundant brownish black 90 cystic fluid 53. Cut section of myxosarcoma soft, grey to white slimy mass 90 54 (a). Graph showing histological types of anal tumors in dogs 92 54 (b). Graph showing histological types of perineal tumors in dogs 92 55. Anal gland tumor and Hepatoid gland tumor: Note proliferation of glandular epithelial cells and hepatoid nature of the cell proliferation with connective 94 tissue stroma. H& E x 280 56. Myxofibrosarcoma: Section showing plump proliferated fibroblast and 94 stellate cells with processes. H& E x 70 57. Hepatoid adenocarcinoma: Note the islands of tumor cells supported by 94 connective tissue stroma. H& E x 70 58. Hepatoid adenocarcinoma: Note the pleomorphic hepatoid cells with 94 acidophilic cytoplasm. H& E x 280 59. Hepatoid adenocarcinoma: Section showing pleomorphic hepatoid cells with 94 area of congestion. H& E x 280 60. Basal cell carcinoma: Section showing proliferated basal cells in dermis in the 94 form of islands supported by connective tissue stroma. H& E x 70 61. Basal cell carcinoma: Note proliferated basal cells in dermis. H& E x 280 96 62. Cystic papillary adenocarcinoma: Note irregular acinar structures with cystic 96 dilatation and infiltration of neutrophils. H& E x 280 63. Cystic papillary adenocarcinoma: Note dilated glandular structures and proliferated cells giving papillary projection and infiltration of neutrophils. 96 H& E x 280 64. TVT: Note sheets of round to individual cells separated by fine connective 96 tissue stroma. H& E x 280 65. Squamous cell carcinoma: Note the irregular masses of epithelial cells and 96 keratin pearls. H& E x 70 66. Squamous cell carcinoma: Section showing keratin pearls with concentric 96 layers of squamous cells. H& E x 280 67. Myxosarcoma: Note stellate calls arranged in whirl pattern with processes. 97 H& E x 70 68. Myxosarcoma: Note pleomorphic stellate cells with processes lying in 97 basophilic mucinous matrix. H& E x 280 x
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