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Clinico-haematobiochemical studies and surgico-chemotherapeutic management of canine dermatological neoplasms.

Introduction

Canine skin is most prone for tumors. Skin tumors are more likely to be benign than malignant (Wilcock, 1993). Of all species, dog develops neoplasm twice as frequently as humans, with incidence of skin and mammary tumours being the highest (Nair et al., 2007).

Invention of advanced techniques has helped in prevention and control of infectious diseases resulting in more dogs living in cancer age (Krithiga et al., 2005). Bronden et al. (2009) concluded that treatment of skin neoplasms, in particular malignant tumours, can be challenging. The treatment of choice in most cases includes surgical excision but depends upon the type of cancer, stage, grade and location. Ettinger et al. (2003) advised that role of chemotherapy is most appropriate in treatment of connective tissue tumour and mainly used to treat incompletely resected tumours, high grade tumours and metastatic tumour.

The present investigation is aimed to provide data on clinico-haematobiochemical changes of skin and subcutaneous tumours and their surgicochemotherapeutic management in canines.

Materials and Methods

A total of 750 cases presented during October' 2010 to September' 2011 and out of these 40 dogs were affected with skin and subcutaneous neoplastic growths at various body parts. Twenty five (25) clinical cases of skin and subcutaneous tumors in dogs of different age groups and breeds were selected for present study. Clinical parameters of animals like general condition, visual examination of tumour mass, consistency, location of tumour and their size/weight were recorded. Blood samples were collected before surgery and at 60, 120 and 180 minutes during and post-surgical intervention. Briefly, following human handling, approximately 3 ml blood was collected by vein puncture either from cephalic or saphenous vein in previously marked vials containing ethylene diamine tetra acetic acid (EDTA) and blood smears were made on clean marked glass slides using a drop of blood without anticoagulant for DLC. The estimation of hematological parameters viz. Haemoglobin (Hb), Packed cell volume (PCV), Total leucocyte count (TLC) were done by automated haematology blood cell counter (Sussi, France) and Differential leucocyte count (DLC) was performed following Leishman's staining method.

For estimation of biochemical parameters viz. Blood Glucose, Total proteins, Blood urea nitrogen (BUN), Creatinine, Aspartate aminotransferase (AST) and Alanine aminotransferase (ALT) serum samples were harvested by routine manner. Serum samples were collected before surgery, post-surgery and after application of antineoplastic drug i.e. Vincristine sulphate and Methotrexate. These parameters were estimated by standard methods using Semi-Automated Analyzer (Logotech-Techo-168).

The data obtained were subjected to statistical analysis as per the procedure described by Snedecor and Cochran (1994).

For surgical interventions, the animals were divided into three groups i.e. group I, fifteen dogs which were affected with benign tumours were treated surgically. In group II, five dogs which were affected with malignant tumours were treated surgically while remaining five dogs of group III which were affected with malignant tumours were treated by lumpectomy plus antineoplastic therapy (i.e. Vincristine sulphate @ 0.025 mg/kg b. wt. I/V followed one week later by Methotrexate @ 0.3 mg/kg b. wt.) intravenously at weekly interval under fluid therapy. All the dogs of Group I, II and III were pre-medicated with Atropine sulphate @ 0.65 mg (total dose) i/m followed by Xylazine sedation @ 1mg/kg. b. wt. or Medetomidine @ 20 [micro]g/kg, i/m and general anaesthesia was achieved by using Ketamine @ 5 mg/ kg i/m or Propofol @ 5 mg/kg. b. wt. i/v.

Post-operatively, Dextrose saline (200ml) was given to ensure rehydration and renal function. Injection of Intacef (a) 500mg for five days, Melonex (a) 2 ml for three days was given intramuscularly. Supportive therapy with both chemotherapeutic agents the hepatoprotectant, Tribiveta @ 0.05-0.2 mg/kg b.wt. I/M was given for 5 days followed by Polybion (b) syrup 1 tsf bid and Tab Liv-52 (c) every day during course of chemotherapy. Post operatively, wound was dressed with Povidone iodine ointment and its spray for eight days. The skin sutures were removed on tenth postoperative day.

Results and Discussion Clinical parameters

In the present study, 80% of reported clinical cases were in good body condition and recovered uneventfully except cases of high grade malignancy. The occurrence of non pedunculated (broad based) and tumours in hard consistency were higher accounting for 72% and 60% respectively (Fig. 1-4). The oral and venereal tumours were soft in consistency. The size/weight of tumour mass ranged from 0 to 300 grams with a frequency of 44% (0-50 gms), 28% (50-100 gms), 20% (100-200 gms) and 8% (200-300 gms) respectively (Fig.5-8). However, Gupta and Tiwari (2009) reported that weight ranged from 20 gms to a maximum of 1.25kg. The results of our study are in close agreement with earlier reports (Palta, 2000; Mason, 2007; Vani et al., 2007). The reason for delayed healing might be due to large surgical wound, poor health condition, decreased immunity, improper management by owners and post-operative infections.

Haematological parameters

The results of haematology have been summarized and presented in Table 1. Briefly, haemoglobin (g/dl), PCV (%) and lymphocytes (%) registered non-significant decrease, however TLC (thousands/cu.mm) and neutrophil (%) showed a non significant increase among various groups. Further, the proportions of monocytes (%), basophils (%) and eosinophils (%) showed non-significant fluctuation.

The results of haematology corroborates with the reports by Pavesi et al. (1995) and Stockhaus et al. (1999). Riley and Riley (1982) reported that neutropenia, thrombocytopenia, fall in total WBC count and PCV generally occurred following treatment with Vincristine, Methotrexate and Cyclophosphamide in dogs. Vegad (2007) also found that values of haemoglobin starts increasing post-surgery which might be due to haemo-concentration of red cells because of tissue alteration in acute inflammation and increased viscosity of blood. The value started decreasing after chemotherapy which might be due to chemotherapeutic damage to red blood cell formation in bone marrow or due to haemorrhage (Martineau, 2006). Pre-operative TLC values were high due to leucomoied reaction (Vegad, 2007) but after surgery the value started decreasing. Ravikumar et al. (1999) also found a sharp decrease in lymphocyte count immediately after surgery. The decrease in haematological values might be due to bone marrow suppression. Antineoplastic therapy induced myelo-suppression as cytotoxic drugs suppress replicating precursor cells of bone marrow and attributed to significant haematological changes (Jajleen, 2010).

Biochemical parameters

The results of blood biochemistry have been summarized and presented in Table 2 and 3. Briefly, a significant (p < 0.05) elevation in activities of ALT and AST was recorded. Similarly, levels of blood glucose and serum urea nitrogen also increased significant (p < 0.05). Moreover, total protein and creatinine showed nonsignificant decrease among groups.

The increase in blood glucose level in present study could be attributed to increased adrenocortical hormones during anaesthesia as a result of stress, mobilization of liver glycogen under influence of increased adrenaline level, decreased glucose utilization or impaired insulin activity and antineoplastic drug induced stress (Lee Femine et al., 1957). Bisen et al. (1994) also reported a significant increase in blood glucose between 1 to 2 hours after Ketamine administration. Alike our results, Benjamin (1985) also reported significant decrease in total protein in response to neoplastic growth involving lymphoid tissue and liver.

SUN and creatinine are the most common tests to assess renal function in dogs. The rise in creatinine along with increase in SUN might be due to temporary inihibitory effect of Ketamine on renal blood flow and use of antineoplastic therapy as also reported by Palta (2000) and Nayyar (2002) in female dogs suffering from mammary tumour. Mostly chemotherapeutic drugs are nephrotoxic in nature due to which there may be sudden rise in value of serum creatinine.

The rise in AST might be attributed to hepatic insufficiency in old patients affected with mammary tumour or skin and subcutaneous tumours. Further, it might also be due to alteration in cell membrane permeability due to antineoplastic drugs. Parallel finding have been reported by Brar and Sandhu (2000) in dogs affected with mammary tumour. There was significant rise in ALT values after Vincristine and Methotrexate administration which might be because of mild hepatotoxic effect of drugs but it was under physiological limit and in agreement with Abudu (2009).

Surgico-Chemotherapeutic Management

The dogs were followed up at least for 2 months after surgery. Four dogs from group II and five dogs from group III did not show any recurrences. One dog of group II showed recurrence in 4th month post-surgery and dog died one month after recurrence of tumor. Lumpectomy plus Vincristine sulphate and Methotrexate therapy was found to be the most successful therapy for treatment of skin and subcutaneous tumours. Although side effects like alopecia, anorexia, vomition and anaemia were observed with Vincristine sulphate and Methotrexate.

Our results of surgico-chemotherapeutic management are analogous to earlier reports of Ettinger et al. (2003); Nak et al. (2005); Enhert (2008); Khan et al. (2009); Pakhrin et al. (2009) and Sharda et al. (2011). Alike our findings Bronden et al. (2009) and Bhatia et al. (2010) also advocated that treatment of malignant neoplasms can be challenging. The treatment of choice in most cases includes surgical excision but depends upon the type of cancer, stage, grade and location. In some cases radiation or chemotherapy may be used alone or as adjunctive therapy for malignant neoplasms.

Based on these observations it can be concluded that there is no significant effect of skin and subcutaneous neoplasms on physiological parameters whereas variations seen in clinical parameters. Alterations in hematological and biochemical values, probably depend upon immune status, hormonal changes, diet and the relative number in different geographical areas. In the present study surgical excision followed by chemotherapy was found most effective treatment for canine skin and subcutaneous neoplasia. Similar findings have also been reported by Johnson (2005) and Pandey (2000) for the treatment of neoplasms in canines.

References

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Brar, R.S. and Sandhu, H.S. (2000). Veterinary Clinical Diagnosis by Laboratory Methods. 1st Edn. Kalyani Publisher, New Delhi, pp: 28-131.

Bisen, S.S., Pandey, S.K., Sharma, I.J. Chandrapuria, V.P.and Bhargava, M.K. (1994). Biochemical and haematological effects of certain analgesic premedicants with ketamine hydrochloride in dogs. Ind. J. Vet. Surg. 15: 30-31.

Bronden, Louise B., Eriksen, Thomas. and T Kristensen, Annemarie. (2009). Oral malignant melanomas and other head and neck neoplasms in Danish dogs -data from the Danish Veterinary Cancer Registry. Act. Vet. Scand. 51: 54.

Bhatia, Ami, Tank, P.H., Kavechiya, V.P., Vedpathak, H.S., and Karle, A.S. (2010). Clinical Management of Canine Transmissible Venereal Granuloma in Dogs. The Ind. J. Field. Vet. 4: 73-75.

Ettinger, Stephen J., Feldman. and Edward, C. (2003). Textbook of Veterinary Internal Medicine (4th ed.). W.B. Saunders Company.

Enrhart, N. (2008). Soft tissue sarcomas in dogs. J. Amer.Vet. Med. Asso. 41: 241-46.

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Jajleen, P.W. (2010). About Low White Blood Cell Counts. http://www.fda.gov/medwatch/chemotherapy. com/chemotherapy.com.

Krithiga, K., Murali Manohar, B. and Balchandran, C. (2005). Cytological and histopathological diagnosis of canine skin and adnexae cell tumours. Ind. J. Vet. Pathol. 29: 112-17.

Khan, L.A., Khante, G.S., Raut, B.M., Bodkhe, A.M., Chavan, M.S., Pagrut, N.S. and Bobde, S.P. (2009). Incidence of Venereal Granuloma and its Medicinal treatment in stray Dogs of Nagpur. City. Vet World. 2: 13-14.

Lee Feminae, A., Marks, J.L., Teter, J.C., Fattin, J.H.L., Leonard, M.P. and Braker, D.V. (1957). The adrenocortical response in surgical patient. Anim. Surg.146: 26.

Martineau, C.M. (2006). Efficacy, Cost, Toxicity and Duration of treatment by Chemotherapy. Vet. Clin. North Amer. (S. Anim. Pract.). 26: 63-71.

Mason, L. K. (2007). Treatment of contaminated wounds.In: Harari, J.(ed), Saunders Company, Philadelphia. pp 33-60.

Nak, D., Nak, Y, Cangul, I.T and Tuna, B (2005). A clinico-pathological study on the effect of vincristine on transmissible venereal tumours in the dog. Ind. J. Vet. Pathol. 52: 366-70.

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Nayyar, A. (2002). Evaluation of vinblastine and mitomycin in conjugation with surgery in canine mammary neoplasms. Post GraduateThesis, PAU, Ludhiana, India.

Pavesi, L., Preti, P.G., Da Prada Pedrazzoli, P., Poggi, G. and Robustelli della, C.G. (1995). Epirubicin versus mitoxantrone in combination Chemotherapy for metastatic breast cancer. Anticancer Res. 15: 495-501.

Palta, M.K. (2000). Clinical studies on multimodality in the management of canine mammary neoplasm. Post GraduateThesis, PAU, Ludhiana, India.

Pakhrin, Bidur., Kang, Min-Soo., Bae, Il-Hong., Park, Mi-Sun., Jee, Hyang., You, Mi-Hyeon., Kim, Jae-Hoon., Yoon, Byung-Il., Choi, Yang-Kyu. and Kim, Dae-Yong. (2009). Retrospective study of canine cutaneous tumours. Kor. J. Vet. Sci. 8: 229-36.

Pandey, S.K. (2000). Management of recurring squamous cell carcinoma of palate with combined therapy. Ind. J. Vet. Surg. 21: 57-58.

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Ravikumar, K.P., Vasanth, M.S. and Jayadevappa, S.M. (1999). Chemotherapeutic trial on mammary tumours in canines. Ind. J. Vet. Surg. 20: 115.

Snedecor, G.W. and Cochran, W.G. (1994). In: Statistical Methods, 8th edition, East West Press Pvt. Ltd., New Delhi.

Sharda, R., Tiwari, S.K., Nath, K., Naik, S. and Gurmita, K.K. (2011). Management of fibrosarcoma of upper palate and nasal bones in a dog. Ind. Vet. J. 88: 60-61.

Stockhaus, C., Kohn, B., Rudolph, R., Brunnberg, L. and Giger, U. (1999). Correlation of haemostatic abnormalities with tumour stage and characteristics in dogs with mammary carcinoma. J. Smal. Anim. Pract. 40: 326-31.

Vani, G., Haragopal, V., Suresh Kumar, R.V., Rrilatha, Ch. and Rao, T.S. Chandrasekhar (2007). Oral Tumours in Canines. Ind. Vet. J. 84: 1083-1085.

Vegad, J. (2007). A Textbook of General Veterinary Pathology. 1st Edn., Vikas Publishing House, New Delhi., pp 204-288.

Wilcock, B. P. (1993). Skin and appendages. In Jubb, K. F. Pathology of domestic animals. Academic press. INC, California. pp 706-33.

Deepak Kumar Kashyap (1), S.K. Tiwari, D. K. Giri, Govina Dewangan and B. Sinha

Department of Veterinary Surgery and Radiology

College of Veterinary Science and Animal Husbandry

Chhattisgarh Kamdhenu University

Anjora

Durg--491001 (Chhattisgarh)

* Part of Post Graduate thesis submitted

(1.) Corresponding author: E-mail:deepakkashyap31@gmail.com

(a)--Brand of Intas Animal Health, Ahmedabad

(b)--Brand of Merck Limited, Mumbai

(c) -Brand of Himalaya Healthcare, Bengaluru

Table 1: Haematological observations before and after
treatment in dogs affected with skin and subcutaneous
neoplasms.

Parameters          Groups   Post operative time interval
                    (n=15    (min.)
                    in I         0 min.          60 min.
                    and
                    n=5 in
                    II,
                    III)

Hb (gm/dl)            I      13.65 [+ or -]   12.28 [+ or -]
                               0.2 (a)          0.24 (b)
                      II     11.12 [+ or -]   11.48 [+ or -]
                               0.40 (b)         0.23 (b)
                     III     11.96 [+ or -]   12.84 [+ or -]
                               0.23 (b)         0.15 (a)
PCV (%)               I      35.98 [+ or -]   36.92 [+ or -]
                               1.66 (a)         1.78 (b)
                      II     36.88 [+ or -]   33.84 [+ or -]
                               1.48 (b)         1.80 (c)
                     III     33.80 [+ or -]   35.15 [+ or -]
                               1.39 (b)         1.46 (a)
TLC (x [10.sup.3]     I      17.20 [+ or -]   12.51 [+ or -]
                               0.13 (b)         0.09 (a) **
  cu [mm.sup.-1])     II     17.81 [+ or -]   13.81 [+ or -]
                               0.12 (a)         0.08 (a) **
                     III     16.41 [+ or -]   12.81 [+ or -]
                               0.12 (a)       0.08 (a) **
Neutrophils (%)       I      64.67 [+ or -]   64.93 [+ or -]
                               0.71 (a)         0.75 (a)
                      II     70.00 [+ or -]   70.80 [+ or -]
                               0.63 (b)         0.97 (b)
                     III     68.20 [+ or -]   68.60 [+ or -]
                               0.37 (b)         1.44 (b)
Lymphocytes (%)       I      30.73 [+ or -]   30.47 [+ or -]
                               0.53 (a)         0.62 (a)
                      II     25.40 [+ or -]   24.60 [+ or -]
                             0.24 (b)           0.98 (b)
                     III     26.20 [+ or -]   25.80 [+ or -]
                             0.49 (b)           1.24 (b)

Parameters          Groups   Post operative time interval
                    (n=15    (min.)
                    in I        120 min.         180 min.
                    and
                    n=5 in
                    II,
                    III)

Hb (gm/dl)            I      11.16 [+ or -]   12.92 [+ or -]
                               0.34 (c)         0.22 (a)
                      II     12.43 [+ or -]   10.12 [+ or -]
                               0.18 (a)         0.83 (b)
                     III     11.72 [+ or -]   10.28 [+ or -]
                               0.25 (b)         0.22 (b)
PCV (%)               I      33.88 [+ or -]   35.62 [+ or -]
                               1.10 (c)         1.59 (a)
                      II     35.57 [+ or -]   36.84 [+ or -]
                               1.52 (a)         1.39 (b)
                     III     36.90 [+ or -]   33.50 [+ or -]
                               1.84 (b)         1.65 (b)
TLC (x [10.sup.3]     I      9.61 [+ or -]    9.23 [+ or -]
                               0.08 (b)  **     0.12 (a) **
  cu [mm.sup.-1])     II     9.91 [+ or -]    9.20 [+ or -]
                               0.08 (a) **      0.16 (a) **
                     III     9.91 [+ or -]    9.20 [+ or -]
                               0.06 (a) **      0.16 (a) **
Neutrophils (%)       I      66.07 [+ or -]   65.0 [+ or -]
                               0.82 (a)         0.45 (a)
                      II     71.0 [+ or -]    70.2 [+ or -]
                               0.71 (a)         1.05 (a)
                     III     66.80 [+ or -]   66.4 [+ or -]
                               1.02 (a)         0.81 (c)
Lymphocytes (%)       I      30.13 [+ or -]   30.50 [+ or -]
                               0.86 (a)         0.43 (b)
                      II     23.80 [+ or -]   24.80 [+ or -]
                               0.49 (a)         1.08 (c)
                     III     25.40 [+ or -]   26.00 [+ or -]
                               1.33 (a)         0.58 (a)

** P<0.01= Significant at 1% level

Table 2 : Biochemical observations before and after surgical
treatment in dogs affected with skin and subcutaneous
neoplasms.

Parameters            Groups (n=15         0 mins.
                      in I and
                      n=5 in II)

Glucose                    I         78.00 [+ or -] 1.18
(mg/dl)                    II        80.60 [+ or -] 1.24
Total Proteins             I          6.4 [+ or -] 0.141
(gm/dl)                    II         5.46 [+ or -] 0.14
Serum Urea Nitrogen        I         17.13 [+ or -] 0.41
(mg/dl)                    II        18.20 [+ or -] 1.15
Creatinine                 I          1.20 [+ or -] 0.13
(mg/dl)                    II         1.15 [+ or -] 0.24
ALT                        I         27.80 [+ or -] 1.092
(U/L)                      II        29.40 [+ or -] 0.748
AST                        I         48.73 [+ or -] 1.40
(U/L)                      II        47.20 [+ or -] 1.62

Parameters            Groups (n=15   Post surgery (mins.)
                      in I and
                      n=5 in II)

Glucose                    I         85.73 [+ or -] 0.95 *
(mg/dl)                    II        88.60 [+ or -] 1.72 *
Total Proteins             I         5.02 [+ or -] 0.07 *
(gm/dl)                    II        4.62 [+ or -] 0.08 *
Serum Urea Nitrogen        I         21.53 [+ or -] 0.78 *
(mg/dl)                    II         21.20 [+ or -] 1.28
Creatinine                 I          0.85 [+ or -] 0.02
(mg/dl)                    II         0.96 [+ or -] 0.02
ALT                        I          38.13 [+ or -] 0.63
(U/L)                      II        42.40 [+ or -] 1.93 *
AST                        I         55.40 [+ or -] 1.21 *
(U/L)                      II         54.80 [+ or -] 1.49

* P<0.05= Significant at 5% level

Table 3 : Biochemical observations before, after surgery and
post antineoplastic treatment in dogs affected with skin and
subcutaneous neoplasms.

Parameters   Groups   0 mins.
             (n=5)

Glucose       III     73.60 [+ or -] 1.02 (c)
  (mg/dl)
Total         III     5.54 [+ or -] 0.20 (a)
  Proteins
  (gm/dl)
Serum Urea    III     21.80 [+ or -] 1.28 (a)
  Nitrogen
  (mg/dl)
Creatinine    III     1.28 [+ or -] 0.05 (a)
  (mg/dl)
ALT (U/L)     III     35.60 [+ or -] 1.029 (c)
AST (U/L)     III     46.40 [+ or -] 0.67 (c)

Parameters   Post surgery and
             Antineoplastic therapy
             Post         Post          Post
             operative    Vincristine   Methotrexate
             (mins.)      (mins.)       (mins.)

Glucose      79.20        77.40         84.20
  (mg/dl)      [+ or -]     [+ or -]      [+ or -]
               2.31         2.50          3.20
               (b) *        (b) *         (a) *
Total        5.22         4.60          4.68
  Proteins     [+ or -]     [+ or -]      [+ or -]
  (gm/dl)      0.07 (a)     0.20          0.124
                            (b) *         (b) *
Serum Urea   24.00        25.80         24.40
  Nitrogen     [+ or -]     [+ or -]      [+ or -]
  (mg/dl)      0.70         1.06          1.43
               (a) *        (a) *         (a) *
Creatinine   1.20         1.32          2.80
  (mg/dl)      [+ or -]     [+ or -]      [+ or -]
               0.05 (a)     1.54 (a)      0.04 (a)
ALT (U/L)    42.00        51.60         50.20
               [+ or -]     [+ or -]      [+ or -]
               1.22         3.35          1.06
               (b) *        (a) **        (a) **
AST (U/L)    51.60        57.80         59.80
               [+ or -]     [+ or -]      [+ or -]
               1.86         1.79          1.59
               (b) *        (a) **        (a) **

** P<0.01= Significant at 1% level
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Title Annotation:Research Article
Author:Kumar Kashyap, Deepak; Tiwari, S.K.; Giri, D.K.; Dewangan, Govina; Sinha, B.
Publication:Intas Polivet
Article Type:Report
Geographic Code:9INDI
Date:Jan 1, 2014
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